板蓝根多糖对自体肝移植大鼠一氧化氮和内皮素-1的影响
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摘要
目的和意义:
肝脏缺血再灌注损伤(hepatic ischemia reperfusion injury, HIRI)存在于任何术式肝移植手术过程中,是造成原发性肝无功能的重要原因之一。导致HIRI的原因很多,目前多数认为其损伤机理主要与微循环障碍、氧自由基的产生、能量物质耗竭、钙离子超载、线粒体功能受损等因素有关。其中微循环障碍起重要作用,并决定最终肝组织的损伤程度。一氧化氮(nitric oxide, NO)和内皮素-1(endothelin-1, ET-1)是两种效用相反的血管活性物质,二者浓度失衡是HIRI微循环障碍的一个重要原因。因此,本实验在建立大鼠自体原位肝移植模型的基础上,探讨板蓝根多糖(indigowoad root polysaccharide,IRPS)对移植肝的影响及NO、ET-1含量变化与其关系,从而为临床实践提供参考,减轻术后的并发症,提高术后的生存质量和生存率。
方法:
SD大鼠,随机分为3组,NC组:正常对照组(n=32);AT组:自体肝移植组(n=32),术前2周持续给予蒸馏水30mg·kg-1·d-1灌胃处理;IRPS组:板蓝根多糖+自体肝移植组(n=32),术前2周持续给予板蓝根多糖30mg·kg - 1·d– 1灌胃处理。三组大鼠分别于术后1h、6h、12h、24h各取8只处死,经门静脉采血作血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、一氧化氮(NO)、内皮素-1(ET-1)的检测;光镜下观察肝细胞显微结构变化,透射电镜下观察肝细胞及线粒体的超微结构改变。
结果:
(1)大鼠手术成功的标准:大鼠即刻苏醒、翻身,并能维持有效的呼吸、心跳。无肝期控制为(20±2.2)min。
(2)血清ALT、AST的情况:与NC组比较,AT组和IRPS组术后各时相点ALT及AST都有不同程度的升高,差异有统计学意义(P<0.05);但AT组与IRPS组相比较,AT组各时相点升高更加显著,差异有统计学意义(P<0.05)。
(3)血清NO、ET-1的情况:与NC相比,AT和IRPS两组术后各时相点血清NO含量都有不同程度的降低,ET-1含量都有不同程度的升高,差异有统计学意义(P <0.05);但AT组与IRPS组相比较,AT组各时相点NO降低及ET-1升高更加显著,差异有统计学意义(P <0.05)。
(4)肝细胞显微结构:AT组术后1h,6h,12h,肝细胞中重度水肿,肝窦明显淤血狭窄,胞浆疏松,炎症细胞轻度增多,小叶结构基本完整;与AT组相比,IRPS组肝细胞仅表现为轻度水肿。24h后,AT组肝组织灶状坏死,汇管区及坏死灶淋巴细胞浸润;IRPS组24h仅见肝细胞弥漫性中度水肿,未见明显坏死灶,肝小叶结构完整。
(5)肝细胞线粒体超微结构:AT组各时段肝细胞浓缩,细胞间隙变大,电子密度增高,细胞核皱缩变形,染色质粗糙,细胞器明显密集化,线粒体肿胀明显,膜模糊不清,部分膜破裂,线粒体嵴疏松溶解,有大量空泡形成;IRPS组肝细胞形态基本正常,线粒体仅表现为轻度肿胀,线粒体排列整齐。
结论:板蓝根多糖可以减轻肝移植过程中的HIRI,其可能与提高NO水平、降低ET-1水平,减轻微循环障碍有关。
OBJECTIVE:
Hepatic ischemia-reperfusion injury (hepatic ischemia reperfusion injury, HIRI) has existed in any surgical procedure of liver transplantation, which is the one of the important reason of primary nonfunction. There are many facts that leads to HIRI, it is generally accepted that the mechanism is related to microcirculatory disturbances, oxygen free radical generation, depletion of energy, calcium overload, dysfunction of mitochondria. In recent years, studies suggest that microcirculatory disturbances play an important role which can determine the final liver tissue injury. NO and ET-1 are two opposite utility of vasoactive substances, the concentration disequilibrium of which is the significance reason for microcirculation disturbances. Therefore, this experiment is based on the model of orthopotic liver autotransplantation of rats. To investigate the effect on the transplanted liver mediated by indigowoad root polysaccharide (IRPS) after the operation and the change of NO, ET-1 content, which may offer the information for clinical practice, lessen postoperative complications, and elevate the postoperative quality of life and the survival rate.
METHODS:
SD rats were assigned into three groups randomly, including NC group: normal control group (n=32); AT Group: autotransplantation group (n = 32), before the operation the rats were administer distilled water (30mg/kg) through intragastric administration once per day and continued 14 days; IRPS Group: indigowoad root polysaccharide group (n = 32), the rats in this group were received indigowoad root polysaccharide (30mg/kg) at the same method. Blood samples were collected at 1, 6, 12, 24 hours after operation of three groups for examination of the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO), and endothelin -1 (ET-1); the micro-structure of liver cell under light microscope and the ultrastructural changes of liver cell mitochondria were determined in three groups.
RESULTS:
(1) Rats surgical success criteria: Rats instantly awake, stand up, and can maintain an effective breathing, heartbeat. anhepatic phase was controlled in 20±2.2 minutes.
(2) The status of the serum ALT and AST: As compared with NC group, the ALT and AST was increased at whole process in AT and IRPS group (p<0.05), but compared with IRPS group, the data increased more obviously at whole process in AT group (p<0.05).
(3) The status of the serum NO and ET-1: As compared with NC group, the ET-1 was increased at whole process in AT and IRPS group (p<0.05), but compared with IRPS group, the data increased more obviously at whole process in AT group (p<0.05). the serum content of NO was decreased in AT and IRPS group, compared with NC group, but the content of NO was more decreased in AT group than that in IRPS group at whole process.
