土三七诱导小鼠HVOD模型的建立及丹参对HVOD预防性治疗的实验研究
|
摘要
研究背景：肝小静脉闭塞病(hepatic veno-occlusive disease, HVOD)又称肝窦阻塞综合征(hepatic sinusoidal obstruction syndrome,HSOS),晦床表现为肝大、腹水、体重增加和黄疸。常因骨髓移植前大剂量使用细胞毒药物和免疫抑制剂预处理如环磷酰胺、白消胺等和／或全身放疗及服用含吡咯烷生物碱药物引起。肝移植、肺移植术后亦可发生HVOD。HVOD是骨髓移植后(HSCT)的三大并发症之一，发生率达10%-50%,重度死亡率达90%以上。预防和治疗HVOD是移植患者预后良好的关键。我国栽种土三七面广,土三七服用人群多。近年来国内因服土三七预防老年心血管疾患致肝小静脉闭塞病的报道明显增多。目前HOVD发生机制尚不明确,建立HVOD模型是研究其发生机制的关键,国内外实验室已投入大量人力物力进行研究。迄今为止关于HVOD模型已有多种,但各有利弊。Borowska等以亚硝酸胺建立HVOD模型,但病变程度一般较轻,较适合轻、中度HVOD研究。Deleve等及陈妙妍等利用大剂量野百合碱对大鼠进行灌胃,建立急性HVOD模型,成模率较高,但个体差异大,病变程度不一,造模周期短,预防用药疗效观察时间短,远期疗效不确切。国内有学者利用土三七生物碱提取成分建立HVOD模型,成模率低,并且未对该模型进行评价。目前对HVOD治疗措施有限,各种药物的疗效存在很大争议。实验室已经发现一些药物(如tPA、低分子肝素、前列腺素E1、激素、N-乙酰半胱氨酸、去纤苷等)在预防和治疗HVOD有一定作用,但其确切疗效仍存在争议。在国际上有关HVOD药物的研发领域中,目前主要是针对HVOD预防药物的研究。例如,熊去氧胆酸(优思弗)对早期HVOD有效,对中晚期患者疗效欠佳,用药途径主要是口服用药,对于伴有恶心、呕吐或口腔疾病的大部分中晚期患者限制了药物的应用。强的松、低分子肝素等对早期HVOD有一定作用,但副作用大,易出现大出血等严重并发症,临床应用需十分慎重。去纤苷是目前国际上认为治疗HVOD较有效的药物,但国内应用较少。丹参是一种改善循环的药物,其作用表现在改善心、肝、肺、脑等脏器的缺血再灌注损伤；研究发现丹参作用还表现在对肝细胞的损伤、肝纤维化、肝硬变、肝癌的治疗及调节免疫应答、抗感染和抗肿瘤等方面。丹参中的丹参素对血小板、白细胞和血管内皮细胞表达的各种细胞黏附分子具有抑制作用和抗血栓形成的作用。临床治疗中发现丹参对预防移植术后HVOD有效,丹参对土三七诱导的HVOD是否有预防作用?丹参通过什么途径对HVOD起作用?是本课题旨在研究的重要问题。本实验通过环磷酰胺、野百合碱作为模型对照,并与云南三七比较,利用土三七建立HVOD模型,并对该模型进行：①小鼠一般情况观察；②肝功能损害检测；③病理学评分。同时,本实验利用不同剂量的丹参对土三七模型进行干预,研究血管粘附分子相关改变,初步探讨丹参对HVOD防治效果及其防治作用机制。
目的：建立土三七诱导的肝小静脉闭塞病模型。利用磷酸缓冲液、云南三七作为正常对照组,用环磷酰胺、野百合碱、土三七三种药物诱导肝小静脉闭塞病(HVOD)模型,比较各模型的利弊,为HVOD的研究提供良好的造模方法。
方法：将110只雌性昆明小鼠随机分为五组,分别予磷酸缓冲液(PBS)、云南三七、环磷酰胺、野百合碱(MCT)、土三七灌胃,并分别在灌胃后30天、30天、30天、7天、30天处死小鼠,测肝重／体重、ALT、AST、ALB、TBIL;对肝组织进行HE染色、Masson染色,并用修改后Deleve评分标准进行病理学评分。
结果：
1.PBS组和云南三七组无小鼠成模,环磷酰胺组、野百合碱组、土三七组分别有5只、18只、24只成模。
2.各模型组与PBS对照组比较,小鼠肝重／体重、血清ALT、AST、TBIL明显升高,ALB明显降低,差异有统计学意义(P<0.05)；各模型组间肝重／体重、肝功能无显著差异(P>0.05)。
3.根据修改后Deleve评分标准,各模型组病理学改变符合HVOD,模型组评分土三七组、环磷酰胺组成模小鼠大部分表现为中、重度HVOD,野百合碱组成模小鼠大部分表现为轻、中度。
结论：环磷酰胺、土三七、野百合碱均能诱导HVOD模型,土三七和野百合碱组成模率较环磷酰胺高。土三七诱导的HVOD模型符合人类HVOD病理改变,为HVOD进一步研究提供了新的造模方法。
目的：观察丹参对土三七诱导小鼠HVOD的预防治疗作用。
方法：将100只雌性昆明小鼠随机分为四组,分别为磷酸缓冲液(PBS)组、土三七组、丹参预防治疗组(分两个剂量组：100mg/kg, 200mg/kg)。各组分别连续灌胃30天,灌胃结束后处死小鼠,留取血清、肝组织。测肝重／体重、ALT、AST、ALB、TBIL;对肝组织进行HE染色、Masson染色,并用修改后Deleve评分标准进行病理学评分。用免疫组化法和RT-PCR法检测肝组织PAI-1和VEGF表达水平
结果：
1.土三七组、丹参100mg/kg预防治疗组、丹参200mg/kg预防治疗组分别有24只、8只、3只小鼠发生HVOD。
2.丹参预防治疗组与土三七模型组比较,小鼠肝重／体重、小鼠血清ALT、AST、TBIL明显降低,ALB明显升高,差异有统计学意义(P<0.05)；丹参200mg/kg预防治疗组与丹参100mg/kg预防治疗组比较效果好,丹参200mg/kg预防治疗组小鼠肝重／体重明显减小,小鼠血清ALT、AST、TBIL明显降低,血清ALB明显升高,差异有统计学意义(P<0.05)。
3.各组小鼠肝脏病理评分结果：土三七组发生HVOD小鼠肝脏病理评分大部分表现以中、重度为主；丹参100mg/kg预防治疗组HVOD小鼠多表现为轻、中度,仅1例为重度；丹参200mg/kg预防治疗组发生HVOD小鼠多为轻度,无一例表现为重度。
4.肝组织PAI-1及VEGF免疫组化结果：丹参预防治疗组与土三七模型组比较,PAI-1和VEGF水平显著降低(P<0.05),丹参200mg/kg预防治疗组PAI-1与VEGF水平比丹参100mg/kg预防治疗组下降更明显(P<0.05)。
5.肝组织RT-PCR检测PAI-1及VEGF的mRNA结果：丹参预防治疗组与土三七模型组比较,PAI-1和VEGF mRNA水平显著降低(P<0.05),丹参200mg/kg预防治疗组PAI-1和VEGF mRNA水平比丹参100mg/kg预防治疗组显著降低(P<0.05)。
结论：
1.丹参预防治疗土三七诱导HVOD有效小鼠的血清ALT、AST、TBIL下降、ALB增加,减轻肝窦内皮细胞和肝细胞病理损害,从而降低肝组织病理学评分。
2.不同剂量丹参可减少土三七诱导小鼠HVOD的发生。丹参组肝脏PAI-1和VEGF水平明显降低,HVOD发生率低,丹参200mg/kg组较丹参100mg/kg组预防治疗效果显著。丹参组小鼠肝脏PAI-1和VEGF水平降低,病情程度相对较轻,提示HVOD小鼠预后较好,肝脏PAI-1和VEGF表达水平检测可作为HVOD病情程度及预后监测指标。
目的：观察丹参对土三七诱导HVOD小鼠肝脏肿瘤坏死因子a、血管内皮生长因子、血管细胞间粘附分子1、细胞间粘附分子1、核转录因子p65 mRNA和蛋白水平及血管内皮生长因子受体1、血管内皮生长因子受体2的mRNA水平,探讨丹参预防HVOD的作用机制。
方法：(1)每组随机取4只小鼠肝脏行RT-PCR检测：肿瘤坏死因子a、血管内皮生长因子、血管内皮生长因子受体1、血管内皮生长因子受体2、血管细胞间粘附分子1、细胞间粘附分子1、核转录因子mRNA表达。(2)每组随机取5只小鼠肝脏行western bloting检测肝组织肿瘤坏死因子a、血管内皮生长因子、血管细胞间粘附分子l、细胞间粘附分子1、核转录因子p65的蛋白水平。
结果：
1.肝组织RT-PCR检测的mRNA结果：肿瘤坏死因子a、血管内皮生长因子、血管内皮生长因子受体1、血管内皮生长因子受体2、血管细胞间粘附分子1、细胞间粘附分子1、核转录因子p65 mRNA表达模型组比PBS组显著升高(P<0.05)；丹参预防治疗组比模型组显著降低(P<0.05)。
2.肝组织western bloting检测蛋白结果：肝组织肿瘤坏死因子a、血管内皮生长因子、血管细胞间粘附分子1、细胞间粘附分子1、核转录因子p65蛋白表达模型组比PBS组显著升高(P<0.05)；丹参预防治疗组比模型组显著降低(P<0.05)。
结论：
1.丹参可能通过下调肝组织中VCAM-1、ICAM-1、NF-κBp65 mRNA和蛋白表达,抑制炎性因子的表达,减少渗出,抑制血栓形成,减少HVOD发生。
2.丹参可能通过下调肝组织中VEGF mRNA和蛋白表达及VEGF-R1、VEGF-R2 mRNA的表达,阻止肝窦内皮细胞的迁移,减少内皮通透性,从而减少HVOD发生。
丹参是一种活血化瘀的中成药,已有报道丹参对预防心脑血管血栓形成具有重要作用。临床发现丹参对移植术后HVOD发生具有显著效果。丹参对土三七诱导的HVOD有无作用?通过什么途径起作用是本课题研究需解决的重要问题。
本研究首次运用土三七煎液灌胃诱导小鼠HVOD模型,并对该模型进行病理学评分。用丹参预防治疗土三七诱导的小鼠HVOD模型来观察丹参预防性治疗HVOD的效果,并初步探讨丹参预防治疗HVOD发生的作用机制。本研究采用血清生化学,病理学的HE染色、Masson染色、免疫组织化学染色,RT-PCR、Western bloting等方法,从血清生化、组织、mRNA、蛋白的水平来评价丹参预防性治疗HVOD的效果和探讨丹参预防HVOD发生的作用机制。结果：①土三七能诱导小鼠HVOD发生,成模率高,病理学改变符合人类HVOD变化。②丹参具有降低小鼠血清ALT、AST、TBIL,增加ALB,减少肝窦淤血,减轻肝窦内皮细胞和肝细胞变性、坏死的作用。③丹参具有降低小鼠肝脏PAI-1、VEGF mRNA和蛋白表达作用,PAI-1、VEGF可作为评价HVOD预后标志之一。④丹参具有下调小鼠肝组织中TNF-a、VCAM-1、ICAM-1、NF-κBp65 mRNA和蛋白表达,抑制炎性因子的表达,防止血栓形成,减少HVOD发生的作用。⑤丹参具有下调肝组织VEGF mRNA和蛋白表达及VEGF-R1、VEGF-R2 mRNA的表达的作用。结论：土三七能成功诱导小鼠HVOD发生。丹参对土三七诱导的小鼠HVOD有预防性治疗作用。其作用机制可能与减少炎症因子,降低肝组织VCAM-1、ICAM-1、NF-κBP65 mRNA和蛋白表达,预防血栓形成有关。丹参可降低VEGF mRNA和蛋白表达及VEGF-R1、VEGF-R2 mRNA的表达,从而阻止肝窦内皮细胞的迁移、坏死,减少内皮细胞间通透性,预防肝窦淤血。本课题研究结果为丹参临床治疗HVOD提供实验依据。
Backgroud:Hepatic veno-occlusive(HVOD) another name is hepatic sinusoidal obstruction sydrome(HSOS).HVOD is characterized by tender hepatomegaly,fluid retention,weight gain,and jaundice.This condition is seen after hematopoietic stem cell transplantation(HSCT),high dose abdominal radiation therapy, use of certain chemotherapeutic agents,ingestion of pyrrolizidine alkaloids.Liver transplantation and lung transplantation can also lead to HVOD. HVOD is one of the three complications after HSCT.The incidence of HVOD varies from 10% to 50%, and the motality more than 90%.Preventing and treating HVOD plantation patients ealier are the key points for eusemia. Gynura segetum is one kind of Chinese materia medica which extensively planted. The reports about HVOD induced by Gynura segetum is increasing rencently years.Mechanisms for HVOD have not been identified, so the model for HVOD is the key target for research and internal and abroad have thrown much to it. Varies models have establised with advantages and disadvantages. Borowska et al induced HVOD model by nitrous acid amine which is not ideal because of the low fraction of animals with typical lesions,long period and high variability.Monocrotalinethe model establised by Deleve et al and Chen MY have made great progress in model cycle and achievement ratio,but they were limited to observe prostecdtive effiacy. Chinese researches have established mouse HVOD model by alkaloids of Sedum aizoon, but without evaluation. The treatments for HVOD are limited, and the drugs for HVOD are disputing. Some drugs effective for HVOD have been found to some extent (such as tPA, Low Molecular Heparin, prostaglandin E1, prednisone,N-acetyl cysteine,defibrotide et al). Nowadays,the reserarch about HVOD is popular with the prophylactic agent in international.For example,Ursode-oxycholic acid is good for early HVOD,but useless for late HVOD,and the administration route by oral is always limited for late HVOD patients with nausea,vomito or oral disease. Prednisone, low molecular heparin have some effect to early HVOD,but the side effect of hemorrhea is more graveness. Defibrotide is one kinds of effective drugs,but haven't taken to clinic extensivly in China. Salvia miltiorrhiza is one kind of drug good for circulation,especially for injury cardia,liver, lung and brain ischemia-reperfusion injury. It also found that Salvia miltiorrhiza is used for hepatocellular injury,hepatic fibrosis,hepatic cirrhosis, liver cancer, immune response of rugulation, anti-infection and anti-tumor. Tanshinol, the ingredient of salvia miltiorrhiza, can inhibit the cellular adhesion molecule express by platelet, leucocyte and endotheliocyte and the thromb. Clinic researches have found that Salvia miltiorrhiza have effect on preventing HVOD after transplantation.However, its preventive and therapeutic effects on HVOD induced by gynura segetum remain unclear. Constructing a mice model of HVOD induced by gynura segetum and establishing the control models by cyclophosphamide,Yunnan sanqi,mon-ocrotaline.The present study is to investigate the anti-HVOD effects of Salvia miltiorrhiza, the changes of adhesion molecule and the related molecular mechanisms.
