DMN诱发大鼠肝纤维化库普弗细胞与肝星状细胞的分布
|
摘要
目的:探讨二甲基亚硝胺(DMN)诱发大鼠肝纤维化库普弗细胞与肝星状细胞的分布及其意义。方法:选取40只Wistar雄性大鼠连续4周腹腔注射10 g/L DMN制备肝纤维化模型。采用随机数字表法分为模型组(n=20)与正常组(n=20),两组均应用DMN进行肝纤维化造模。采用Reiman法测定大鼠第4周和第7周ALT、AST水平,免疫组化法测定两组大鼠库普弗细胞与肝星状细胞标志物ED1与α-SMA分布情况,收集并分析两组肝/体质量比、肝组织病理变化、胶原纤维面密度。结果:模型组第4周和第7周ALT、AST、面密度水平均高于正常组,差异均有统计学意义(P<0.05);模型组第4周和第7周总蛋白(TP)、肝/体质量比水平均低于正常组,差异均有统计学意义(P<0.05);两组第4周与第7周肝/体质量组内比较,差异均无统计学意义(P=0.11);模型组第4周和第7周光镜下可见ED1、α-SMA阳性细胞数量较多,主要分布于增生的纤维组织及纤维间隔弥漫部位。结论:DMN可导致大鼠肝功能障碍及弥漫肝硬化。大鼠肝纤维化模型的库普弗细胞可激活、活化肝星状细胞。
Objective:To investigate the distribution and significance of Kupffer cells and hepatic stellate cells in rat hepatic fibrosis induced by dimethylnitrosamine(DMN).Method:40 Wistar male rats were intraperitoneally injected with 10 g/L DMN for 4 weeks to prepare the hepatic fibrosis model.All rats were divided into model group(n=20)and normal group(n=20)according to the random number table method.DMN was used to establish liver fibrosis model in both groups.The levels of ALT and AST in two groups were measured by Reiman method at the 4 th and 7 th week.The distribution of ED1 and alpha-SMA markers in Kupffer cells and hepatic stellate cells of rats were determined by immunohistochemistry.The liver/body mass ratio,pathological changes of liver tissue and surface density of collagen fibers in two groups were collected and analyzed.Result:The levels of ALT,AST and area density in the model group were higher than those in the normal group at the 4 th and 7 th week(P<0.05).The levels of total protein(TP)and liver/body mass ratio in the model group were lower than those in the normal group at the 4 th and 7 th week(P<0.05).There was no significant difference in liver/body mass time between the two groups at the 4 th and 7 th week(P=0.11).The number of ED1 and α-SMA positive cells in the model group were higher at the 4 th and 7 th week under light microscope.These were mainly distributed in the proliferative fibrous tissue and the diffuse part of the fibrous septum.Conclusion:DMN can cause liver dysfunction and diffuse cirrhosis in rats.Kupffer cells in rat hepatic fibrosis model can activate and activate hepatic stellate cells.
Objective:To investigate the distribution and significance of Kupffer cells and hepatic stellate cells in rat hepatic fibrosis induced by dimethylnitrosamine(DMN).Method:40 Wistar male rats were intraperitoneally injected with 10 g/L DMN for 4 weeks to prepare the hepatic fibrosis model.All rats were divided into model group(n=20)and normal group(n=20)according to the random number table method.DMN was used to establish liver fibrosis model in both groups.The levels of ALT and AST in two groups were measured by Reiman method at the 4 th and 7 th week.The distribution of ED1 and alpha-SMA markers in Kupffer cells and hepatic stellate cells of rats were determined by immunohistochemistry.The liver/body mass ratio,pathological changes of liver tissue and surface density of collagen fibers in two groups were collected and analyzed.Result:The levels of ALT,AST and area density in the model group were higher than those in the normal group at the 4 th and 7 th week(P<0.05).The levels of total protein(TP)and liver/body mass ratio in the model group were lower than those in the normal group at the 4 th and 7 th week(P<0.05).There was no significant difference in liver/body mass time between the two groups at the 4 th and 7 th week(P=0.11).The number of ED1 and α-SMA positive cells in the model group were higher at the 4 th and 7 th week under light microscope.These were mainly distributed in the proliferative fibrous tissue and the diffuse part of the fibrous septum.Conclusion:DMN can cause liver dysfunction and diffuse cirrhosis in rats.Kupffer cells in rat hepatic fibrosis model can activate and activate hepatic stellate cells.