(4) The micro-structure of hepatocytes: In postoperative 1h, 6h and 12h the hepatocytes of AT group was severe edema, sinus hepaticus congestion and narrow significantly, endochylema raritas, and mild increase in inflammatory cells. Compared with the AT group, IRPS group of liver cells showed only mild edema. In postoperative 24h, the hepatocytes of AT group was focal necrosis, and periportal lymphocyte infiltration and necrosis; compared with AT group, the IRPS group only show moderate diffuse edema of liver cells, no significant necrosis, and hepatic lobule structural integrity.
(5) The ultrastructural changes of hepatocytes and mitochondria: the hepatocytes in the AT group had concentrated; increased electron density as the cells began to condense, condensed or split nuclei and nucleoli, swollen mitochondria, unclear mitochondrial membranes, some disrupted membranes and mitochondrial cristae which became solubilized. Nevertheless, the appearance of hepatocytes in the IRPS group was much better than that in the AT group. The structure of the mitochondria in the IRPS group was normal and was much better than that in the AT group.
CONCLUTION:
Indigowoad root polysaccharide on hepatic ischemia reperfusion injury has a protective effect after orthotopic liver autotransplantation, which may be related to increase NO levels and lower levels of ET-1 to reduce the microcirculation disturbance.
肝脏缺血再灌注损伤(hepatic ischemia reperfusion injury, HIRI)存在于任何术式肝移植手术过程中,是造成原发性肝无功能的重要原因之一。导致HIRI的原因很多,目前多数认为其损伤机理主要与微循环障碍、氧自由基的产生、能量物质耗竭、钙离子超载、线粒体功能受损等因素有关。其中微循环障碍起重要作用,并决定最终肝组织的损伤程度。一氧化氮(nitric oxide, NO)和内皮素-1(endothelin-1, ET-1)是两种效用相反的血管活性物质,二者浓度失衡是HIRI微循环障碍的一个重要原因。因此,本实验在建立大鼠自体原位肝移植模型的基础上,探讨板蓝根多糖(indigowoad root polysaccharide,IRPS)对移植肝的影响及NO、ET-1含量变化与其关系,从而为临床实践提供参考,减轻术后的并发症,提高术后的生存质量和生存率。
方法:
SD大鼠,随机分为3组,NC组:正常对照组(n=32);AT组:自体肝移植组(n=32),术前2周持续给予蒸馏水30mg·kg-1·d-1灌胃处理;IRPS组:板蓝根多糖+自体肝移植组(n=32),术前2周持续给予板蓝根多糖30mg·kg - 1·d– 1灌胃处理。三组大鼠分别于术后1h、6h、12h、24h各取8只处死,经门静脉采血作血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、一氧化氮(NO)、内皮素-1(ET-1)的检测;光镜下观察肝细胞显微结构变化,透射电镜下观察肝细胞及线粒体的超微结构改变。
结果:
(1)大鼠手术成功的标准:大鼠即刻苏醒、翻身,并能维持有效的呼吸、心跳。无肝期控制为(20±2.2)min。
(2)血清ALT、AST的情况:与NC组比较,AT组和IRPS组术后各时相点ALT及AST都有不同程度的升高,差异有统计学意义(P<0.05);但AT组与IRPS组相比较,AT组各时相点升高更加显著,差异有统计学意义(P<0.05)。
(3)血清NO、ET-1的情况:与NC相比,AT和IRPS两组术后各时相点血清NO含量都有不同程度的降低,ET-1含量都有不同程度的升高,差异有统计学意义(P <0.05);但AT组与IRPS组相比较,AT组各时相点NO降低及ET-1升高更加显著,差异有统计学意义(P <0.05)。
(4)肝细胞显微结构:AT组术后1h,6h,12h,肝细胞中重度水肿,肝窦明显淤血狭窄,胞浆疏松,炎症细胞轻度增多,小叶结构基本完整;与AT组相比,IRPS组肝细胞仅表现为轻度水肿。24h后,AT组肝组织灶状坏死,汇管区及坏死灶淋巴细胞浸润;IRPS组24h仅见肝细胞弥漫性中度水肿,未见明显坏死灶,肝小叶结构完整。
(5)肝细胞线粒体超微结构:AT组各时段肝细胞浓缩,细胞间隙变大,电子密度增高,细胞核皱缩变形,染色质粗糙,细胞器明显密集化,线粒体肿胀明显,膜模糊不清,部分膜破裂,线粒体嵴疏松溶解,有大量空泡形成;IRPS组肝细胞形态基本正常,线粒体仅表现为轻度肿胀,线粒体排列整齐。
结论:板蓝根多糖可以减轻肝移植过程中的HIRI,其可能与提高NO水平、降低ET-1水平,减轻微循环障碍有关。
OBJECTIVE:
Hepatic ischemia-reperfusion injury (hepatic ischemia reperfusion injury, HIRI) has existed in any surgical procedure of liver transplantation, which is the one of the important reason of primary nonfunction. There are many facts that leads to HIRI, it is generally accepted that the mechanism is related to microcirculatory disturbances, oxygen free radical generation, depletion of energy, calcium overload, dysfunction of mitochondria. In recent years, studies suggest that microcirculatory disturbances play an important role which can determine the final liver tissue injury. NO and ET-1 are two opposite utility of vasoactive substances, the concentration disequilibrium of which is the significance reason for microcirculation disturbances. Therefore, this experiment is based on the model of orthopotic liver autotransplantation of rats. To investigate the effect on the transplanted liver mediated by indigowoad root polysaccharide (IRPS) after the operation and the change of NO, ET-1 content, which may offer the information for clinical practice, lessen postoperative complications, and elevate the postoperative quality of life and the survival rate.
METHODS:
SD rats were assigned into three groups randomly, including NC group: normal control group (n=32); AT Group: autotransplantation group (n = 32), before the operation the rats were administer distilled water (30mg/kg) through intragastric administration once per day and continued 14 days; IRPS Group: indigowoad root polysaccharide group (n = 32), the rats in this group were received indigowoad root polysaccharide (30mg/kg) at the same method. Blood samples were collected at 1, 6, 12, 24 hours after operation of three groups for examination of the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO), and endothelin -1 (ET-1); the micro-structure of liver cell under light microscope and the ultrastructural changes of liver cell mitochondria were determined in three groups.