Objective:To establish the hepatic veno-occlusive disease reduced by gynura segetum. Phosphate buffer and Yunnan Sanqi as the nomal control groups. Comparing the mice models of Hepatic Veno-occlusive disease induced by cyclophosphamide, monocrotaline and gynura segetum to help advance research into this disease.
Methods:110 female KunMing mice were randomly classified into five groups. PBS, Yunnan Sanqi, cyclophosphamide, monocrotaline or gynura segetum was administered. They were sacrificed on day 30,30,30,7,30. The weight of the liver and body were measured; blood samples were collected to determine alanine aminotransferase (ALT), aspartate aminotransferase (AST),albumin (ALB), total bilirubin(TBIL). Liver were sectioned and stained using HE and Masson to observe the pathological changes, and the changes on light microscopy were assessed by a modified Deleve scoring system.
Result:The group of PBS and Yunnan Sanqi have no mouse with HVOD.5 of cyclophosphamide group,18 of monocrotaline group and 24 mice of gynura segetum group presented with clinical symptoms and the histopathological picture of HVOD. To compare with PBS group, the model groups had significant changes in the level of serum ALT, AST, ALB, TBIL and the weight of liver and body(P<0.05), and there were no obvious changes between the model groups. The model groups of liver had obvious pathological changes of HVOD. Most of gynura segetum and cyclophosphamide group with HVOD were in middle or late phases, while monocrotaline group were in early or middle phases.
Conclusion:gynura segetum, monocrotaline and cyclophosphamide could induce the model of HVOD successfully. The model induced by gynura segetum and monocrotaline were higher than cyclophosphamide. The model induced by gynura segetum exhibited the clinical and histogical features of human HVOD, and this model may provide a new model experimental approach for the further study of the human HVOD.
Objective:To investigate the preventive treatment effects of Salvia miltiorrhiza on mice hepatic veno-occlusive induced by gynura segetum.
Methods:100 female KunMing mice were randomly classified into five groups. The control group were treated with PBS, model group induced by gynura segetum, gynura segetum groups preventively treated with Salvia miltiorrhiza at two dosages(gynura segetum+Salvia miltiorrhiza,100mg/kg,200mg/kg).The mice were sacrificed on day 30. The weight of the liver and body were measured; blood samples were collected to determine alanine aminotransferase (ALT), aspartate aminotransferase (AST),albumin (ALB), total bilirubin(TBIL). Liver were sectioned and stained using HE and Masson to observe the pathological changes, and the changes on light microscopy were assessed by a modified Deleve scoring system. The protein expression of PAI-1 and VEGF in mice liver were detected by immunohistochemical staining and the mRNA expression by RT-PCR.
Result:24 mice of gynura segetum group,8 of 100mg/kg Salvia miltiorrhiza group and 3 of 200mg/kg Salvia miltiorrhiza group presented with clinical symptoms and the histopathological picture of HVOD. To compare with gynura segetum group, the Salvia miltiorrhiza groups had significant decreased in the level of serum ALT, AST, ALB, TBIL and the weight of liver and body(P<0.05), and there were also significant changes between the Salvia miltiorrhiza groups(P<0.05). most of gynura segetum and cyclophosphamide group with HVOD were in middle or late phases, while 100mg/kg Salvia miltiorrhiza group were in early or middle phases and only one mouse presented with late HVOD. Most of 200mg/kg Salvia miltiorrhiza group mice with HVOD were presented early and no one was severe. The immunohistochemical staining demonsrated that the protein expression levels of PAI-1 and VEGF were singnificantly lower in 100mg/kg Salvia miltiorrhiza group and 200mg/kg Salvia miltiorrhiza group than that in gynura segetum group(P<0.05,respectively).The mRNA expression detection results by RT-PCR were consistent with that of protein expression levels measured by immunohistochemical staining(P<0.05,respectively). Compared with 100mg/kg Salvia miltiorr-hiza group, PAI-1 and VEGF of protein and mRNA expression in 200mg/kgSalvia miltiorrhiza group had decreased significantly(P<0.05, respectively).
Conclusion:Salvia miltiorrhiza had significant biological function of sinusoidals and hepatocyte protection indicated by decreasing serum ALT,AST and TBIL levels,increasing serum ALB level, attenuating the modified Deleve scores of mice liver tissue induced by gynura segetum. Salvia miltiorrhiza with different dosages had therapeutic effect on HVOD induced by gynura segetum, and the Salvia miltiorrhiza 200mg/kg group did better than Salvia miltiorrhiza 100mg/kg group. The decrease of PAI-1 and VEGF protein and mRNA expression indicated better prognosis of HVOD, and detecting the expression of PAI-1 and VEGF may be used as monitors of the prognosis of HVOD.
Objective:To explore the molecular mechanisms of the preventive treatment of salvia miltiorrhiza on mice HVOD induced by gynura segetum through the investigation of mRNA and protein expression of tumor necrosis factor-α(TNF-α), vascular endothelial growth factor(VEGF), vascular cellular adhesion molecule-1(VC AM-1), intercellular adhesion molecule-1(ICAM-1),nuclear trascription factor P65(NF-κBp65) and the mRNA expression of vascular endothelial growth factor recptor-1(flt-1), vascular endothelial growth factor recptor-2(KDR) in mice liver tissue.
Methods:The liver tissue specimens were randomly collected from four mice in each group for RT-PCR and five mice in each group for western bloting.RT-PCR was applied to detect the mRNA expression of TNF-α, VCAM-1, ICAM-1, VEGF, flt-1, KDR,NF-KBp65.Western blot was employed to measure the protein expression of TNF-α, VCAM-1, ICAM-1, VEGF, NF-KBp65 in liver tissue.
Result.The RT-PCR demonstrated that the mRNA expression levels of TNF-α, VCAM-1, ICAM-1, VEGF, flt,KDR,and NF-κBp65 were singnificantly lower in salvia miltiorrhiza groups than that in gynura segetum group (P<0.05,respectively).The protein expression detection results by western bloting were consist with that of mRNA expression levels by RT-PCR(P<0.05,respectively).
Conclusion:Salvia miltiorrhiza may attenuate mice HVOD via down-regulating mRNA and protein expression of TNF-α，VCAM-1, ICAM-1 and NF-κBp65 and inhibiting the expression of inflammatory factor and thrombogenesis.Meanwhile, salvia miltiorrhiza may prevent the HOVD by inhibiting the migration and necrosis of sinusoidal endothielial cell through down-regulating the mRNA and protein expression VEGF and the mRNA expression of flt、KDR in liver tissue.
Salvia miltiorrhiza is one kind of chinese patent medicine.they have effect on promoting blood flow and preventing the cardial and cerebral vascular thrombosis.Clinic researches demonstrated that salvia miltiorrhiza had significant effect on preventing HVOD after transplantation. However, wether it has preventive and therapeutic effects on HVOD induced by gynura segetum and by what way remain unclear
Gynura segetum juice was applied for the first time to Construct a mice model of HVOD in the present study and to assess the model by pathological assay. Salvia miltiorrhiza was first applied for preventively treat HVOD mice model and the mechanism was analyzed. The preventive treatment effects of salvia miltiorrhiza were assessed by both pathological and serological assays,such as HE tissue staining, Masson collagen staining.The mechanisms were explored respectively at histology, mRNA, RT-PCR and Western bloting.The results demonstrated that mice HVOD can be induced by gynura segetum and the pathological changes was similar to human HVOD. Salvia miltiorrhiza significantly decreased mice serum ALT,AST and TBIL,elevated serum ALB level, decreased sinusoidal endothielial cell necrosis and sinusoidal congestion, and inhibited the degeneration and necrosis of hepatic cell. Salvia miltiorrhiza can down-regulated the mRNA and protein expression of PAI-1 and VEGF,which had significant point to assess the prognosis of HVOD. Meanwhile, salvia miltiorrhiza attenuated mice HVOD via down-regulating mRNA and protein expression of TNF-a, VCAM-1, ICAM-1 and NF-κBp65 and inhibiting the expression of inflammatory factor and thrombogenesis. What's more, salvia miltiorrhiza also down-regulated the protein expression of VEGF, and the mRNA expression of VEGF, flt and KDR.
The present study demonstrates the gynura segetum can induced mice HVOD and the preventative treatment effects of salvia miltiorrhiza on mice HVOD induced by gynura segetum. The molecular mechanisms were supposed to be associated with the attenuation of liver inflammatory factor,the inhibition on the expression of VCAM-1, ICAM-1, NF-KBp65, VEGF, VEGF-R1 and VEGF-R2,as well as prevention of thrombogenesis and migration and necrosis of sinusoidal endothielial cell.