引文
[1]陈宵瑜,杨长青.肝纤维化发生机制研究新进展[J].实用肝脏病杂志,2016,19(1):121-124.
[2]刘明颖,杨加彩,王英杰.体内抑制库普弗细胞对D-氨基半乳糖所致大鼠肝损伤的影响[J].胃肠病学和肝病学杂志,2016,25(9):1002-1004.
[3]张伟,贾继东.肝纤维化的发病机制及治疗新靶点[J].临床肝胆病杂志,2017,33(3):409-412.
[4]Matsuda Y,Matsumoto K,Yamada A,et al.Preventive and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis[J].Hepatology,1997,26(1):81-89.
[5]李慧,汪根树.肝硬化肝移植术后脾功能亢进的研究进展[J].器官移植,2016,7(3):238-240.
[6]Zoubek M E,Trautwein C,Strnad P.Reversal of liver fibrosis:From fiction to reality[J].Best Pract Res Clin Gastroenterol,2017,31(2):129-141.
[7]严栋梁,陈杰,葛创,等.川芎嗪对刀豆蛋白A诱导的小鼠肝纤维化TNF-α、IL-6、IL-10表达的影响[J].中国医学创新,2017,14(19):19-22.
[8]滕杨,罗时旋,郭英雪,等.代谢组学法考察金银花醇提物对DMN诱导大鼠肝损伤的保护作用[J].食品研究与开发,2016,37(4):29-34.
[9]王永宏,赵晨曦,陈本美,等.茵陈蒿汤对二甲基亚硝胺诱导大鼠肝纤维化的逆转作用[J].中国中药杂志,2014,39(8):1473-1478.
[10]尹燕,李校天,郭永泽,等.1,25-二羟维生素D3对肝纤维化大鼠肝组织HIF-1α和TREM-1表达的影响[J].实用肝脏病杂志,2017,20(2):148-152.
[11]Park H J,Kim H G,Wang J H,et al.Comparison of TGF-β,PDGF,and CTGF in hepatic fibrosis models using DMN,CCl4,and TAA.[J].Drug and Chemical Toxicology,2016,39(1):111-118.
[12]Walton J C.Functionalised Oximes:Emergent Precursors for Carbon-,Nitrogen-and Oxygen-Centred Radicals[J].Molecules,2016,21(1):63.
[13]Hong I H,Park S J,Goo M J,et al.JNK1 and JNK2 regulateα-SMA in hepatic stellate cells during CCl4-induced fibrosis in the rat liver[J].Pathology International,2014,63(10):483-491.
[14]邓文升,顾磊,周鸿,等.1,25(OH)-2D-3抑制肝纤维化大鼠肝组织TGF-β1和α-SMA表达的研究[J].肝胆胰外科杂志,2015,27(3):214-218.
[15]吴芙蓉,姜玲,何晓丽,等.橙皮苷对化学性肝纤维化大鼠α-SM A表达的影响[J].安徽医药,2015,19(12):2267-2271.
[16]Li X,Peng J,Sun Z,et al.Chinese medicine CGA formula ameliorates DMN-induced liver fibrosis in rats via inhibiting MMP2/9,TIMP1/2 and the TGF-β/Smad signaling pathways[J].Acta Pharmacologica Sinica,2016,37(6):783-793.
[17]王咪咪,王艳红.胚胎来源巨噬细胞的起源及其在肝脏中功能的研究进展[J].中国临床医学,2018,25(1):123-128.
[18]Bekki Y,Yoshizumi T,Shimoda S,et al.Hepatic stellate cells secrete WFA+-M2BP:Its role in biological interactions with Kupffer cells[J].Journal of Gastroenterology and Hepatology,2017,32(7):1387-1393.
[19]Hasan I H,El-Desouky M A,Hozayen W G,et al.Protective Effect of Zingiber Officinale against CCl4-Induced Liver Fibrosis Is Mediated through Downregulating the TGF-β1/Smad3 and NF-κB/IκB Pathways[J].Pharmacology,2016,97(1-2):1-9.