RESULTS:
(1) Rats surgical success criteria: Rats instantly awake, stand up, and can maintain an effective breathing, heartbeat. anhepatic phase was controlled in 20±2.2 minutes.
(2) The status of the serum ALT and AST: As compared with NC group, the ALT and AST was increased at whole process in AT and IRPS group (p<0.05), but compared with IRPS group, the data increased more obviously at whole process in AT group (p<0.05).
(3) The status of the serum NO and ET-1: As compared with NC group, the ET-1 was increased at whole process in AT and IRPS group (p<0.05), but compared with IRPS group, the data increased more obviously at whole process in AT group (p<0.05). the serum content of NO was decreased in AT and IRPS group, compared with NC group, but the content of NO was more decreased in AT group than that in IRPS group at whole process.
(4) The micro-structure of hepatocytes: In postoperative 1h, 6h and 12h the hepatocytes of AT group was severe edema, sinus hepaticus congestion and narrow significantly, endochylema raritas, and mild increase in inflammatory cells. Compared with the AT group, IRPS group of liver cells showed only mild edema. In postoperative 24h, the hepatocytes of AT group was focal necrosis, and periportal lymphocyte infiltration and necrosis; compared with AT group, the IRPS group only show moderate diffuse edema of liver cells, no significant necrosis, and hepatic lobule structural integrity.
(5) The ultrastructural changes of hepatocytes and mitochondria: the hepatocytes in the AT group had concentrated; increased electron density as the cells began to condense, condensed or split nuclei and nucleoli, swollen mitochondria, unclear mitochondrial membranes, some disrupted membranes and mitochondrial cristae which became solubilized. Nevertheless, the appearance of hepatocytes in the IRPS group was much better than that in the AT group. The structure of the mitochondria in the IRPS group was normal and was much better than that in the AT group.
CONCLUTION:
Indigowoad root polysaccharide on hepatic ischemia reperfusion injury has a protective effect after orthotopic liver autotransplantation, which may be related to increase NO levels and lower levels of ET-1 to reduce the microcirculation disturbance.
引文
[1]王生兰.肝缺血再灌注损伤的机制及药物保护作用[J].青海医学院学报, 2005,26(1):63-66.
[2]潘承恩.病毒性肝炎、肝癌与肝移植[J].西安交通大学学报(医学版), 2006,27 (2) :105-108.
[3]张水军,龚瑾,朱长举,等.左旋精氨酸对无心跳大鼠供肝缺血再灌注损伤防护作用研究[J].郑州大学学报(医学版) , 2004, 39(3) :407-409.
[4] Uhlmann D, Pietsch UC, Ludwig S, et al. Assessment of hepatic ischemia reperfusion injury by simultaneous measurement of tissue PO2 , PCO2 and pH [J]. Microvasc Res, 2004,67 (1) :38-47.
[5] Cutrn JC, Perrelli MG, Cavalieri B, et al. Microvascular dysfunction induced by reperfusion injury and protective effect of ischemic preconditioning [J]. Free Radical Biology & Medicine, 2002,33 (9):1200-1208.
[6] Serracino-Inglott F, Habib NA, Mathie RT. Hepatic ischemia-reperfusion injury [J]. Am J Surg, 2001,181 (2):160-166.
[7]李绍强,梁力建. L-精氨酸对肝缺血再灌注损伤的保护作用及其机制[J].中华实验外科杂志, 2003, 20: 320-321.
[8]孙德强.肝缺血-再灌注损害[J].国外医学外科学分册, 2002,29(4):206-207.
[9] Carden DL, Granger DN. Pathophysiology of ischemia-reperfusion injury [J]. J Pathol, 2000,190 (3):255-266.
[10] Garcia-Pagan JC, Zhang JX, Sonin N, et al. Ischemia / reperfusion induces an increase in the hepatic portal vasoconstrictive response to endothelin-1 [J]. Shock, 1999, 11: 325-329.
[11]陶立德,张培建,田明祥,等.低氧预适应对移植肝脏肝窦内皮细胞损伤的影响[J].实用临床医药杂志, 2007,11(2):39-42.
[12]张培建,卜平,金成.低氧预适应对移植大鼠肝细胞线粒体超微结构的影响[J].解剖学报, 2008,39(6):854-857.
[13]金成,张培建,李勇,等.低氧预适应对自体肝移植大鼠Bcl-2蛋白表达的影响及其意义[J].中国普通外科杂志,2009,11(1):31-35.
[14]张培建,周斌,金成,等.低氧预适应对移植肝内胆管的显微及超微结构的影响[J].中国现代普通外科进展,2008,11(6):470-473.
[15]周斌,张培建,李勇,等. VEGF参与低氧预适应对移植肝肝内胆管上皮细胞的保护作用[J].中国普外基础与临床杂志,2009,16(5):365-369.
[16]冯敏,刘新颜,张培建,等.去铁敏预处理对移植供肝保护作用的研究[J].中国普通外科杂志,2009,18(7):673-676.
[17]李吉萍,江鸿,刘墨祥.板蓝根多糖减肥调血脂与抗氧化作用研究[A].中国药理学会第九次全国会员代表大会暨全国药理学术会议论文集[C], 2007.
[18]张红英,王亚宾,王学兵,等.板蓝根多糖对体外培养的猪脾脏淋巴细胞增殖及分泌细胞因子和NO的影响[J].河南农业大学学报, 2009,43(2):85-83.
[19]张萍,刘捷,邓扬悟,等.板蓝根多糖的提取工艺及其清除自由基作用的初步研究[J].河南工业大学学报(自然科学版), 2006,27(3):36-38.
[20]朱道玉.板蓝根多糖对中华鳖肾脏抗氧化性的影响[J].西南师范大学学报(自然科学版), 2009,34(4):98-10.