目的：建立土三七诱导的肝小静脉闭塞病模型。利用磷酸缓冲液、云南三七作为正常对照组,用环磷酰胺、野百合碱、土三七三种药物诱导肝小静脉闭塞病(HVOD)模型,比较各模型的利弊,为HVOD的研究提供良好的造模方法。
方法：将110只雌性昆明小鼠随机分为五组,分别予磷酸缓冲液(PBS)、云南三七、环磷酰胺、野百合碱(MCT)、土三七灌胃,并分别在灌胃后30天、30天、30天、7天、30天处死小鼠,测肝重／体重、ALT、AST、ALB、TBIL;对肝组织进行HE染色、Masson染色,并用修改后Deleve评分标准进行病理学评分。
结果：
1.PBS组和云南三七组无小鼠成模,环磷酰胺组、野百合碱组、土三七组分别有5只、18只、24只成模。
2.各模型组与PBS对照组比较,小鼠肝重／体重、血清ALT、AST、TBIL明显升高,ALB明显降低,差异有统计学意义(P<0.05)；各模型组间肝重／体重、肝功能无显著差异(P>0.05)。
3.根据修改后Deleve评分标准,各模型组病理学改变符合HVOD,模型组评分土三七组、环磷酰胺组成模小鼠大部分表现为中、重度HVOD,野百合碱组成模小鼠大部分表现为轻、中度。
结论：环磷酰胺、土三七、野百合碱均能诱导HVOD模型,土三七和野百合碱组成模率较环磷酰胺高。土三七诱导的HVOD模型符合人类HVOD病理改变,为HVOD进一步研究提供了新的造模方法。
目的：观察丹参对土三七诱导小鼠HVOD的预防治疗作用。
方法：将100只雌性昆明小鼠随机分为四组,分别为磷酸缓冲液(PBS)组、土三七组、丹参预防治疗组(分两个剂量组：100mg/kg, 200mg/kg)。各组分别连续灌胃30天,灌胃结束后处死小鼠,留取血清、肝组织。测肝重／体重、ALT、AST、ALB、TBIL;对肝组织进行HE染色、Masson染色,并用修改后Deleve评分标准进行病理学评分。用免疫组化法和RT-PCR法检测肝组织PAI-1和VEGF表达水平
结果：
1.土三七组、丹参100mg/kg预防治疗组、丹参200mg/kg预防治疗组分别有24只、8只、3只小鼠发生HVOD。
2.丹参预防治疗组与土三七模型组比较,小鼠肝重／体重、小鼠血清ALT、AST、TBIL明显降低,ALB明显升高,差异有统计学意义(P<0.05)；丹参200mg/kg预防治疗组与丹参100mg/kg预防治疗组比较效果好,丹参200mg/kg预防治疗组小鼠肝重／体重明显减小,小鼠血清ALT、AST、TBIL明显降低,血清ALB明显升高,差异有统计学意义(P<0.05)。
3.各组小鼠肝脏病理评分结果：土三七组发生HVOD小鼠肝脏病理评分大部分表现以中、重度为主；丹参100mg/kg预防治疗组HVOD小鼠多表现为轻、中度,仅1例为重度；丹参200mg/kg预防治疗组发生HVOD小鼠多为轻度,无一例表现为重度。
4.肝组织PAI-1及VEGF免疫组化结果：丹参预防治疗组与土三七模型组比较,PAI-1和VEGF水平显著降低(P<0.05),丹参200mg/kg预防治疗组PAI-1与VEGF水平比丹参100mg/kg预防治疗组下降更明显(P<0.05)。
5.肝组织RT-PCR检测PAI-1及VEGF的mRNA结果：丹参预防治疗组与土三七模型组比较,PAI-1和VEGF mRNA水平显著降低(P<0.05),丹参200mg/kg预防治疗组PAI-1和VEGF mRNA水平比丹参100mg/kg预防治疗组显著降低(P<0.05)。
结论：
1.丹参预防治疗土三七诱导HVOD有效小鼠的血清ALT、AST、TBIL下降、ALB增加,减轻肝窦内皮细胞和肝细胞病理损害,从而降低肝组织病理学评分。
2.不同剂量丹参可减少土三七诱导小鼠HVOD的发生。丹参组肝脏PAI-1和VEGF水平明显降低,HVOD发生率低,丹参200mg/kg组较丹参100mg/kg组预防治疗效果显著。丹参组小鼠肝脏PAI-1和VEGF水平降低,病情程度相对较轻,提示HVOD小鼠预后较好,肝脏PAI-1和VEGF表达水平检测可作为HVOD病情程度及预后监测指标。
目的：观察丹参对土三七诱导HVOD小鼠肝脏肿瘤坏死因子a、血管内皮生长因子、血管细胞间粘附分子1、细胞间粘附分子1、核转录因子p65 mRNA和蛋白水平及血管内皮生长因子受体1、血管内皮生长因子受体2的mRNA水平,探讨丹参预防HVOD的作用机制。
方法：(1)每组随机取4只小鼠肝脏行RT-PCR检测：肿瘤坏死因子a、血管内皮生长因子、血管内皮生长因子受体1、血管内皮生长因子受体2、血管细胞间粘附分子1、细胞间粘附分子1、核转录因子mRNA表达。(2)每组随机取5只小鼠肝脏行western bloting检测肝组织肿瘤坏死因子a、血管内皮生长因子、血管细胞间粘附分子l、细胞间粘附分子1、核转录因子p65的蛋白水平。
结果：
1.肝组织RT-PCR检测的mRNA结果：肿瘤坏死因子a、血管内皮生长因子、血管内皮生长因子受体1、血管内皮生长因子受体2、血管细胞间粘附分子1、细胞间粘附分子1、核转录因子p65 mRNA表达模型组比PBS组显著升高(P<0.05)；丹参预防治疗组比模型组显著降低(P<0.05)。
2.肝组织western bloting检测蛋白结果：肝组织肿瘤坏死因子a、血管内皮生长因子、血管细胞间粘附分子1、细胞间粘附分子1、核转录因子p65蛋白表达模型组比PBS组显著升高(P<0.05)；丹参预防治疗组比模型组显著降低(P<0.05)。
结论：
1.丹参可能通过下调肝组织中VCAM-1、ICAM-1、NF-κBp65 mRNA和蛋白表达,抑制炎性因子的表达,减少渗出,抑制血栓形成,减少HVOD发生。
2.丹参可能通过下调肝组织中VEGF mRNA和蛋白表达及VEGF-R1、VEGF-R2 mRNA的表达,阻止肝窦内皮细胞的迁移,减少内皮通透性,从而减少HVOD发生。
丹参是一种活血化瘀的中成药,已有报道丹参对预防心脑血管血栓形成具有重要作用。临床发现丹参对移植术后HVOD发生具有显著效果。丹参对土三七诱导的HVOD有无作用?通过什么途径起作用是本课题研究需解决的重要问题。
本研究首次运用土三七煎液灌胃诱导小鼠HVOD模型,并对该模型进行病理学评分。用丹参预防治疗土三七诱导的小鼠HVOD模型来观察丹参预防性治疗HVOD的效果,并初步探讨丹参预防治疗HVOD发生的作用机制。本研究采用血清生化学,病理学的HE染色、Masson染色、免疫组织化学染色,RT-PCR、Western bloting等方法,从血清生化、组织、mRNA、蛋白的水平来评价丹参预防性治疗HVOD的效果和探讨丹参预防HVOD发生的作用机制。结果：①土三七能诱导小鼠HVOD发生,成模率高,病理学改变符合人类HVOD变化。②丹参具有降低小鼠血清ALT、AST、TBIL,增加ALB,减少肝窦淤血,减轻肝窦内皮细胞和肝细胞变性、坏死的作用。③丹参具有降低小鼠肝脏PAI-1、VEGF mRNA和蛋白表达作用,PAI-1、VEGF可作为评价HVOD预后标志之一。④丹参具有下调小鼠肝组织中TNF-a、VCAM-1、ICAM-1、NF-κBp65 mRNA和蛋白表达,抑制炎性因子的表达,防止血栓形成,减少HVOD发生的作用。⑤丹参具有下调肝组织VEGF mRNA和蛋白表达及VEGF-R1、VEGF-R2 mRNA的表达的作用。结论：土三七能成功诱导小鼠HVOD发生。丹参对土三七诱导的小鼠HVOD有预防性治疗作用。其作用机制可能与减少炎症因子,降低肝组织VCAM-1、ICAM-1、NF-κBP65 mRNA和蛋白表达,预防血栓形成有关。丹参可降低VEGF mRNA和蛋白表达及VEGF-R1、VEGF-R2 mRNA的表达,从而阻止肝窦内皮细胞的迁移、坏死,减少内皮细胞间通透性,预防肝窦淤血。本课题研究结果为丹参临床治疗HVOD提供实验依据。
Backgroud:Hepatic veno-occlusive(HVOD) another name is hepatic sinusoidal obstruction sydrome(HSOS).HVOD is characterized by tender hepatomegaly,fluid retention,weight gain,and jaundice.This condition is seen after hematopoietic stem cell transplantation(HSCT),high dose abdominal radiation therapy, use of certain chemotherapeutic agents,ingestion of pyrrolizidine alkaloids.Liver transplantation and lung transplantation can also lead to HVOD. HVOD is one of the three complications after HSCT.The incidence of HVOD varies from 10% to 50%, and the motality more than 90%.Preventing and treating HVOD plantation patients ealier are the key points for eusemia. Gynura segetum is one kind of Chinese materia medica which extensively planted. The reports about HVOD induced by Gynura segetum is increasing rencently years.Mechanisms for HVOD have not been identified, so the model for HVOD is the key target for research and internal and abroad have thrown much to it. Varies models have establised with advantages and disadvantages. Borowska et al induced HVOD model by nitrous acid amine which is not ideal because of the low fraction of animals with typical lesions,long period and high variability.Monocrotalinethe model establised by Deleve et al and Chen MY have made great progress in model cycle and achievement ratio,but they were limited to observe prostecdtive effiacy. Chinese researches have established mouse HVOD model by alkaloids of Sedum aizoon, but without evaluation. The treatments for HVOD are limited, and the drugs for HVOD are disputing. Some drugs effective for HVOD have been found to some extent (such as tPA, Low Molecular Heparin, prostaglandin E1, prednisone,N-acetyl cysteine,defibrotide et al). Nowadays,the reserarch about HVOD is popular with the prophylactic agent in international.For example,Ursode-oxycholic acid is good for early HVOD,but useless for late HVOD,and the administration route by oral is always limited for late HVOD patients with nausea,vomito or oral disease. Prednisone, low molecular heparin have some effect to early HVOD,but the side effect of hemorrhea is more graveness. Defibrotide is one kinds of effective drugs,but haven't taken to clinic extensivly in China. Salvia miltiorrhiza is one kind of drug good for circulation,especially for injury cardia,liver, lung and brain ischemia-reperfusion injury. It also found that Salvia miltiorrhiza is used for hepatocellular injury,hepatic fibrosis,hepatic cirrhosis, liver cancer, immune response of rugulation, anti-infection and anti-tumor. Tanshinol, the ingredient of salvia miltiorrhiza, can inhibit the cellular adhesion molecule express by platelet, leucocyte and endotheliocyte and the thromb. Clinic researches have found that Salvia miltiorrhiza have effect on preventing HVOD after transplantation.However, its preventive and therapeutic effects on HVOD induced by gynura segetum remain unclear. Constructing a mice model of HVOD induced by gynura segetum and establishing the control models by cyclophosphamide,Yunnan sanqi,mon-ocrotaline.The present study is to investigate the anti-HVOD effects of Salvia miltiorrhiza, the changes of adhesion molecule and the related molecular mechanisms.
Objective:To establish the hepatic veno-occlusive disease reduced by gynura segetum. Phosphate buffer and Yunnan Sanqi as the nomal control groups. Comparing the mice models of Hepatic Veno-occlusive disease induced by cyclophosphamide, monocrotaline and gynura segetum to help advance research into this disease.
Methods:110 female KunMing mice were randomly classified into five groups. PBS, Yunnan Sanqi, cyclophosphamide, monocrotaline or gynura segetum was administered. They were sacrificed on day 30,30,30,7,30. The weight of the liver and body were measured; blood samples were collected to determine alanine aminotransferase (ALT), aspartate aminotransferase (AST),albumin (ALB), total bilirubin(TBIL). Liver were sectioned and stained using HE and Masson to observe the pathological changes, and the changes on light microscopy were assessed by a modified Deleve scoring system.
Result:The group of PBS and Yunnan Sanqi have no mouse with HVOD.5 of cyclophosphamide group,18 of monocrotaline group and 24 mice of gynura segetum group presented with clinical symptoms and the histopathological picture of HVOD. To compare with PBS group, the model groups had significant changes in the level of serum ALT, AST, ALB, TBIL and the weight of liver and body(P<0.05), and there were no obvious changes between the model groups. The model groups of liver had obvious pathological changes of HVOD. Most of gynura segetum and cyclophosphamide group with HVOD were in middle or late phases, while monocrotaline group were in early or middle phases.
