摘要
目的:观察甘草酸二铵对继发性胆汁淤积性大鼠肝纤维化形成中肝细胞向肌成纤维细胞转分化(EMT)的影响。方法:75只SD雄性大鼠采用随机分为假手术对照组(Sham,n=15)及胆汁淤积性肝纤维化模型对照组(M,n=30)及药物干预组(Y,n=30)。M及Y组采用胆总管结扎(bile duct ligation,BDL)制备胆汁淤积性肝纤维化模型,Sham组仅作胆总管分离,不作胆总管结扎。Y组于术后1周开始经尾静脉予甘草酸二铵,1日1次,共4周。所有大鼠均于术后第5周处死取材。统计分析实验过程大鼠死亡率及腹水发生率,观察实验大鼠肝功能、观测肝脏组织学及羟脯氨酸含量(Hyp);激光共聚焦显微镜观察肝组织肝细胞特异性抗原(Hep par)及上皮α-平滑肌肌动蛋白(α-SMA)共定位,Western blot检测Hep par的、α-SMA蛋白表达。结果:与Sham组比较,M组大鼠肝功能显著异常,肝组织纤维增生程度、Hyp含量及α-SMA蛋白表达量均显著增加(P<0.01),而HepPar蛋白表达显著减少(P<0.01),并出现大量Hep par/α-SMA共染阳性细胞;与M组比较,Y组大鼠死亡率、腹水发生率显著下降(P<0.01),肝功能显著改善(P<0.01),肝组织纤维增生程度、Hyp含量及α-SMA蛋白表达量显著下降(P<0.01),HepPar蛋白表达显著升高(P<0.01)。结论:甘草酸二铵可有效地抑制BDL大鼠肝纤维化模型肝组织肝细胞向肌成纤维细胞转分化,这可能也是甘草酸二铵干预胆汁淤积性肝纤维化的主要效应机制之一。
Objective：To investigate the influence of diammonium glycyrrhizinate for hepatocytes transdifferentiate tomyofibroblast on secondary cholestatic hepatic fibrosis rats. Methods：Cholestatic hepatic fibrosis in seventy SD male rats were induced by ligation of bile duct,and sham control group（n = 15）was only made choledochus segregated. 1 week af- ter BDL, rats were divided into model control group （M, n = 30）and drugs intervention group randomly （Y, n =30 ）. The diammonium glycyrrhizinate intervention was lasted for 4 weeks by caudal vein injection, then rats of M group were NS. All of rats were sacrificed 5 weeks after operation for determining liver histology, hepatic function and hepatic hydroxypro- line content（Hyp）. Co- immunofluorescence staining of the myofibroblast marker α -SMA and the hepatocyte specific antigen Hep par were detected by laser confocal microscopy, while protein expression of Hep par and α - SMA were meas- ured by Western blot analysis. Result ： Compared with that of sham control group, mortality of model control group rats was 33.3% and ascites frequency was 90.0% , while hepatic function was abnormal significantly, Hyp content of liver tissue and protein expression of ct - SMA were increased obviously in model control group, but protein expression of Hep par was decreasely obviously, and many positive ceils of Hep Par/α - SMA were observed. Compared with that of model control group, mortality and ascites frequency were decreased significantly and weight of spleen were lessened obviously, while he- patic function was improved significantly. Hyp content of liver tissue and protein expression of α- SMA were both de- creased（P 〈0.01 ）, while protein expression of Hep par was increasely obviously. Conclusions：Diammonium glycyrrhiz- inate can restrain the hepatocytes transdifferentiate to myofibroblast, which may be a key step in the process of interventing and treating cholestasis fibrosis.
出处
《中华中医药学刊》
CAS
2013年第4期824-827,I0004,共5页
Chinese Archives of Traditional Chinese Medicine
基金
浙江省教育厅科研资助项目(Y200908063)
浙江省卫生厅科技计划资助项目(2010kyb086)