[21]朱道玉.板蓝根多糖对中华鳖组织中超氧化物歧化酶活性的影响[J].动物医学进展, 2009,30(8):55-57.
[22]朱道玉.板蓝根多糖对中华鳖小肠抗氧化酶活性和脂质过氧化的影响[J].四川动物, 2009, 28(4):542-544.
[23]金成,张培建,冯敏,等.经门静脉灌注法大鼠自体原位肝移植模型的建立[J].中国普外基础与临床杂志, 2009,16(3):184-186.
[24]黄洁夫.肝脏移植的理论与实践[M].广州:广东科技出版社,1998,1.
[25] Aiba M, Takeyoshi I, Ohwade S, et a1. Novel nitric oxide donor (FK409) ameliorates liver damage during extended liver resection with warm ischemia in dogs [J]. J Am Coll Surg, 2001,193(3):264-271.
[26] Geller DA, Chia SH, Takahashi Y, et a1. Protective role of the L-arginine-nitric oxide synthase pathway on preservation injury after rat liver transplantation [J]. J Parenter Enteral Nutr, 2001,25(3): 142-147.
[27] Uhlmann D, Uhlmann S, Spiegel HU. Important role for endothelins in acute hepatic ischemia/reperfusion injury [J]. J Invest Surg, 2001,14(1):31-45.
[28] Ricciardi R, Schaffer BK, Shah SA, et a1. Bosentan, an endothelin antagonist, augments hepatic graft function by reducing graft circulatory impairment following ischemia/reperfusion injury [J]. J Gastrointest Surg, 2001,5(3):322-329.
[29] Marzi I, Takei Y, Ruker M, et al. Endothelin-1 is involvedin hepatic sinusoidal vasoconstriction after ischemia and reperfusion [J]. Transplant Int,1994,7(Suppl 1):S503-506.
[30] Koeppel TA, Thies JC, Schemmer P, et al. Inhibition of nitricoxide synthesis in ischemia/reperfusion of the rat liver isfollowed by impairment of hepatic microvascular blood flow [J]. J Hepatol, 1997,27(1):163-169.
[31] Takada Y, Bondjema K, Taeck D, et al. Platelet activatingfactor antagonist has a protective affect on preservation-reperfusioninjury of the graft in pig liver transplantation [J]. TransplantProc, 1995,27(1):747-748.
[32]齐欣,车向前,王超,等.中药预处理对肝脏缺血再灌注损伤保护作用的研究进展[J].中国普通外科杂志,2008,17(1):97-99.
[1] Uhlmann D, Pietsch UC , Ludwig S , et al. Assessment of hepatic ischemia reperfusion injury by simultaneous measurement of tissue PO2 , PCO2 and pH [J]. Microvasc Res, 2004,67 (1) :38-47.
[2] Cutrn JC, Perrelli MG , Cavalieri B, et al. Microvascular dysfunction induced byreperfusion injury and protective effect of ischemic preconditioning [J]. Free Radical Biology & Medicine, 2002,33 (9) :1200-1208.
[3] Ser racino-Inglott F, Habib NA, Mathie RT. Hepatic ischemia-reperfusion injury [J]. Am J Surg, 2001,181 (2):160-166.
[4]闵清,白育庭,吴基良,等.山楂叶总黄酮对心肌缺血再灌注损伤大鼠心功能的影响[J] .中国药学杂志,2005,40 (7):515-517.
[5] Yan Y, Davani S, Chocron S, et al. Effects of L-arginine administration before cardioplegic arrest on ischemia reprfusion injury [J]. Ann Thorac Surg, 2001,72 (6) :1985 -1990.
[6] Desrois M, Sciaky M, Lan C, et al. L-arginine during long-term ischemia: effects on cardiac fuction, energetic metabolism and endothelial damage [J]. J Heart Lung Transplant, 2000,19(4) :367 - 376.
[7] Yanagisawa M, Kurihura H, Kimura S et a1. 1988.A novel potent vasoconstrictor peptide produced by vascular endothelial cells [J]. Nature, 332(6163):411-415.
[8] Kawada N, Tran T, Klein H, et al. The contraction of hepatic stellate(Ito) cells stimulated with vasoactive substances:possible involvement of endothein 1 and nitric oxide in the regulation of the sinusoidal tonus [J]. Eur J Biochem, 1993,213(2):815-823.
[9] Billiar TR, Harbrecht BG.. Resolving the nitric oxide paradox in acute tissue damage [J]. Gastroenterology,1997,113(4):1405-1407.
[10] Clancy R, Rediske J, Koehne C, et al. Activation of stress-activated protein kinase in osteoarthritic cartilage: evidence for nitric oxide dependence [J]. Osteoarthritis Cartilage, 2001, 9(4):294-299.
[11] Kim Y, Lee S. Role of Kupffer cells in the vasoregulatory gene expression during hepatic ischemia/reperfusion [J]. Arch Pharm Res,2004,27(1):111-117.
[12] Carden DL, Granger DN. Pathophysiology of ischaemia-reperfusion injury [J]. J Pathol, 2000,190 (3):255-266.
[13] Garcia-Pagan JC, Zhang JX, Sonin N, et al. Ischemia / reperfusion induces an increase in the hepatic portal vasoconstrictive response to endothelin-1 [J]. Shock, 1999, 11(5): 325-329.
[14] Cingolani, Horacio E, Villa A, et al. The positive inotropic effect of angiotensin II: role of endothelin-1 and reactive oxygen species [J]. Hypertension, 2006,47(4):727-734.
[15] Galiuto, Leonarda MD, DeMaria, et al. Ischemia-reperfusion injury at the microvascular level: treatment by endothelin A– selective antagonist and evaluateon by myocardial contrast echocardiogramphy [J]. Circulation. 2000, 102(25):3111-3116.
[16] Peralta C, Bulbena O, Bargallo R, et al. Strategies to modulate the deleterious effects of endothelin in hepatic ischemia-reperfusion [J]. Transplantation. 2000, 70(12):1761-1770.