Conclusion:gynura segetum, monocrotaline and cyclophosphamide could induce the model of HVOD successfully. The model induced by gynura segetum and monocrotaline were higher than cyclophosphamide. The model induced by gynura segetum exhibited the clinical and histogical features of human HVOD, and this model may provide a new model experimental approach for the further study of the human HVOD.
Objective:To investigate the preventive treatment effects of Salvia miltiorrhiza on mice hepatic veno-occlusive induced by gynura segetum.
Methods:100 female KunMing mice were randomly classified into five groups. The control group were treated with PBS, model group induced by gynura segetum, gynura segetum groups preventively treated with Salvia miltiorrhiza at two dosages(gynura segetum+Salvia miltiorrhiza,100mg/kg,200mg/kg).The mice were sacrificed on day 30. The weight of the liver and body were measured; blood samples were collected to determine alanine aminotransferase (ALT), aspartate aminotransferase (AST),albumin (ALB), total bilirubin(TBIL). Liver were sectioned and stained using HE and Masson to observe the pathological changes, and the changes on light microscopy were assessed by a modified Deleve scoring system. The protein expression of PAI-1 and VEGF in mice liver were detected by immunohistochemical staining and the mRNA expression by RT-PCR.
Result:24 mice of gynura segetum group,8 of 100mg/kg Salvia miltiorrhiza group and 3 of 200mg/kg Salvia miltiorrhiza group presented with clinical symptoms and the histopathological picture of HVOD. To compare with gynura segetum group, the Salvia miltiorrhiza groups had significant decreased in the level of serum ALT, AST, ALB, TBIL and the weight of liver and body(P<0.05), and there were also significant changes between the Salvia miltiorrhiza groups(P<0.05). most of gynura segetum and cyclophosphamide group with HVOD were in middle or late phases, while 100mg/kg Salvia miltiorrhiza group were in early or middle phases and only one mouse presented with late HVOD. Most of 200mg/kg Salvia miltiorrhiza group mice with HVOD were presented early and no one was severe. The immunohistochemical staining demonsrated that the protein expression levels of PAI-1 and VEGF were singnificantly lower in 100mg/kg Salvia miltiorrhiza group and 200mg/kg Salvia miltiorrhiza group than that in gynura segetum group(P<0.05,respectively).The mRNA expression detection results by RT-PCR were consistent with that of protein expression levels measured by immunohistochemical staining(P<0.05,respectively). Compared with 100mg/kg Salvia miltiorr-hiza group, PAI-1 and VEGF of protein and mRNA expression in 200mg/kgSalvia miltiorrhiza group had decreased significantly(P<0.05, respectively).
Conclusion:Salvia miltiorrhiza had significant biological function of sinusoidals and hepatocyte protection indicated by decreasing serum ALT,AST and TBIL levels,increasing serum ALB level, attenuating the modified Deleve scores of mice liver tissue induced by gynura segetum. Salvia miltiorrhiza with different dosages had therapeutic effect on HVOD induced by gynura segetum, and the Salvia miltiorrhiza 200mg/kg group did better than Salvia miltiorrhiza 100mg/kg group. The decrease of PAI-1 and VEGF protein and mRNA expression indicated better prognosis of HVOD, and detecting the expression of PAI-1 and VEGF may be used as monitors of the prognosis of HVOD.
Objective:To explore the molecular mechanisms of the preventive treatment of salvia miltiorrhiza on mice HVOD induced by gynura segetum through the investigation of mRNA and protein expression of tumor necrosis factor-α(TNF-α), vascular endothelial growth factor(VEGF), vascular cellular adhesion molecule-1(VC AM-1), intercellular adhesion molecule-1(ICAM-1),nuclear trascription factor P65(NF-κBp65) and the mRNA expression of vascular endothelial growth factor recptor-1(flt-1), vascular endothelial growth factor recptor-2(KDR) in mice liver tissue.
Methods:The liver tissue specimens were randomly collected from four mice in each group for RT-PCR and five mice in each group for western bloting.RT-PCR was applied to detect the mRNA expression of TNF-α, VCAM-1, ICAM-1, VEGF, flt-1, KDR,NF-KBp65.Western blot was employed to measure the protein expression of TNF-α, VCAM-1, ICAM-1, VEGF, NF-KBp65 in liver tissue.
Result.The RT-PCR demonstrated that the mRNA expression levels of TNF-α, VCAM-1, ICAM-1, VEGF, flt,KDR,and NF-κBp65 were singnificantly lower in salvia miltiorrhiza groups than that in gynura segetum group (P<0.05,respectively).The protein expression detection results by western bloting were consist with that of mRNA expression levels by RT-PCR(P<0.05,respectively).
Conclusion:Salvia miltiorrhiza may attenuate mice HVOD via down-regulating mRNA and protein expression of TNF-α，VCAM-1, ICAM-1 and NF-κBp65 and inhibiting the expression of inflammatory factor and thrombogenesis.Meanwhile, salvia miltiorrhiza may prevent the HOVD by inhibiting the migration and necrosis of sinusoidal endothielial cell through down-regulating the mRNA and protein expression VEGF and the mRNA expression of flt、KDR in liver tissue.
Salvia miltiorrhiza is one kind of chinese patent medicine.they have effect on promoting blood flow and preventing the cardial and cerebral vascular thrombosis.Clinic researches demonstrated that salvia miltiorrhiza had significant effect on preventing HVOD after transplantation. However, wether it has preventive and therapeutic effects on HVOD induced by gynura segetum and by what way remain unclear
Gynura segetum juice was applied for the first time to Construct a mice model of HVOD in the present study and to assess the model by pathological assay. Salvia miltiorrhiza was first applied for preventively treat HVOD mice model and the mechanism was analyzed. The preventive treatment effects of salvia miltiorrhiza were assessed by both pathological and serological assays,such as HE tissue staining, Masson collagen staining.The mechanisms were explored respectively at histology, mRNA, RT-PCR and Western bloting.The results demonstrated that mice HVOD can be induced by gynura segetum and the pathological changes was similar to human HVOD. Salvia miltiorrhiza significantly decreased mice serum ALT,AST and TBIL,elevated serum ALB level, decreased sinusoidal endothielial cell necrosis and sinusoidal congestion, and inhibited the degeneration and necrosis of hepatic cell. Salvia miltiorrhiza can down-regulated the mRNA and protein expression of PAI-1 and VEGF,which had significant point to assess the prognosis of HVOD. Meanwhile, salvia miltiorrhiza attenuated mice HVOD via down-regulating mRNA and protein expression of TNF-a, VCAM-1, ICAM-1 and NF-κBp65 and inhibiting the expression of inflammatory factor and thrombogenesis. What's more, salvia miltiorrhiza also down-regulated the protein expression of VEGF, and the mRNA expression of VEGF, flt and KDR.
The present study demonstrates the gynura segetum can induced mice HVOD and the preventative treatment effects of salvia miltiorrhiza on mice HVOD induced by gynura segetum. The molecular mechanisms were supposed to be associated with the attenuation of liver inflammatory factor,the inhibition on the expression of VCAM-1, ICAM-1, NF-KBp65, VEGF, VEGF-R1 and VEGF-R2,as well as prevention of thrombogenesis and migration and necrosis of sinusoidal endothielial cell.
引文
[1]DeLeve LD, Shulman HM, McDonald GBToxic injury to hepatic sinusoids:sinusoidal obstruction syndrome (veno-occlusive disease). Semin Liver Dis.2002,22:27-42
[2]Brice, K., B. Valerie, et al. Risk-adjusted monitoring of veno-occlusive disease following Bayesian individualization of busulfan dosage for bone marrow transplantation in paediatrics. Pharmacoepidemiol Drug Saf.2008,17(2):135-143.
[3]Cacchione, A., A. LeMaitre, et al.. "Risk factors for hepatic veno-occlusive disease:a retrospective unicentric study in 116 children auto grafted after a high-dose BU-thiotepa regimen." Bone Marrow Transplant.2008,42(7):449-454.
[4]Carreras, E., L. Rosinol, et al. "Veno-occlusive disease of the liver after high-dose cytoreductive therapy with busulfan and melphalan for autologous blood stem cell transplantation in multiple myeloma patients." Biol Blood Marrow Transplant.2007,13(12):1448-1454.
[5]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and The rapeutics. 2006,23(1),11-25.
[6]Membreno, F. E., J. Ortiz, et al. Liver transplantation for sinusoidal obstructive syndrome (veno-occlusive disease):case report with review of the literature and the UNOS database. Clin Transplant.2008,22(4):397-404.
[7]de Fontbrune, F. S., H. Mal, et al. Veno-occlusive disease of the liver after lung transplantation. Am J Transplant.2007,7(9):2208-2211.
[8]Du, H., D.-L. Yang, et al. Hepatic veno-occlusive disease after hematopoietic stem cell transplantation in 23 patients. Zhonghua Gan Zang Bing Za Zhi.2007,15(12):949-950.
[9]Chen, M.-Y., J.-T. Cai, et al. Hepatic veno-occlusive disease associated with the use of Gynura segetum. Eur J Intern Med.2007,18(8):609.
[10]Wu, X.-j., K. Zhang, et al. Clinical analysis of four cases of hepatic
veno-occlusive disease caused by Gynura segetum (Lour.) Merr.Zhonghua Gan Zang Bing Za Zhi.2007,15(2):151-153.
[11]BorowskaH,AieiszewskaE,CzaudemaP,et,al.Daetinomycin-induced Veno-occlus-ive disease in rats. The hepatoprotective action of amifostine. Evaluation in a light and electron microscope.Folia Morph-ologica.2004,63:91-93
[12]Deleve LD et al. Characterization of Reproducible Rat Model of Hepatic Veno-occlusive Disease.Hepatology.1999,29:1779-1791.
[13]Chen MY, Cai JT et al. Reliable experimental model of hepatic veno-occlusive disease cause by monocrotaline [J].Hepatobiliary Pancreat Dis Int.2008,7(4):395-400.
[14]Gao XS, Xiao SH,et al. Analysis of alkaloids in sedum aizoon and establishement of hepatic veno-occlusive model in mice. Chin J Integr Trad West Med Dig.2006,14:311-313.
[15]Bajwa, r.P., cant, A.J., Abinun, m., Flood, T.J., Hodges, s., Hale, J.P., et al. recombinant tissue plasminogen activator for treatment of hepatic veno-occlusive disease following bone marrow transplantation in children:Effectiveness and a scoring system for initiating treatment. Bone Marrow Transplantation.2003,31(7), 591-597.
[16]litzow, m.r., repoussis, P.D., schroeder, g., schembri-Wismayer, D., Batts, K.P., Anderson, P.m, et al..veno-occlusive disease of the liver after blood and marrow transplantation:Analysis of pre-and post-transplant risk factors associated with severity and results of therapy with tissue plasminogen activator. Leukemia and Lymphoma.2002,43(11),2099-2107.
[17]Batsis, i., yannaki, E., Kaloyannidis, P., sakellari, i., smias, c., georgoulis, i., et al. veno-occlusive disease prophylaxis with fresh frozen plasma and heparin in bone marrow transplantation. Thrombosis Research.2006,118(5),611-618.
[18]Imran, H., Tleyjeh, i.m., Zirakzadeh, A., rodriguez, v.,& Khan, s.P.use of prophylactic anticoagulation and the risk of hepatic veno-occlusive disease in patients undergoing hematopoietic stem cell transplantation:A systematic review and meta-analysis. Bone Marrow Transplantation.2006,37(7),677-686.
[19]Sun, X., W.-G. Hao, et al. Efficacy of low-dose heparin and prostaglandin El in the prevention of hepatic veno-occlusive disease after allogenic hematopoietic stem cell transplantation in children with beta-thalassemia major. Zhongguo Dang Dai Er Ke Za Zhi.200,9(4):343-346.
[20]Al Beihany, A., H. Al Omar, et al. Successful treatment of hepatic veno-occlusive disease after myeloablative allogeneic hematopoietic stem cell transplantation by early administration of a short course of methylprednisolone. Bone Marrow Transplant.2008,41(3):287-291.
[21]Ringden, O., remberger, m., lehmann, s., Hentschke, P., mattsson, J., Klaesson, s., et al. N-acetylcysteine for hepatic veno-occlusive disease after allogeneic stem cell transplantation. Bone Marrow Transplant.2000,25(9),993-996.
