双氢青蒿素对胆管结扎致大鼠肝纤维化的治疗作用研究

Therapeutical Effect of Dihydroartemisinin on Bile Duct Ligation-Induced Liver Fibrosis in Rats and Its Mechanism

目的探究双氢青蒿素(Dihydroartemisinin,DHA)对胆管结扎(Bile duct ligation,BDL)所致的大鼠肝纤维化的初步治疗作用。方法采用BDL诱导大鼠肝纤维化模型,DHA(14mg/kg)及阳性药物秋水仙碱(0.1mg/kg)干预处理,末次给药24h后,取血清检测ALT、AST、Bilirubin;HE染色、马松染色和天狼猩红染色检测肝组织病理变化;Western blot检测肝组织中肝星状细胞(HSC)活化指标α-SMA与α1(Ⅰ)collagen的表达;免疫荧光检测肝组织TGFβ-RⅡ的表达。体外培养活化的HSC-T6,并用DHA干预处理,Western blot检测α-SMA、α1(Ⅰ)collagen以及TGFβ-RⅡ、p-Smad2、Smad2的表达。结果血清学指标结果显示,与空白对照组比较,模型组SD大鼠ALT、AST、Bilirubin显著升高,DHA能降低血清中ALT、AST、Bilirubin水平;HE、马松染色、天狼猩红染色结果显示,与模型组比较,DHA能够明显改善肝脏病理组织结构以及胶原纤维的沉积;Western blot结果显示,DHA还可抑制HSC活化的标志物α-SMA、α1(Ⅰ)collagen以及TGFβ-RⅡ、p-Smad2的表达。结论 DHA对BDL诱导的肝纤维化具有显著的保护作用,这一效应主要与其抑制HSC活化以及调控TGF-βⅡ/Smad2信号通路相关。

OBJECTIVE To investigate the therapeutical effect of dihydroartemisinin on bile duct ligation-induced liver fibro sis in rats. METHODS Hepatic fibrosis model was established by bile duct ligation, treatment groups in which rats received intraperitoneal injection of DHA at 14 mg/kg and colehicine treatment. At the end of the experiment, the rats were sacrificed, Blood was collected to exam Levels of alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, and Bilirubin in serum samples. Hematoxylin and eosin （HE）, Masson＇ s triehrome stain and sirius red collagen staining were used to Patho- logical changes in liver tissue. Western blotting analyses were used to evaluate the indicators related with HSC activation such as a-SMA, α 1 （ I ） collagen in liver tissue. Immunofluoreseence was used to evaluate the expression of TGFβ-R Ⅱ in liver tis- sue. HSC-T6 was cultured in vitro and treated with DHA. Western blotting analyses were used to evaluate the expressions of a-SMA, a 1（ I ） collagen, TGFβ-RⅡ , p-Smad2, and Smad2. RESULTS As shown in Serological indicators, compared with the control group, contents of serum ALT, AST, Bilirubin was significantly increased in the model group, DHA could de- crease the serum levels of ALT, AST, Bilirubin. As shown in HE, Masson, and Sirius red staining, comparedwith the model group, DHA significantly alleviated liver pathology tissue and deposition of collagen fibers, and inhibited expression of a-SMA, a 1（ [ ） collagen and TGFβ-RⅡ , p-Smad2. CONCLUSION DHA had the protective effect on liver fibrosis induced by BDL, which was primarily related to inhibition of HSC activation and regulation of TGF -βⅡ/Smad2 signaling pathways.