[17] Tsuchiya Y, Suzuki S, Inaba K, et al. Impact of endothelin-1 on microcirculatory disturbance after partial hepatectomy under ischemia/reperfusion in thioacetamide-induced cirrhotic rats [J]. J Surg Res, 2003,111(1):100-108.
[18] El-Wahsh M, Seifalian AM, Fuller BJ, et a. Induction of adhesion moleculeexpression in liver ischaemia-reperfusion injury is associated with impaired hepatic parenchymal microcirculation [J] . Br J Surg ,2004,91 (8):1034-1039.
[19] Uhlmann D, Uhlmann S, Spiegel HU. Important role for endothelins in acute hepatic ischemia/reperfusion injury [J]. J Invest Surg, 2001,14(1):31-45.
[20] Ricciardi R, Schaffer BK, Shah SA, et a1. Bosentan, an endothelin antagonist, augments hepatic graft function by reducing graft circulatory impairment following ischemia/reperfusion injury [J]. J Gastrointest Surg, 2001,5(3):322-329.
[21] Marzi I, Takei Y, Ruker M, et al. Endothelin-1 is involved in hepatic sinusoidal vasoconstriction after ischemia and reperfusion [J]. Transplant Int,1994,7(Suppl 1):S503-506.
[22] Koeppel TA, Thies JC, Schemmer P, et al. Inhibition of nitric oxide synthesis in ischemia/reperfusion of the rat liver is followed by impairment of hepatic microvascular blood flow [J]. J Hepatol, 1997,27(1):163-169.
[23] Takada Y, Bondjema K, Taeck D, et al. Platelet activating factor antagonist has a protective effect on preservation/reperfusion injury of the graft in pig liver transplantation [J]. Transplant Proc,1995,27(1):747-748.
[1] Kaiser AM, Saluja AK, Sengupta A, et al. Relationship between severity, necrosis and apoptosis in five models of experimental acute pancreatitis [J]. Physiology, 1995,269(5 Pt 1):1295-1304.
[2]吴宝强,秦锡虎,江勇,等.大鼠胰腺炎模型中腺泡细胞凋亡的观察[J].肝胆胰外科杂志, 2005,17(2):123-125.
[3] Kaiser AM, Saluja AK, Lu L, et al. Effects of cycloheximide on pancreatic endonuclease activity, apoptosis, and severity of acute pancreatitis [J]. Physiology, 1996,271(3 Pt 1):982-993.
[4]吴彪,王春友.细胞凋亡在实验性急性胰腺炎发病中的作用[J].中国普通外科杂志, 2004,13(10):732-735.
[5]周五一.凋亡相关基因Bax,Bcl-2在大鼠急性胰腺炎中的表达[J].空军总医院学报, 2002,18(4):223-226.
[6] Mareninova OA, Sung KF, Hong P, et al. Cell death in pancreatitis: caspases protect from necrotizing pancreatitis [J]. J Biol Chem, 2006 ,281(6):3370-3381.
[7] Frossard JL, Rubbia-Brandt L, Wallig MA, et al. Severe acute pancreatitis and reduced acinar cell apoptosis in the exocrine pancreas of mice deficient for the Cx32 gene [J]. Gastroenterology, 2003,124(2):481-493.
[8]范志宁,刘训良,熊观瀛,等.急性梗阻性胰腺炎大鼠胰腺腺泡细胞凋亡的变化[J].世界华人消化杂志, 2006,14(9):912-915.
[9]朱人敏,张晓华,李兆申,等. Caspase-1抑制剂对实验性重症急性胰腺炎的治疗作用[J].解放军医学杂志, 2005,30(4):283-285.
[10] Mews P, Philips P, Fahmy R, et al. Pancreatic stellate cells respond to inflammatory cytokines: potential role in chronic pancreatitis [J]. Gut, 2002,50(4):535-541.
[11] Elsing C, Harenberg S, Stremmel W, et al. Serum levels of soluble Fas, nitric oxide and cytokines in acute decompensated cirrhotic patients [J]. Gastroenterol, 2007,13(3):421-425.
[12]张永宏,周力,黄立敏.细胞凋亡和肿瘤坏死因子在大鼠急性胰腺炎发病中的作用[J].贵州医药, 2004,28(12):1081-1083.
[13] Malleo G, Mazzon E, Genovese T, et al. Etanercept attenuates the development of cerulein-induced acute pancreatitis in mice: a comparison with TNF-alpha genetic deletion [J]. Shock, 2007,27(5):542-551.
[14]黄旻,吴小翎.细胞因子与重症急性胰腺炎[J].中国煤炭工业医学杂志,2006,9(10):1026-1027.
[15]范昕,张建新,瞿建国,等.白细胞介素1受体拮抗剂对胰腺炎大鼠胰腺细胞凋亡的诱[J].江苏医药, 2007,33(6):619-620.
[16]毛正发,张建新,瞿建国,等.白细胞介素1受体拮抗剂诱导急性胰腺炎大鼠胰腺腺泡细胞凋亡机制的研究[J].江苏大学学报(医学版) , 2007,17(5):396-399.
[17]张国伟,周杰,廖彩仙,等.细胞核转录因子κB在慢性胰腺炎中表达的研究[J].中国普通外科杂志, 2005,14(11):860-862.
[18] Sathyanarayan G, Garg PK, Prasad H, et al. Elevated level of interleukin-6 predicts organ failure and severe disease in patients with acute pancreatitis [J]. Gastroenterol Hepatol, 2007,22(4):550-554.
[19] Gulcubuk A, Altunatmaz K, Aonmez K, et al. Effects of curcumin on tumor necrosis factor-alpha and interleukin-6 in the late phase of experimental acute pancreatitis [J]. Vet Med A Physiol Pathol Clin Med, 2006,53(1):49-54.
[20] Chao KC, Chao KF, Chuang CC, et al. Blockade of interleukin-6 accelerates acinar cell apoptosis and attenuates experimental acute pancreatitis [J]. Surg, 2006,93(3):332-338.