[22]Bairey, O., Kirgner, i., yakobi, m., Hamdan, A., Ben-Ari, Z.,& shaklai, m. clinical severe hepatic veno-occlusive disease during induction treatment of acute monoblastic leukemia managed with deibrotide. American Journal of Hematology.2002,69(4),281-284.
[23]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics. 2006,23(1),11-25.
[24]Dignan, F., gujral, D., Ethell, m., Evans, s., Treleaven, J., morgan, g., et al. Prophylactic deibrotide in allogeneic stem cell transplantation:minimal morbidity and zero mortality from veno-occlusive disease. Bone Marrow Transplantation. 2007,40(1),79-82.
[25]白海,欧英贤,达万明,等.自体骨髓移植与混合H LA半相合异体骨髓的自体骨髓移植治疗恶性血液病的比较.西北国防医学杂志.2000,1：21
[26]张奥莉,宋永平,魏旭东,等.复方丹参注射液预防肝静脉闭塞症58例报告.白血病淋巴瘤.2003,012(003)：146-147.
[27]Shulman HM, Fisher LB, Schoch HG, Henne KW, McDonald GB. Veno-occlusive disease of the liver after marrow transplantation:histological correlates of clinical signs and symptoms.Hepatology.1994,19:1171-1181.
[28]Deleve, l.D., shulman, H.m.,& mcDonald, g.B..Toxic injury to hepatic sinusoids:
sinusoidal obstruction syndrome (veno-occlusive disease). Seminars in Liver Disease.2002,22(1),27-42.
[29]DeLeve LD, Ito Y, Bethea NW, et al. Embolization by sinusoidal lining cells obstructs the microcirculation in rat sinusoidal obstruction syndrome. Am J Physiol Gastrointest Liver Physiol.2003,284:G1045-52.
[30]Coppell, J.A., Brown, s.A.,& Perry, D.J. veno-occlusive disease:cytokines, genetics, and haemostasis. Blood Reviews.2003,17(3),63-70.
[31]Gharib, m.i., Bulley, s.r., Doyle, J.J.,& Wynn, r.F. venous occlusive disease in children. Thrombosis Research.2006,118(1),27-38.
[32]Iguchi, A., Kobayashi, r., yoshida, m., Kobayashi, K., matsuo, K., Kitajima, i., et al. vascular endothelial growth factor (VEGF) is one of the cytokines causative and predictive of hepatic veno-occlusive disease (VOD) in stem cell transplantation. Bone Marrow Transplantation.2001,27(11),1173-1180.
[33]Ohashi, K., J. Tanabe, et al. The Japanese multicenter open randomized trial of ursodeoxycholic acid prophylaxis for hepatic veno-occlusive disease after stem cell transplantation." Am J Hematol.2000,64(1):32-38.
[34]Ruutu, T., B. Eriksson, et al. Ursodeoxycholic acid for the prevention of hepatic complications in allogeneic stem cell transplantation." Blood.2002,100(6): 1977-1983.
[35]Tay, J., A. Tinmouth, et al. Systematic review of controlled clinical trials on the use of ursodeoxycholic acid for the prevention of hepatic veno-occlusive disease in hematopoietic stem cell transplantation. Biol Blood Marrow Transplant.2007, 13(2):206-217.
[36]Bras G, Jeliffe DB, Stuart KL. Veno-occlusive disease of the live with non-portal type of cirrhosis occuring in Jamaica. Arch Pathol,1954,57:285-300.
[37]Chen, M.-Y., J.-T. Cai, et al. Hepatic veno-occlusive disease associated with the use of Gynura segetum." Eur J Intern Med.2007,18(8):609.
[38]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics. 2006,23(1),11-25.
[39]Deeg, H. J., H. M. Shulman, et al. Marrow graft rejection and veno-occlusive disease of the liver in patients with aplastic anemia conditioned with cyclophosphamide and cyclosporine." Transplantation.1986,42(5):497-501.
[40]Copelan, E. A., S. L. Penza, et al. The influence of early transplantation, age, GVHD prevention regimen, and other factors on outcome of allogeneic transplantation for CML following BuCy." Bone Marrow Transplant.2000,26(10): 1037-1043.
[41]Hill, K. R. Hepatic veno-occlusive disease produced experimentally in rats by the injection of monocrotaline." Lancet.1958,1(7021):623.
[42]Perazzo J, Eizayaga F, Romay S, et al.An experimental model of liver damageand Portal hypertension induced by a single dose of monoerotaline.Hepatogastroent-erelogy 1999,46:432-435
[43]McDonald GB, Sharma P, Matthews DE, et al. Veno-occlusive disease of the liver after bone marrow transplantation:diagnosis, incidence and predis-posing factors. Hepatology 1984; 4:116-22.
[44]McDonald GB, Hinds MS, Fisher LD,et al. Veno-occlusive disease of the liver and multiorgan failure after bone marrow transplantation a cohort study of 355 patients. Ann Intern Med 1993;18:255-67.
[45]Jones RJ, Lee KSK, Beschomer WE,et al. Veno-occlusive disease of the liver following bone marrow transplantation. Transplantation 1987; 44:778-779
[46]Gooley, T.A., rajvanshi, P., schoch, H.g.,& mcDonald, g.B. serum bilirubin levels and mortality after myeloablative allogeneic hematopoietic cell transplantation. Hepatology.2005,41(2),345-352.
[47]Litzow, m.r., repoussis, P.D., schroeder, g., schembri-Wismayer, D., Batts, K.P., Anderson, P.m., et al. veno-occlusive disease of the liver after blood and marrow transplantation:Analysis of pre-and post transplant risk factors associated with severity and results of therapy with tissue plasminogen activator. Leukemia and Lymphoma.2002,43(11),2099-2107.
[48]Klinger KW,Winqvist R,Riccio A. Identification of regulatory sequences in type
I gene is located at region q21.3-q22 of chromosome 7 and genetically linked with cystic fibrosis.Proc Natl Acad Sci USA.1987,84:8548-52.
[49]Kooistra T.ven den ber j,tons A. Butyrate stimulates tissue-type plasminogen-activator synthesis in culture human endothelial cells. Biochem J,1987,247(3):605-12.
[50]Kreistensen PL,Larsson LI,Nielsen LS.Human endothelial cells contain one type of plasminogen activiator. FEBS Lett.l984,168:33-37.
[51]Sprengers ED,Akkerman JW,Jansen BGBlood platelet plasminogen activator inhibitor:two different pools of endothelial cells type plasminogen activator inhibitor in human blood.Thromb Haemost.l986,55:325-329.
[52]Iguchi, A., Kobayashi, r., yoshida, m., Kobayashi, K., matsuo, K., Kitajima, i., et al. vascular endothelial growth factor (VEGF) is one of the cytokines causative and predictive of hepatic veno-occlusive disease (vOD) in stem cell transplantation. Bone Marrow Transplantation.2001,27(11),1173-1180
[53]Verena Pihusch, Markus Pihusch, Marius Penovici, Hans J. Kolb,Erhard Hiller, Rudolf Pihusch. Transforming growth factor beta-1 released from platelets contributes to hypercoagulability in veno-occlusive disease following hematopoetic stem cell transplantation.Tthrombosis research.2005,116:233-240.
[1]Bras G, Jeliffe DB, Stuart KL. Veno-occlusive disease of the live with non-portal type of cirrhosis occuring in Jamaica. Arch Pathol,1954,57:285-300.
[2]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics. 2006,23(1),11-25.
[3]Kumar S, DeLeve LD, Kamath PS, Tefferi A. Hepatic veno-occlusive disease (sinusoidal obstruction syndrome) after hematopoietic stem cell transplantation. Mayo Clin Proc 2003; 78:589-598
[4]McDonald GB, Hinds MS, Fisher LD et al. Veno-occlusive disease of the liver and multiorgan failure after bone marrow transplantation:a cohort study of 355 patients. Ann Intern Med 1993; 118:255-267.
[5]DeLeve LD. Cellular target of cyclophosphamide toxicity in the murine liver:role of glutathione and site of metabolic activation.Hepatology 1996; 24:830-837
[6]Meresse V, Hartmann O, Vassal G et al. Risk factors for hepatic veno-occlusive disease after high-dose busulfan-containing regimens followed by autologous bone marrow transplantation:a study in 136 children. Bone Marrow Transplant 1992;10:135-141
[7]Carreras E, Bertz H, Arcese W, et al, European Group for Blood and Marrow Transplantation Chronic Leukemia Working Party. Incidence and outcome of hepatic veno-occlusive disease after blood or marrow transplantation:a prospective cohort study of the European Group for Blood and Marrow Transplantation. Blood.1998;92:3599-3604.
[8]Lzaki T, Inomata Y, Asonuma K, et al. Early graft failure due to a veno-occlusive disease after a pediatric living donor liver transplantation[J]. Pediatr Transplant,2004,8(3):301.
[9]Jones RJ, Lee KS, BeschornerWE et al. Venoocclusive disease of the liver following bone marrow transplantation. Transplantation 1987; 44:778-783.
[10]DeLeve LD, McCuskey RS, Wang X, et al. Characterization of a reproducible rat model of hepatic veno-occlusive disease. Hepatology.1999;29:1779-1791
[11]高新生,肖绍树,贺降福.土三七生物碱成分分析及肝小静脉闭塞病小鼠模型建立.中国中西医结合杂志[J],2006,14(5)：311-313.
[12]Kallianpur AR, Hall LD, Yadav M, et al. The hemochromatosis C282Y allele:a risk factor for hepatic veno-occlusive disease after hematopoietic stem cell transplantation. Bone Marrow Transplant 2005; 35:1155-64.
[13]Cavet J, Middleton PG, Segall M, Noreen H, Davies SM, Dickinson AM. Recipient tumor necrosis factor-a and interleukin-10 gene polymorphisms associate with early mortality and acute graft-versus-host disease severity in HLA-matched sibling bone marrow transplants. Blood 1999; 94:3941-3946.
[14]Haire WD, Cavet J, Pavletic SZ. Tumor necrosis factor d3 allele predicts for organ dysfunction after allogeneic stem cell transplantation. Blood.2000; 96: 584a.
[15]Beutler E, Gelbart T, Kondo T, Matsunaga AT. The molecular basis of a case of c-glutamylcysteine synthetase deficiency. Blood.1999; 94:2890-2894.
[16]Poonkuzhali B, Vidya S, Shaji RV, Chandy M, Srivastava A.Glutathione S-transferase gene polymorphism and risk of hepatic venoocclusive disease in patients with thalassaemia major undergoing allogeneic bone marrow transplantation. Blood.2001; 98:852a.
[17]Shulman HM, Fisher LB, Schoch HG, Henne KW, McDonald GB. Veno-occlusive disease of the liver after marrow transplantation:histological correlates of clinical signs and symptoms.Hepatology.1994;19:1171-1181.
[18]DeLeve LD, Ito Y, Bethea NW, et al. Embolization by sinusoidal lining cells obstructs the microcirculation in rat sinusoidal obstruction syndrome. Am J Physiol Gastrointest Liver Physiol 2003; 284:G1045-52.
[19]Coppell, J.A., Brown, s.A.,& Perry, D.J. veno-occlusive disease:cytokines, genetics, and haemostasis. Blood Reviews.2003,17(3),63-70.
[20]Gharib, m.i., Bulley, s.r., Doyle, J.J.,& Wynn, r.F. venous occlusive disease in children. Thrombosis Research.2006,118(1),27-38.
[21]Iguchi, A., Kobayashi, r., yoshida, m., Kobayashi, K., matsuo, K., Kitajima, i., et
al. vascular endothelial growth factor (vEgF) is one of the cytokines causative and predictive of hepatic veno-occlusive disease (vOD) in stem cell transplantation. Bone Marrow Transplantation.2001,27(11),1173-1180.
[22]Shulman HM, Gown AM, Nugent DJ.Hepatic veno-occlusive disease after bone marrow transplantation-immunohistochemical identification of the material within occluded central venules. Am J Pathol.1987; 127:549-58.
[23]mcDonald, g.B., Hinds, m.s., Fisher, l.D., schoch, H.g., Wolford, J.l., Banaji, m., et al. veno-occlusive disease of the liver and multiorgan failure after bone marrow transplantation:A cohort study of 355 patients. Annals of Internal Medicine.. 1993,118(4),255-267.
[24]Bearman, s.i. The syndrome of hepatic veno-occlusive disease after marrow transplantation. Blood.1995,85(11),3005-3020.