[20]Lan L,Liu R,Qin L Y,et al.Transplantation of bone marrowderived endothelial progenitor cells and hepatocyte stem cells from liver fibrosis rats ameliorates liver fibrosis[J].World Journal of Gastroenterology,2018,24(2):237-247.
[2]刘明颖,杨加彩,王英杰.体内抑制库普弗细胞对D-氨基半乳糖所致大鼠肝损伤的影响[J].胃肠病学和肝病学杂志,2016,25(9):1002-1004.
[3]张伟,贾继东.肝纤维化的发病机制及治疗新靶点[J].临床肝胆病杂志,2017,33(3):409-412.
[4]Matsuda Y,Matsumoto K,Yamada A,et al.Preventive and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis[J].Hepatology,1997,26(1):81-89.
[5]李慧,汪根树.肝硬化肝移植术后脾功能亢进的研究进展[J].器官移植,2016,7(3):238-240.
[6]Zoubek M E,Trautwein C,Strnad P.Reversal of liver fibrosis:From fiction to reality[J].Best Pract Res Clin Gastroenterol,2017,31(2):129-141.
[7]严栋梁,陈杰,葛创,等.川芎嗪对刀豆蛋白A诱导的小鼠肝纤维化TNF-α、IL-6、IL-10表达的影响[J].中国医学创新,2017,14(19):19-22.
[8]滕杨,罗时旋,郭英雪,等.代谢组学法考察金银花醇提物对DMN诱导大鼠肝损伤的保护作用[J].食品研究与开发,2016,37(4):29-34.
[9]王永宏,赵晨曦,陈本美,等.茵陈蒿汤对二甲基亚硝胺诱导大鼠肝纤维化的逆转作用[J].中国中药杂志,2014,39(8):1473-1478.
[10]尹燕,李校天,郭永泽,等.1,25-二羟维生素D3对肝纤维化大鼠肝组织HIF-1α和TREM-1表达的影响[J].实用肝脏病杂志,2017,20(2):148-152.
[11]Park H J,Kim H G,Wang J H,et al.Comparison of TGF-β,PDGF,and CTGF in hepatic fibrosis models using DMN,CCl4,and TAA.[J].Drug and Chemical Toxicology,2016,39(1):111-118.
[12]Walton J C.Functionalised Oximes:Emergent Precursors for Carbon-,Nitrogen-and Oxygen-Centred Radicals[J].Molecules,2016,21(1):63.
[13]Hong I H,Park S J,Goo M J,et al.JNK1 and JNK2 regulateα-SMA in hepatic stellate cells during CCl4-induced fibrosis in the rat liver[J].Pathology International,2014,63(10):483-491.
[14]邓文升,顾磊,周鸿,等.1,25(OH)-2D-3抑制肝纤维化大鼠肝组织TGF-β1和α-SMA表达的研究[J].肝胆胰外科杂志,2015,27(3):214-218.
[15]吴芙蓉,姜玲,何晓丽,等.橙皮苷对化学性肝纤维化大鼠α-SM A表达的影响[J].安徽医药,2015,19(12):2267-2271.
[16]Li X,Peng J,Sun Z,et al.Chinese medicine CGA formula ameliorates DMN-induced liver fibrosis in rats via inhibiting MMP2/9,TIMP1/2 and the TGF-β/Smad signaling pathways[J].Acta Pharmacologica Sinica,2016,37(6):783-793.
[17]王咪咪,王艳红.胚胎来源巨噬细胞的起源及其在肝脏中功能的研究进展[J].中国临床医学,2018,25(1):123-128.
[18]Bekki Y,Yoshizumi T,Shimoda S,et al.Hepatic stellate cells secrete WFA+-M2BP:Its role in biological interactions with Kupffer cells[J].Journal of Gastroenterology and Hepatology,2017,32(7):1387-1393.
[19]Hasan I H,El-Desouky M A,Hozayen W G,et al.Protective Effect of Zingiber Officinale against CCl4-Induced Liver Fibrosis Is Mediated through Downregulating the TGF-β1/Smad3 and NF-κB/IκB Pathways[J].Pharmacology,2016,97(1-2):1-9.
[20]Lan L,Liu R,Qin L Y,et al.Transplantation of bone marrowderived endothelial progenitor cells and hepatocyte stem cells from liver fibrosis rats ameliorates liver fibrosis[J].World Journal of Gastroenterology,2018,24(2):237-247.