[21] Gu C, Wang Y, Zhang ZT, et al. Effects of human interleukin 10 gene transfer on the expression of Bcl-2 Bax and apoptosis of hepatocyte in rats with acute hemorrhangic necrotizing pancreatitis [J]. Chin Med, 2005,118(9):1658-1660.
[22] Cao Y, Adhikari S, Clément MV, et al. Induction of apoptosis by crambene protects mice against acute pancreatitis via anti-inflammatory pathways [J]. Pathol, 2007, 170(5):1521-1534.
[23] Takeyama Y, Hori Y ,Takase K ,et al . Apoptotic cell death of hepatocytes in rat experimental severe acute pancreatitis [J]. Surgery, 2000,127(1):55-64.
[24] Matsuzaki S, Ito M, Onizuka S, et al. Corelation between transforming growth factor beta expression and apoptosis in the rat pancreas during regenerayipn from caerulein induced pancreatitis [J] .Digestion, 2004,58(2):210-218.
[25] Nagashio Y, Ueno H, Imamura M, et al. Inhibition of transforming growth factor beta decreases pancreatic fibrosisand protects the pancreas against chronic injury in mice [J]. Lab Invest, 2004,84(12):1610-1618.
[2]潘承恩.病毒性肝炎、肝癌与肝移植[J].西安交通大学学报(医学版), 2006,27 (2) :105-108.
[3]张水军,龚瑾,朱长举,等.左旋精氨酸对无心跳大鼠供肝缺血再灌注损伤防护作用研究[J].郑州大学学报(医学版) , 2004, 39(3) :407-409.
[4] Uhlmann D, Pietsch UC, Ludwig S, et al. Assessment of hepatic ischemia reperfusion injury by simultaneous measurement of tissue PO2 , PCO2 and pH [J]. Microvasc Res, 2004,67 (1) :38-47.
[5] Cutrn JC, Perrelli MG, Cavalieri B, et al. Microvascular dysfunction induced by reperfusion injury and protective effect of ischemic preconditioning [J]. Free Radical Biology & Medicine, 2002,33 (9):1200-1208.
[6] Serracino-Inglott F, Habib NA, Mathie RT. Hepatic ischemia-reperfusion injury [J]. Am J Surg, 2001,181 (2):160-166.
[7]李绍强,梁力建. L-精氨酸对肝缺血再灌注损伤的保护作用及其机制[J].中华实验外科杂志, 2003, 20: 320-321.
[8]孙德强.肝缺血-再灌注损害[J].国外医学外科学分册, 2002,29(4):206-207.
[9] Carden DL, Granger DN. Pathophysiology of ischemia-reperfusion injury [J]. J Pathol, 2000,190 (3):255-266.
[10] Garcia-Pagan JC, Zhang JX, Sonin N, et al. Ischemia / reperfusion induces an increase in the hepatic portal vasoconstrictive response to endothelin-1 [J]. Shock, 1999, 11: 325-329.
[11]陶立德,张培建,田明祥,等.低氧预适应对移植肝脏肝窦内皮细胞损伤的影响[J].实用临床医药杂志, 2007,11(2):39-42.
[12]张培建,卜平,金成.低氧预适应对移植大鼠肝细胞线粒体超微结构的影响[J].解剖学报, 2008,39(6):854-857.
[13]金成,张培建,李勇,等.低氧预适应对自体肝移植大鼠Bcl-2蛋白表达的影响及其意义[J].中国普通外科杂志,2009,11(1):31-35.
[14]张培建,周斌,金成,等.低氧预适应对移植肝内胆管的显微及超微结构的影响[J].中国现代普通外科进展,2008,11(6):470-473.
[15]周斌,张培建,李勇,等. VEGF参与低氧预适应对移植肝肝内胆管上皮细胞的保护作用[J].中国普外基础与临床杂志,2009,16(5):365-369.
[16]冯敏,刘新颜,张培建,等.去铁敏预处理对移植供肝保护作用的研究[J].中国普通外科杂志,2009,18(7):673-676.
[17]李吉萍,江鸿,刘墨祥.板蓝根多糖减肥调血脂与抗氧化作用研究[A].中国药理学会第九次全国会员代表大会暨全国药理学术会议论文集[C], 2007.
[18]张红英,王亚宾,王学兵,等.板蓝根多糖对体外培养的猪脾脏淋巴细胞增殖及分泌细胞因子和NO的影响[J].河南农业大学学报, 2009,43(2):85-83.
[19]张萍,刘捷,邓扬悟,等.板蓝根多糖的提取工艺及其清除自由基作用的初步研究[J].河南工业大学学报(自然科学版), 2006,27(3):36-38.
[20]朱道玉.板蓝根多糖对中华鳖肾脏抗氧化性的影响[J].西南师范大学学报(自然科学版), 2009,34(4):98-10.
[21]朱道玉.板蓝根多糖对中华鳖组织中超氧化物歧化酶活性的影响[J].动物医学进展, 2009,30(8):55-57.
[22]朱道玉.板蓝根多糖对中华鳖小肠抗氧化酶活性和脂质过氧化的影响[J].四川动物, 2009, 28(4):542-544.
[23]金成,张培建,冯敏,等.经门静脉灌注法大鼠自体原位肝移植模型的建立[J].中国普外基础与临床杂志, 2009,16(3):184-186.
[24]黄洁夫.肝脏移植的理论与实践[M].广州:广东科技出版社,1998,1.
[25] Aiba M, Takeyoshi I, Ohwade S, et a1. Novel nitric oxide donor (FK409) ameliorates liver damage during extended liver resection with warm ischemia in dogs [J]. J Am Coll Surg, 2001,193(3):264-271.
[26] Geller DA, Chia SH, Takahashi Y, et a1. Protective role of the L-arginine-nitric oxide synthase pathway on preservation injury after rat liver transplantation [J]. J Parenter Enteral Nutr, 2001,25(3): 142-147.