[25]Gooley, T.A., rajvanshi, P., schoch, H.g.,& mcDonald, g.B. serum bilirubin levels and mortality after myeloablative allogeneic hematopoietic cell transplantation. Hepatology.2005,41(2),345-352.
[26]Litzow, m.r., repoussis, P.D., schroeder, g., schembri-Wismayer, D., Batts, K.P., Anderson, P.m., et al. veno-occlusive disease of the liver after blood and marrow transplantation:Analysis of pre-and post transplant risk factors associated with severity and results of therapy with tissue plasminogen activator. Leukemia and Lymphoma.2002,43(11),2099-2107.
[27]Bearman, s.i. The syndrome of hepatic veno-occlusive disease after marrow transplantation. Blood.1995,85(11),3005-3020.
[28]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics. 2006,23(1),11-25.
[29]Tanikawa S, Mori S, Ohhashi K, Akiyama H, Sasaki T, Kaku H, Hiruma K, Matsunaga T, Morita T, Sakamaki H. Predictive markers for hepatic veno-occlusive disease after hematopoietic stem cell transplantation in adults:a prospective single center study. Bone Marrow Transplant 2000; 26:881-886
[30]Deleve, I.D., shulman, H.m.,& mcDonald, g.B. Toxic injury to hepatic sinusoids: sinusoidal obstruction syndrome (veno-occlusive disease). Seminars in Liver Disease.2002,22(1),27-42.
[31]Eltumi M, Trivedi P, Hobbs JR, Portmann B, Cheeseman P, Downie C, Risteli J, Risteli L, Mowat AP. Monitoring of veno-occlusive disease after bone marrow transplantation by serum aminopropeptide of type Ⅲ procollagen. Lancet 1993; 342:518-521
[32]Rio, B., F. Bauduer, et al. N-Terminal Peptide of Type-Ⅰⅱ Procollagen-a Marker for the Development of Hepatic Venoocclusive Disease after Bmt and a Basis for Determining the Timing of Prophylactic Heparin. Bone Marrow Transplantation. 1993,11(6):471-472.
[33]Nurnberger, W., michelmann, i., Burdach, s.,& gobel, u. Endothelial dysfunction after bone marrow transplantation:increase of soluble thrombomodulin and PAi-1 in patients with multiple transplant-related complications. Annals of Hematology, 1998,76(2),61-65.
[34]Petaja J, Pitkanen S, Vettenranta K, Fasth A, Heikinheimo M. Serum tumor marker CA 125 is an early and sensitive indicator of veno-occlusive disease in children undergoing bone marrow transplantation. Clin Cancer Res 2000; 6: 531-535
[35]Carreras E, Garcia-Pagan JC, Bosch J, Rozman C. Transvenous liver biopsies in marrow transplant recipients. Transplantation 1995; 60:1375
[36]Shulman HM, Gooley T, Dudley MD, Kofl er T, Feldman R, Dwyer D, McDonald GB. Utility of transvenous liver biopsies and wedged hepatic venous pressure measurements in sixty marrow transplant recipients. Transplantation 1995; 59:1015-1022
[37]Senzolo M, Burra P, Cholongitas E, Lodato F, Marelli L, Manousou P, Patch D, Sturniolo GC, Burroughs AK. The transjugular route:the key hole to the liver world. Dig Liver Dis 2007; 39:105-116
[38]Bearman SI, Anderson GL, Mori M, Hinds MS, Shulman HM, McDonald GB. Venoocclusive disease of the liver:development of a model for predicting fatal outcome after marrow transplantation. J Clin Oncol 1993; 11:1729-1736
[39]Cholongitas E, Quaglia A, Samonakis D, Senzolo M, Triantos C, Patch D, Leandro G, Dhillon AP, Burroughs AK. Transjugular liver biopsy:how good is it for accurate histological interpretation? Gut 2006; 55:1789-1794
[40]Blostein MD, Paltiel OB, Thibault A, Rybka WB. A comparison of clinical criteria for the diagnosis of veno-occlusive disease of the liver after bone marrow transplantation. Bone Marrow Transplant 1992; 10:439-443
[41]McDonald GB, Sharma P, Matthews DE, et al. Veno-occlusive disease of the liver after bone marrow transplantation:diagnosis, incidence and predis-posing factors. Hepatology 1984; 4:116-22.
[42]McDonald GB, Hinds MS, Fisher LD,et al. Veno-occlusive disease of the liver and multiorgan failure after bone marrow transplantation a cohort study of 355 patients. Ann Intern Med 1993;18:255-67.
[43]Jones RJ, Lee KSK, Beschomer WE,et al. Veno-occlusive disease of the liver following bone marrow transplantation. Transplantation 1987; 44:778-779
[44]Corbacioglu, s., greil, J., Peters, c., Wulffraat, n., laws, H.J., Dilloo, D., et al. Deibrotide in the treatment of children with veno-occlusive disease (VOD):A retrospective multicentre study demonstrates therapeutic eficacy upon early intervention. Bone Marrow Transplant,2004,33(2),189-195.
[45]Bairey, O., Kirgner, i., yakobi, m., Hamdan, A., Ben-Ari, Z.,& shaklai, m. clinical severe hepatic veno-occlusive disease during induction treatment of acute monoblastic leukemia managed with deibrotide. American Journal of Hematology, 2002,69(4),281-284.
[46]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics, 2006,23(1),11-25.
[47]Richardson, P.g, Elias, A.D, Krishnan, A., Wheeler, c., nath, r., Hoppensteadt, D., et al. Treatment of severe veno-occlusive disease with deibrotide:compassionate use results in response without signiicant toxicity in a high-risk population. Blood, 1998,92(3),737-744.
[48]Dignan, F., gujral, D., Ethell, m., Evans, s., Treleaven, J., morgan, g., et al. Prophylactic deibrotide in allogeneic stem cell transplantation:minimal morbidity and zero mortality from veno-occlusive disease. Bone Marrow Transplantation, 2007,40(1),79-82.
[49]Benimetskaya, L., S. Wu, et al. Angiogenesis alteration by defibrotide: implications for its mechanism of action in severe hepatic veno-occlusive disease. 2008,Blood 112(10):4343-4352.
[50]Ho, V. T., C. Revta, et al. Hepatic veno-occlusive disease after hematopoietic stem cell transplantation:update on defibrotide and other current investigational therapies." Bone Marrow Transplant,2008,41(3):229-237.
[51]Qureshi, A., L. Marshall, et al. Defibrotide in the prevention and treatment of veno-occlusive disease in autologous and allogeneic stem cell transplantation in children." Pediatr Blood Cancer,2008,50(4):831-832.
[52]Imran, H., Tleyjeh, i.m., Zirakzadeh, A., rodriguez, v.,& Khan, s.P..use of prophylactic anticoagulation and the risk of hepatic veno-occlusive disease in patients undergoing hematopoietic stem cell transplantation:A systematic review and meta-analysis. Bone Marrow Transplantation,2006,37(7),677-686.
[53]Batsis, i., yannaki, E., Kaloyannidis, P., sakellari, i., smias, c., georgoulis, i., et al. veno-occlusive disease prophylaxis with fresh frozen plasma and heparin in bone marrow transplantation. Thrombosis Research,2006,118(5),611-618.
[54]Bearman, s.i., lee, J.l., Baron, A.E.,& mcDonald, g.B. Treatment of hepatic veno-occlusive disease with recombinant human tissue plasminogen activator and heparin in 42 marrow transplant patients. Blood,1997,89(5),1501-1506.
[55]litzow, m.r., repoussis, P.D., schroeder, g., schembri-Wismayer, D., Batts, K.P., Anderson, P.m., et al. veno-occlusive disease of the liver after blood and marrow transplantation:Analysis of pre-and post-transplant risk factors associated with severity and results of therapy with tissue plasminogen activator. Leukemia and Lymphoma,2002,43(11),2099-2107.
[56]Bajwa, r.P., cant, A.J., Abinun, m., Flood, T.J., Hodges, s., Hale, J.P., et al. recombinant tissue plasminogen activator for treatment of hepatic veno-occlusive disease following bone marrow transplantation in children:Effectiveness and a
scoring system for initiating treatment. Bone Marrow Transplantation,2003,31(7), 591-597.
[57]Ringden, O., remberger, m., lehmann, s., Hentschke, P., mattsson, J., Klaesson, s., et al. N-acetylcysteine for hepatic veno-occlusive disease after allogeneic stem cell transplantation. Bone Marrow Transplant,2000,25(9),993-996.
[58]Tefferi, A., Kumar, s., Wolf, r.c., lacy, m.Q., inwards, D.J., gloor, J.m., et al. charcoal hemoiltration for hepatic veno-occlusive disease after hematopoietic stem cell transplantation. Bone Marrow Transplantation,2001,28(10),997-999.
[59]Al Beihany, A., H. Al Omar, et al. Successful treatment of hepatic veno-occlusive disease after myeloablative allogeneic hematopoietic stem cell transplantation by early administration of a short course of methylprednisolone. Bone Marrow Transplant.2008,41(3):287-291.
[60]Akyuz, C., K. CaClar, et al. High-dose methylprednisolone treatment of hepatic veno-occlusive disease in a child with Wilms tumor." Pediatr Hematol Oncol 2003,20(4):345-349.
[61]Sayer, H. G, U. Will, et al. Hepatic veno-occlusive disease (VOD) with complete occlusion of liver venules after tandem autologous stem cell transplantation-successful treatment with high-dose methylprednisolone and defibrotide." J Cancer Res Clin Oncol,2002,128(3):148-152.
[62]Khoury, H., D. Adkins, et al. Does early treatment with high-dose methylprednisolone alter the course of hepatic regimen-related toxicity? Bone Marrow Transplant,2000,25(7):737-743.
[2]Brice, K., B. Valerie, et al. Risk-adjusted monitoring of veno-occlusive disease following Bayesian individualization of busulfan dosage for bone marrow transplantation in paediatrics. Pharmacoepidemiol Drug Saf.2008,17(2):135-143.
[3]Cacchione, A., A. LeMaitre, et al.. "Risk factors for hepatic veno-occlusive disease:a retrospective unicentric study in 116 children auto grafted after a high-dose BU-thiotepa regimen." Bone Marrow Transplant.2008,42(7):449-454.
[4]Carreras, E., L. Rosinol, et al. "Veno-occlusive disease of the liver after high-dose cytoreductive therapy with busulfan and melphalan for autologous blood stem cell transplantation in multiple myeloma patients." Biol Blood Marrow Transplant.2007,13(12):1448-1454.
[5]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and The rapeutics. 2006,23(1),11-25.
[6]Membreno, F. E., J. Ortiz, et al. Liver transplantation for sinusoidal obstructive syndrome (veno-occlusive disease):case report with review of the literature and the UNOS database. Clin Transplant.2008,22(4):397-404.
[7]de Fontbrune, F. S., H. Mal, et al. Veno-occlusive disease of the liver after lung transplantation. Am J Transplant.2007,7(9):2208-2211.
[8]Du, H., D.-L. Yang, et al. Hepatic veno-occlusive disease after hematopoietic stem cell transplantation in 23 patients. Zhonghua Gan Zang Bing Za Zhi.2007,15(12):949-950.
[9]Chen, M.-Y., J.-T. Cai, et al. Hepatic veno-occlusive disease associated with the use of Gynura segetum. Eur J Intern Med.2007,18(8):609.
[10]Wu, X.-j., K. Zhang, et al. Clinical analysis of four cases of hepatic
veno-occlusive disease caused by Gynura segetum (Lour.) Merr.Zhonghua Gan Zang Bing Za Zhi.2007,15(2):151-153.
[11]BorowskaH,AieiszewskaE,CzaudemaP,et,al.Daetinomycin-induced Veno-occlus-ive disease in rats. The hepatoprotective action of amifostine. Evaluation in a light and electron microscope.Folia Morph-ologica.2004,63:91-93
[12]Deleve LD et al. Characterization of Reproducible Rat Model of Hepatic Veno-occlusive Disease.Hepatology.1999,29:1779-1791.
[13]Chen MY, Cai JT et al. Reliable experimental model of hepatic veno-occlusive disease cause by monocrotaline [J].Hepatobiliary Pancreat Dis Int.2008,7(4):395-400.
[14]Gao XS, Xiao SH,et al. Analysis of alkaloids in sedum aizoon and establishement of hepatic veno-occlusive model in mice. Chin J Integr Trad West Med Dig.2006,14:311-313.
[15]Bajwa, r.P., cant, A.J., Abinun, m., Flood, T.J., Hodges, s., Hale, J.P., et al. recombinant tissue plasminogen activator for treatment of hepatic veno-occlusive disease following bone marrow transplantation in children:Effectiveness and a scoring system for initiating treatment. Bone Marrow Transplantation.2003,31(7), 591-597.
[16]litzow, m.r., repoussis, P.D., schroeder, g., schembri-Wismayer, D., Batts, K.P., Anderson, P.m, et al..veno-occlusive disease of the liver after blood and marrow transplantation:Analysis of pre-and post-transplant risk factors associated with severity and results of therapy with tissue plasminogen activator. Leukemia and Lymphoma.2002,43(11),2099-2107.
[17]Batsis, i., yannaki, E., Kaloyannidis, P., sakellari, i., smias, c., georgoulis, i., et al. veno-occlusive disease prophylaxis with fresh frozen plasma and heparin in bone marrow transplantation. Thrombosis Research.2006,118(5),611-618.
[18]Imran, H., Tleyjeh, i.m., Zirakzadeh, A., rodriguez, v.,& Khan, s.P.use of prophylactic anticoagulation and the risk of hepatic veno-occlusive disease in patients undergoing hematopoietic stem cell transplantation:A systematic review and meta-analysis. Bone Marrow Transplantation.2006,37(7),677-686.
[19]Sun, X., W.-G. Hao, et al. Efficacy of low-dose heparin and prostaglandin El in the prevention of hepatic veno-occlusive disease after allogenic hematopoietic stem cell transplantation in children with beta-thalassemia major. Zhongguo Dang Dai Er Ke Za Zhi.200,9(4):343-346.
[20]Al Beihany, A., H. Al Omar, et al. Successful treatment of hepatic veno-occlusive disease after myeloablative allogeneic hematopoietic stem cell transplantation by early administration of a short course of methylprednisolone. Bone Marrow Transplant.2008,41(3):287-291.
[21]Ringden, O., remberger, m., lehmann, s., Hentschke, P., mattsson, J., Klaesson, s., et al. N-acetylcysteine for hepatic veno-occlusive disease after allogeneic stem cell transplantation. Bone Marrow Transplant.2000,25(9),993-996.
[22]Bairey, O., Kirgner, i., yakobi, m., Hamdan, A., Ben-Ari, Z.,& shaklai, m. clinical severe hepatic veno-occlusive disease during induction treatment of acute monoblastic leukemia managed with deibrotide. American Journal of Hematology.2002,69(4),281-284.
[23]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics. 2006,23(1),11-25.
[24]Dignan, F., gujral, D., Ethell, m., Evans, s., Treleaven, J., morgan, g., et al. Prophylactic deibrotide in allogeneic stem cell transplantation:minimal morbidity and zero mortality from veno-occlusive disease. Bone Marrow Transplantation. 2007,40(1),79-82.
[25]白海,欧英贤,达万明,等.自体骨髓移植与混合H LA半相合异体骨髓的自体骨髓移植治疗恶性血液病的比较.西北国防医学杂志.2000,1：21
[26]张奥莉,宋永平,魏旭东,等.复方丹参注射液预防肝静脉闭塞症58例报告.白血病淋巴瘤.2003,012(003)：146-147.
[27]Shulman HM, Fisher LB, Schoch HG, Henne KW, McDonald GB. Veno-occlusive disease of the liver after marrow transplantation:histological correlates of clinical signs and symptoms.Hepatology.1994,19:1171-1181.
[28]Deleve, l.D., shulman, H.m.,& mcDonald, g.B..Toxic injury to hepatic sinusoids:
sinusoidal obstruction syndrome (veno-occlusive disease). Seminars in Liver Disease.2002,22(1),27-42.
[29]DeLeve LD, Ito Y, Bethea NW, et al. Embolization by sinusoidal lining cells obstructs the microcirculation in rat sinusoidal obstruction syndrome. Am J Physiol Gastrointest Liver Physiol.2003,284:G1045-52.
[30]Coppell, J.A., Brown, s.A.,& Perry, D.J. veno-occlusive disease:cytokines, genetics, and haemostasis. Blood Reviews.2003,17(3),63-70.
[31]Gharib, m.i., Bulley, s.r., Doyle, J.J.,& Wynn, r.F. venous occlusive disease in children. Thrombosis Research.2006,118(1),27-38.
[32]Iguchi, A., Kobayashi, r., yoshida, m., Kobayashi, K., matsuo, K., Kitajima, i., et al. vascular endothelial growth factor (VEGF) is one of the cytokines causative and predictive of hepatic veno-occlusive disease (VOD) in stem cell transplantation. Bone Marrow Transplantation.2001,27(11),1173-1180.
[33]Ohashi, K., J. Tanabe, et al. The Japanese multicenter open randomized trial of ursodeoxycholic acid prophylaxis for hepatic veno-occlusive disease after stem cell transplantation." Am J Hematol.2000,64(1):32-38.
[34]Ruutu, T., B. Eriksson, et al. Ursodeoxycholic acid for the prevention of hepatic complications in allogeneic stem cell transplantation." Blood.2002,100(6): 1977-1983.
[35]Tay, J., A. Tinmouth, et al. Systematic review of controlled clinical trials on the use of ursodeoxycholic acid for the prevention of hepatic veno-occlusive disease in hematopoietic stem cell transplantation. Biol Blood Marrow Transplant.2007, 13(2):206-217.
[36]Bras G, Jeliffe DB, Stuart KL. Veno-occlusive disease of the live with non-portal type of cirrhosis occuring in Jamaica. Arch Pathol,1954,57:285-300.
[37]Chen, M.-Y., J.-T. Cai, et al. Hepatic veno-occlusive disease associated with the use of Gynura segetum." Eur J Intern Med.2007,18(8):609.
[38]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics. 2006,23(1),11-25.
[39]Deeg, H. J., H. M. Shulman, et al. Marrow graft rejection and veno-occlusive disease of the liver in patients with aplastic anemia conditioned with cyclophosphamide and cyclosporine." Transplantation.1986,42(5):497-501.
[40]Copelan, E. A., S. L. Penza, et al. The influence of early transplantation, age, GVHD prevention regimen, and other factors on outcome of allogeneic transplantation for CML following BuCy." Bone Marrow Transplant.2000,26(10): 1037-1043.
[41]Hill, K. R. Hepatic veno-occlusive disease produced experimentally in rats by the injection of monocrotaline." Lancet.1958,1(7021):623.
[42]Perazzo J, Eizayaga F, Romay S, et al.An experimental model of liver damageand Portal hypertension induced by a single dose of monoerotaline.Hepatogastroent-erelogy 1999,46:432-435
[43]McDonald GB, Sharma P, Matthews DE, et al. Veno-occlusive disease of the liver after bone marrow transplantation:diagnosis, incidence and predis-posing factors. Hepatology 1984; 4:116-22.
[44]McDonald GB, Hinds MS, Fisher LD,et al. Veno-occlusive disease of the liver and multiorgan failure after bone marrow transplantation a cohort study of 355 patients. Ann Intern Med 1993;18:255-67.
[45]Jones RJ, Lee KSK, Beschomer WE,et al. Veno-occlusive disease of the liver following bone marrow transplantation. Transplantation 1987; 44:778-779
[46]Gooley, T.A., rajvanshi, P., schoch, H.g.,& mcDonald, g.B. serum bilirubin levels and mortality after myeloablative allogeneic hematopoietic cell transplantation. Hepatology.2005,41(2),345-352.
[47]Litzow, m.r., repoussis, P.D., schroeder, g., schembri-Wismayer, D., Batts, K.P., Anderson, P.m., et al. veno-occlusive disease of the liver after blood and marrow transplantation:Analysis of pre-and post transplant risk factors associated with severity and results of therapy with tissue plasminogen activator. Leukemia and Lymphoma.2002,43(11),2099-2107.
[48]Klinger KW,Winqvist R,Riccio A. Identification of regulatory sequences in type
I gene is located at region q21.3-q22 of chromosome 7 and genetically linked with cystic fibrosis.Proc Natl Acad Sci USA.1987,84:8548-52.
[49]Kooistra T.ven den ber j,tons A. Butyrate stimulates tissue-type plasminogen-activator synthesis in culture human endothelial cells. Biochem J,1987,247(3):605-12.
[50]Kreistensen PL,Larsson LI,Nielsen LS.Human endothelial cells contain one type of plasminogen activiator. FEBS Lett.l984,168:33-37.
[51]Sprengers ED,Akkerman JW,Jansen BGBlood platelet plasminogen activator inhibitor:two different pools of endothelial cells type plasminogen activator inhibitor in human blood.Thromb Haemost.l986,55:325-329.
[52]Iguchi, A., Kobayashi, r., yoshida, m., Kobayashi, K., matsuo, K., Kitajima, i., et al. vascular endothelial growth factor (VEGF) is one of the cytokines causative and predictive of hepatic veno-occlusive disease (vOD) in stem cell transplantation. Bone Marrow Transplantation.2001,27(11),1173-1180
[53]Verena Pihusch, Markus Pihusch, Marius Penovici, Hans J. Kolb,Erhard Hiller, Rudolf Pihusch. Transforming growth factor beta-1 released from platelets contributes to hypercoagulability in veno-occlusive disease following hematopoetic stem cell transplantation.Tthrombosis research.2005,116:233-240.
[1]Bras G, Jeliffe DB, Stuart KL. Veno-occlusive disease of the live with non-portal type of cirrhosis occuring in Jamaica. Arch Pathol,1954,57:285-300.
[2]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics. 2006,23(1),11-25.
[3]Kumar S, DeLeve LD, Kamath PS, Tefferi A. Hepatic veno-occlusive disease (sinusoidal obstruction syndrome) after hematopoietic stem cell transplantation. Mayo Clin Proc 2003; 78:589-598
[4]McDonald GB, Hinds MS, Fisher LD et al. Veno-occlusive disease of the liver and multiorgan failure after bone marrow transplantation:a cohort study of 355 patients. Ann Intern Med 1993; 118:255-267.
[5]DeLeve LD. Cellular target of cyclophosphamide toxicity in the murine liver:role of glutathione and site of metabolic activation.Hepatology 1996; 24:830-837
[6]Meresse V, Hartmann O, Vassal G et al. Risk factors for hepatic veno-occlusive disease after high-dose busulfan-containing regimens followed by autologous bone marrow transplantation:a study in 136 children. Bone Marrow Transplant 1992;10:135-141
[7]Carreras E, Bertz H, Arcese W, et al, European Group for Blood and Marrow Transplantation Chronic Leukemia Working Party. Incidence and outcome of hepatic veno-occlusive disease after blood or marrow transplantation:a prospective cohort study of the European Group for Blood and Marrow Transplantation. Blood.1998;92:3599-3604.
[8]Lzaki T, Inomata Y, Asonuma K, et al. Early graft failure due to a veno-occlusive disease after a pediatric living donor liver transplantation[J]. Pediatr Transplant,2004,8(3):301.
[9]Jones RJ, Lee KS, BeschornerWE et al. Venoocclusive disease of the liver following bone marrow transplantation. Transplantation 1987; 44:778-783.
[10]DeLeve LD, McCuskey RS, Wang X, et al. Characterization of a reproducible rat model of hepatic veno-occlusive disease. Hepatology.1999;29:1779-1791
[11]高新生,肖绍树,贺降福.土三七生物碱成分分析及肝小静脉闭塞病小鼠模型建立.中国中西医结合杂志[J],2006,14(5)：311-313.
[12]Kallianpur AR, Hall LD, Yadav M, et al. The hemochromatosis C282Y allele:a risk factor for hepatic veno-occlusive disease after hematopoietic stem cell transplantation. Bone Marrow Transplant 2005; 35:1155-64.
[13]Cavet J, Middleton PG, Segall M, Noreen H, Davies SM, Dickinson AM. Recipient tumor necrosis factor-a and interleukin-10 gene polymorphisms associate with early mortality and acute graft-versus-host disease severity in HLA-matched sibling bone marrow transplants. Blood 1999; 94:3941-3946.