[27] Uhlmann D, Uhlmann S, Spiegel HU. Important role for endothelins in acute hepatic ischemia/reperfusion injury [J]. J Invest Surg, 2001,14(1):31-45.
[28] Ricciardi R, Schaffer BK, Shah SA, et a1. Bosentan, an endothelin antagonist, augments hepatic graft function by reducing graft circulatory impairment following ischemia/reperfusion injury [J]. J Gastrointest Surg, 2001,5(3):322-329.
[29] Marzi I, Takei Y, Ruker M, et al. Endothelin-1 is involvedin hepatic sinusoidal vasoconstriction after ischemia and reperfusion [J]. Transplant Int,1994,7(Suppl 1):S503-506.
[30] Koeppel TA, Thies JC, Schemmer P, et al. Inhibition of nitricoxide synthesis in ischemia/reperfusion of the rat liver isfollowed by impairment of hepatic microvascular blood flow [J]. J Hepatol, 1997,27(1):163-169.
[31] Takada Y, Bondjema K, Taeck D, et al. Platelet activatingfactor antagonist has a protective affect on preservation-reperfusioninjury of the graft in pig liver transplantation [J]. TransplantProc, 1995,27(1):747-748.
[32]齐欣,车向前,王超,等.中药预处理对肝脏缺血再灌注损伤保护作用的研究进展[J].中国普通外科杂志,2008,17(1):97-99.
[1] Uhlmann D, Pietsch UC , Ludwig S , et al. Assessment of hepatic ischemia reperfusion injury by simultaneous measurement of tissue PO2 , PCO2 and pH [J]. Microvasc Res, 2004,67 (1) :38-47.
[2] Cutrn JC, Perrelli MG , Cavalieri B, et al. Microvascular dysfunction induced byreperfusion injury and protective effect of ischemic preconditioning [J]. Free Radical Biology & Medicine, 2002,33 (9) :1200-1208.
[3] Ser racino-Inglott F, Habib NA, Mathie RT. Hepatic ischemia-reperfusion injury [J]. Am J Surg, 2001,181 (2):160-166.
[4]闵清,白育庭,吴基良,等.山楂叶总黄酮对心肌缺血再灌注损伤大鼠心功能的影响[J] .中国药学杂志,2005,40 (7):515-517.
[5] Yan Y, Davani S, Chocron S, et al. Effects of L-arginine administration before cardioplegic arrest on ischemia reprfusion injury [J]. Ann Thorac Surg, 2001,72 (6) :1985 -1990.
[6] Desrois M, Sciaky M, Lan C, et al. L-arginine during long-term ischemia: effects on cardiac fuction, energetic metabolism and endothelial damage [J]. J Heart Lung Transplant, 2000,19(4) :367 - 376.
[7] Yanagisawa M, Kurihura H, Kimura S et a1. 1988.A novel potent vasoconstrictor peptide produced by vascular endothelial cells [J]. Nature, 332(6163):411-415.
[8] Kawada N, Tran T, Klein H, et al. The contraction of hepatic stellate(Ito) cells stimulated with vasoactive substances:possible involvement of endothein 1 and nitric oxide in the regulation of the sinusoidal tonus [J]. Eur J Biochem, 1993,213(2):815-823.
[9] Billiar TR, Harbrecht BG.. Resolving the nitric oxide paradox in acute tissue damage [J]. Gastroenterology,1997,113(4):1405-1407.
[10] Clancy R, Rediske J, Koehne C, et al. Activation of stress-activated protein kinase in osteoarthritic cartilage: evidence for nitric oxide dependence [J]. Osteoarthritis Cartilage, 2001, 9(4):294-299.
[11] Kim Y, Lee S. Role of Kupffer cells in the vasoregulatory gene expression during hepatic ischemia/reperfusion [J]. Arch Pharm Res,2004,27(1):111-117.
[12] Carden DL, Granger DN. Pathophysiology of ischaemia-reperfusion injury [J]. J Pathol, 2000,190 (3):255-266.
[13] Garcia-Pagan JC, Zhang JX, Sonin N, et al. Ischemia / reperfusion induces an increase in the hepatic portal vasoconstrictive response to endothelin-1 [J]. Shock, 1999, 11(5): 325-329.
[14] Cingolani, Horacio E, Villa A, et al. The positive inotropic effect of angiotensin II: role of endothelin-1 and reactive oxygen species [J]. Hypertension, 2006,47(4):727-734.
[15] Galiuto, Leonarda MD, DeMaria, et al. Ischemia-reperfusion injury at the microvascular level: treatment by endothelin A– selective antagonist and evaluateon by myocardial contrast echocardiogramphy [J]. Circulation. 2000, 102(25):3111-3116.
[16] Peralta C, Bulbena O, Bargallo R, et al. Strategies to modulate the deleterious effects of endothelin in hepatic ischemia-reperfusion [J]. Transplantation. 2000, 70(12):1761-1770.
[17] Tsuchiya Y, Suzuki S, Inaba K, et al. Impact of endothelin-1 on microcirculatory disturbance after partial hepatectomy under ischemia/reperfusion in thioacetamide-induced cirrhotic rats [J]. J Surg Res, 2003,111(1):100-108.
[18] El-Wahsh M, Seifalian AM, Fuller BJ, et a. Induction of adhesion moleculeexpression in liver ischaemia-reperfusion injury is associated with impaired hepatic parenchymal microcirculation [J] . Br J Surg ,2004,91 (8):1034-1039.
[19] Uhlmann D, Uhlmann S, Spiegel HU. Important role for endothelins in acute hepatic ischemia/reperfusion injury [J]. J Invest Surg, 2001,14(1):31-45.
[20] Ricciardi R, Schaffer BK, Shah SA, et a1. Bosentan, an endothelin antagonist, augments hepatic graft function by reducing graft circulatory impairment following ischemia/reperfusion injury [J]. J Gastrointest Surg, 2001,5(3):322-329.
[21] Marzi I, Takei Y, Ruker M, et al. Endothelin-1 is involved in hepatic sinusoidal vasoconstriction after ischemia and reperfusion [J]. Transplant Int,1994,7(Suppl 1):S503-506.