[14]Haire WD, Cavet J, Pavletic SZ. Tumor necrosis factor d3 allele predicts for organ dysfunction after allogeneic stem cell transplantation. Blood.2000; 96: 584a.
[15]Beutler E, Gelbart T, Kondo T, Matsunaga AT. The molecular basis of a case of c-glutamylcysteine synthetase deficiency. Blood.1999; 94:2890-2894.
[16]Poonkuzhali B, Vidya S, Shaji RV, Chandy M, Srivastava A.Glutathione S-transferase gene polymorphism and risk of hepatic venoocclusive disease in patients with thalassaemia major undergoing allogeneic bone marrow transplantation. Blood.2001; 98:852a.
[17]Shulman HM, Fisher LB, Schoch HG, Henne KW, McDonald GB. Veno-occlusive disease of the liver after marrow transplantation:histological correlates of clinical signs and symptoms.Hepatology.1994;19:1171-1181.
[18]DeLeve LD, Ito Y, Bethea NW, et al. Embolization by sinusoidal lining cells obstructs the microcirculation in rat sinusoidal obstruction syndrome. Am J Physiol Gastrointest Liver Physiol 2003; 284:G1045-52.
[19]Coppell, J.A., Brown, s.A.,& Perry, D.J. veno-occlusive disease:cytokines, genetics, and haemostasis. Blood Reviews.2003,17(3),63-70.
[20]Gharib, m.i., Bulley, s.r., Doyle, J.J.,& Wynn, r.F. venous occlusive disease in children. Thrombosis Research.2006,118(1),27-38.
[21]Iguchi, A., Kobayashi, r., yoshida, m., Kobayashi, K., matsuo, K., Kitajima, i., et
al. vascular endothelial growth factor (vEgF) is one of the cytokines causative and predictive of hepatic veno-occlusive disease (vOD) in stem cell transplantation. Bone Marrow Transplantation.2001,27(11),1173-1180.
[22]Shulman HM, Gown AM, Nugent DJ.Hepatic veno-occlusive disease after bone marrow transplantation-immunohistochemical identification of the material within occluded central venules. Am J Pathol.1987; 127:549-58.
[23]mcDonald, g.B., Hinds, m.s., Fisher, l.D., schoch, H.g., Wolford, J.l., Banaji, m., et al. veno-occlusive disease of the liver and multiorgan failure after bone marrow transplantation:A cohort study of 355 patients. Annals of Internal Medicine.. 1993,118(4),255-267.
[24]Bearman, s.i. The syndrome of hepatic veno-occlusive disease after marrow transplantation. Blood.1995,85(11),3005-3020.
[25]Gooley, T.A., rajvanshi, P., schoch, H.g.,& mcDonald, g.B. serum bilirubin levels and mortality after myeloablative allogeneic hematopoietic cell transplantation. Hepatology.2005,41(2),345-352.
[26]Litzow, m.r., repoussis, P.D., schroeder, g., schembri-Wismayer, D., Batts, K.P., Anderson, P.m., et al. veno-occlusive disease of the liver after blood and marrow transplantation:Analysis of pre-and post transplant risk factors associated with severity and results of therapy with tissue plasminogen activator. Leukemia and Lymphoma.2002,43(11),2099-2107.
[27]Bearman, s.i. The syndrome of hepatic veno-occlusive disease after marrow transplantation. Blood.1995,85(11),3005-3020.
[28]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics. 2006,23(1),11-25.
[29]Tanikawa S, Mori S, Ohhashi K, Akiyama H, Sasaki T, Kaku H, Hiruma K, Matsunaga T, Morita T, Sakamaki H. Predictive markers for hepatic veno-occlusive disease after hematopoietic stem cell transplantation in adults:a prospective single center study. Bone Marrow Transplant 2000; 26:881-886
[30]Deleve, I.D., shulman, H.m.,& mcDonald, g.B. Toxic injury to hepatic sinusoids: sinusoidal obstruction syndrome (veno-occlusive disease). Seminars in Liver Disease.2002,22(1),27-42.
[31]Eltumi M, Trivedi P, Hobbs JR, Portmann B, Cheeseman P, Downie C, Risteli J, Risteli L, Mowat AP. Monitoring of veno-occlusive disease after bone marrow transplantation by serum aminopropeptide of type Ⅲ procollagen. Lancet 1993; 342:518-521
[32]Rio, B., F. Bauduer, et al. N-Terminal Peptide of Type-Ⅰⅱ Procollagen-a Marker for the Development of Hepatic Venoocclusive Disease after Bmt and a Basis for Determining the Timing of Prophylactic Heparin. Bone Marrow Transplantation. 1993,11(6):471-472.
[33]Nurnberger, W., michelmann, i., Burdach, s.,& gobel, u. Endothelial dysfunction after bone marrow transplantation:increase of soluble thrombomodulin and PAi-1 in patients with multiple transplant-related complications. Annals of Hematology, 1998,76(2),61-65.
[34]Petaja J, Pitkanen S, Vettenranta K, Fasth A, Heikinheimo M. Serum tumor marker CA 125 is an early and sensitive indicator of veno-occlusive disease in children undergoing bone marrow transplantation. Clin Cancer Res 2000; 6: 531-535
[35]Carreras E, Garcia-Pagan JC, Bosch J, Rozman C. Transvenous liver biopsies in marrow transplant recipients. Transplantation 1995; 60:1375
[36]Shulman HM, Gooley T, Dudley MD, Kofl er T, Feldman R, Dwyer D, McDonald GB. Utility of transvenous liver biopsies and wedged hepatic venous pressure measurements in sixty marrow transplant recipients. Transplantation 1995; 59:1015-1022
[37]Senzolo M, Burra P, Cholongitas E, Lodato F, Marelli L, Manousou P, Patch D, Sturniolo GC, Burroughs AK. The transjugular route:the key hole to the liver world. Dig Liver Dis 2007; 39:105-116
[38]Bearman SI, Anderson GL, Mori M, Hinds MS, Shulman HM, McDonald GB. Venoocclusive disease of the liver:development of a model for predicting fatal outcome after marrow transplantation. J Clin Oncol 1993; 11:1729-1736
[39]Cholongitas E, Quaglia A, Samonakis D, Senzolo M, Triantos C, Patch D, Leandro G, Dhillon AP, Burroughs AK. Transjugular liver biopsy:how good is it for accurate histological interpretation? Gut 2006; 55:1789-1794
[40]Blostein MD, Paltiel OB, Thibault A, Rybka WB. A comparison of clinical criteria for the diagnosis of veno-occlusive disease of the liver after bone marrow transplantation. Bone Marrow Transplant 1992; 10:439-443
[41]McDonald GB, Sharma P, Matthews DE, et al. Veno-occlusive disease of the liver after bone marrow transplantation:diagnosis, incidence and predis-posing factors. Hepatology 1984; 4:116-22.
[42]McDonald GB, Hinds MS, Fisher LD,et al. Veno-occlusive disease of the liver and multiorgan failure after bone marrow transplantation a cohort study of 355 patients. Ann Intern Med 1993;18:255-67.
[43]Jones RJ, Lee KSK, Beschomer WE,et al. Veno-occlusive disease of the liver following bone marrow transplantation. Transplantation 1987; 44:778-779
[44]Corbacioglu, s., greil, J., Peters, c., Wulffraat, n., laws, H.J., Dilloo, D., et al. Deibrotide in the treatment of children with veno-occlusive disease (VOD):A retrospective multicentre study demonstrates therapeutic eficacy upon early intervention. Bone Marrow Transplant,2004,33(2),189-195.
[45]Bairey, O., Kirgner, i., yakobi, m., Hamdan, A., Ben-Ari, Z.,& shaklai, m. clinical severe hepatic veno-occlusive disease during induction treatment of acute monoblastic leukemia managed with deibrotide. American Journal of Hematology, 2002,69(4),281-284.
[46]Helmy, A. review article:updates in the pathogenesis and therapy of hepatic sinusoidal obstruction syndrome. Alimentary Pharmacology and Therapeutics, 2006,23(1),11-25.
[47]Richardson, P.g, Elias, A.D, Krishnan, A., Wheeler, c., nath, r., Hoppensteadt, D., et al. Treatment of severe veno-occlusive disease with deibrotide:compassionate use results in response without signiicant toxicity in a high-risk population. Blood, 1998,92(3),737-744.
[48]Dignan, F., gujral, D., Ethell, m., Evans, s., Treleaven, J., morgan, g., et al. Prophylactic deibrotide in allogeneic stem cell transplantation:minimal morbidity and zero mortality from veno-occlusive disease. Bone Marrow Transplantation, 2007,40(1),79-82.
[49]Benimetskaya, L., S. Wu, et al. Angiogenesis alteration by defibrotide: implications for its mechanism of action in severe hepatic veno-occlusive disease. 2008,Blood 112(10):4343-4352.
[50]Ho, V. T., C. Revta, et al. Hepatic veno-occlusive disease after hematopoietic stem cell transplantation:update on defibrotide and other current investigational therapies." Bone Marrow Transplant,2008,41(3):229-237.
[51]Qureshi, A., L. Marshall, et al. Defibrotide in the prevention and treatment of veno-occlusive disease in autologous and allogeneic stem cell transplantation in children." Pediatr Blood Cancer,2008,50(4):831-832.
[52]Imran, H., Tleyjeh, i.m., Zirakzadeh, A., rodriguez, v.,& Khan, s.P..use of prophylactic anticoagulation and the risk of hepatic veno-occlusive disease in patients undergoing hematopoietic stem cell transplantation:A systematic review and meta-analysis. Bone Marrow Transplantation,2006,37(7),677-686.
[53]Batsis, i., yannaki, E., Kaloyannidis, P., sakellari, i., smias, c., georgoulis, i., et al. veno-occlusive disease prophylaxis with fresh frozen plasma and heparin in bone marrow transplantation. Thrombosis Research,2006,118(5),611-618.
[54]Bearman, s.i., lee, J.l., Baron, A.E.,& mcDonald, g.B. Treatment of hepatic veno-occlusive disease with recombinant human tissue plasminogen activator and heparin in 42 marrow transplant patients. Blood,1997,89(5),1501-1506.
[55]litzow, m.r., repoussis, P.D., schroeder, g., schembri-Wismayer, D., Batts, K.P., Anderson, P.m., et al. veno-occlusive disease of the liver after blood and marrow transplantation:Analysis of pre-and post-transplant risk factors associated with severity and results of therapy with tissue plasminogen activator. Leukemia and Lymphoma,2002,43(11),2099-2107.
[56]Bajwa, r.P., cant, A.J., Abinun, m., Flood, T.J., Hodges, s., Hale, J.P., et al. recombinant tissue plasminogen activator for treatment of hepatic veno-occlusive disease following bone marrow transplantation in children:Effectiveness and a
scoring system for initiating treatment. Bone Marrow Transplantation,2003,31(7), 591-597.
[57]Ringden, O., remberger, m., lehmann, s., Hentschke, P., mattsson, J., Klaesson, s., et al. N-acetylcysteine for hepatic veno-occlusive disease after allogeneic stem cell transplantation. Bone Marrow Transplant,2000,25(9),993-996.
[58]Tefferi, A., Kumar, s., Wolf, r.c., lacy, m.Q., inwards, D.J., gloor, J.m., et al. charcoal hemoiltration for hepatic veno-occlusive disease after hematopoietic stem cell transplantation. Bone Marrow Transplantation,2001,28(10),997-999.
[59]Al Beihany, A., H. Al Omar, et al. Successful treatment of hepatic veno-occlusive disease after myeloablative allogeneic hematopoietic stem cell transplantation by early administration of a short course of methylprednisolone. Bone Marrow Transplant.2008,41(3):287-291.
[60]Akyuz, C., K. CaClar, et al. High-dose methylprednisolone treatment of hepatic veno-occlusive disease in a child with Wilms tumor." Pediatr Hematol Oncol 2003,20(4):345-349.
[61]Sayer, H. G, U. Will, et al. Hepatic veno-occlusive disease (VOD) with complete occlusion of liver venules after tandem autologous stem cell transplantation-successful treatment with high-dose methylprednisolone and defibrotide." J Cancer Res Clin Oncol,2002,128(3):148-152.
[62]Khoury, H., D. Adkins, et al. Does early treatment with high-dose methylprednisolone alter the course of hepatic regimen-related toxicity? Bone Marrow Transplant,2000,25(7):737-743.