[22] Koeppel TA, Thies JC, Schemmer P, et al. Inhibition of nitric oxide synthesis in ischemia/reperfusion of the rat liver is followed by impairment of hepatic microvascular blood flow [J]. J Hepatol, 1997,27(1):163-169.
[23] Takada Y, Bondjema K, Taeck D, et al. Platelet activating factor antagonist has a protective effect on preservation/reperfusion injury of the graft in pig liver transplantation [J]. Transplant Proc,1995,27(1):747-748.
[1] Kaiser AM, Saluja AK, Sengupta A, et al. Relationship between severity, necrosis and apoptosis in five models of experimental acute pancreatitis [J]. Physiology, 1995,269(5 Pt 1):1295-1304.
[2]吴宝强,秦锡虎,江勇,等.大鼠胰腺炎模型中腺泡细胞凋亡的观察[J].肝胆胰外科杂志, 2005,17(2):123-125.
[3] Kaiser AM, Saluja AK, Lu L, et al. Effects of cycloheximide on pancreatic endonuclease activity, apoptosis, and severity of acute pancreatitis [J]. Physiology, 1996,271(3 Pt 1):982-993.
[4]吴彪,王春友.细胞凋亡在实验性急性胰腺炎发病中的作用[J].中国普通外科杂志, 2004,13(10):732-735.
[5]周五一.凋亡相关基因Bax,Bcl-2在大鼠急性胰腺炎中的表达[J].空军总医院学报, 2002,18(4):223-226.
[6] Mareninova OA, Sung KF, Hong P, et al. Cell death in pancreatitis: caspases protect from necrotizing pancreatitis [J]. J Biol Chem, 2006 ,281(6):3370-3381.
[7] Frossard JL, Rubbia-Brandt L, Wallig MA, et al. Severe acute pancreatitis and reduced acinar cell apoptosis in the exocrine pancreas of mice deficient for the Cx32 gene [J]. Gastroenterology, 2003,124(2):481-493.
[8]范志宁,刘训良,熊观瀛,等.急性梗阻性胰腺炎大鼠胰腺腺泡细胞凋亡的变化[J].世界华人消化杂志, 2006,14(9):912-915.
[9]朱人敏,张晓华,李兆申,等. Caspase-1抑制剂对实验性重症急性胰腺炎的治疗作用[J].解放军医学杂志, 2005,30(4):283-285.
[10] Mews P, Philips P, Fahmy R, et al. Pancreatic stellate cells respond to inflammatory cytokines: potential role in chronic pancreatitis [J]. Gut, 2002,50(4):535-541.
[11] Elsing C, Harenberg S, Stremmel W, et al. Serum levels of soluble Fas, nitric oxide and cytokines in acute decompensated cirrhotic patients [J]. Gastroenterol, 2007,13(3):421-425.
[12]张永宏,周力,黄立敏.细胞凋亡和肿瘤坏死因子在大鼠急性胰腺炎发病中的作用[J].贵州医药, 2004,28(12):1081-1083.
[13] Malleo G, Mazzon E, Genovese T, et al. Etanercept attenuates the development of cerulein-induced acute pancreatitis in mice: a comparison with TNF-alpha genetic deletion [J]. Shock, 2007,27(5):542-551.
[14]黄旻,吴小翎.细胞因子与重症急性胰腺炎[J].中国煤炭工业医学杂志,2006,9(10):1026-1027.
[15]范昕,张建新,瞿建国,等.白细胞介素1受体拮抗剂对胰腺炎大鼠胰腺细胞凋亡的诱[J].江苏医药, 2007,33(6):619-620.
[16]毛正发,张建新,瞿建国,等.白细胞介素1受体拮抗剂诱导急性胰腺炎大鼠胰腺腺泡细胞凋亡机制的研究[J].江苏大学学报(医学版) , 2007,17(5):396-399.
[17]张国伟,周杰,廖彩仙,等.细胞核转录因子κB在慢性胰腺炎中表达的研究[J].中国普通外科杂志, 2005,14(11):860-862.
[18] Sathyanarayan G, Garg PK, Prasad H, et al. Elevated level of interleukin-6 predicts organ failure and severe disease in patients with acute pancreatitis [J]. Gastroenterol Hepatol, 2007,22(4):550-554.
[19] Gulcubuk A, Altunatmaz K, Aonmez K, et al. Effects of curcumin on tumor necrosis factor-alpha and interleukin-6 in the late phase of experimental acute pancreatitis [J]. Vet Med A Physiol Pathol Clin Med, 2006,53(1):49-54.
[20] Chao KC, Chao KF, Chuang CC, et al. Blockade of interleukin-6 accelerates acinar cell apoptosis and attenuates experimental acute pancreatitis [J]. Surg, 2006,93(3):332-338.
[21] Gu C, Wang Y, Zhang ZT, et al. Effects of human interleukin 10 gene transfer on the expression of Bcl-2 Bax and apoptosis of hepatocyte in rats with acute hemorrhangic necrotizing pancreatitis [J]. Chin Med, 2005,118(9):1658-1660.
[22] Cao Y, Adhikari S, Clément MV, et al. Induction of apoptosis by crambene protects mice against acute pancreatitis via anti-inflammatory pathways [J]. Pathol, 2007, 170(5):1521-1534.
[23] Takeyama Y, Hori Y ,Takase K ,et al . Apoptotic cell death of hepatocytes in rat experimental severe acute pancreatitis [J]. Surgery, 2000,127(1):55-64.
[24] Matsuzaki S, Ito M, Onizuka S, et al. Corelation between transforming growth factor beta expression and apoptosis in the rat pancreas during regenerayipn from caerulein induced pancreatitis [J] .Digestion, 2004,58(2):210-218.
[25] Nagashio Y, Ueno H, Imamura M, et al. Inhibition of transforming growth factor beta decreases pancreatic fibrosisand protects the pancreas against chronic injury in mice [J]. Lab Invest, 2004,84(12):1610-1618.