薯蓣皂苷降低四氯化碳致大鼠肝纤维化作用及其机制的研究

Diosgenin Reduces Liver Fibrosis Induced by Carbon Tetrachloride and its Mechanism

探索薯蓣皂苷对四氯化碳(CCl)诱导的大鼠肝纤维化的保护作用,并探讨其机制。将体重180~220 g的32只雄性SD大鼠随机分为4组(每组n=8),包括对照组、薯蓣皂苷组、CCl组,以及用薯蓣皂苷治疗的CCl组。采用腹腔注射20%CCl(2.5 mL/kg体重)的橄榄油产生大鼠肝脏纤维化损伤。H&E、Masson和IHC染色观察肝脏组织学变化,胶原沉积情况,以及α-SMA的表达,并通过实时PCR将数据定量化;试剂盒法检测大鼠的AST和ALT水平;ELISA法检测大鼠血清中炎性因子的水平;免疫印迹法检测薯蓣皂苷对肝纤维化大鼠Wnt1,β-catenin、c-Myc和cyclin D1的蛋白表达的影响。结果显示,薯蓣皂苷处理可显著下调CCl诱导的大鼠肝纤维化模型中肝纤维化标记物[包括α-平滑肌肌动蛋白(α-SMA)和胶原蛋白Ⅰ]的蛋白表达,同时降低天冬氨酸转氨酶(AST)和丙氨酸转氨酶(ALT)的升高,降低促炎因子并提升抑炎因子的表达。此外,薯蓣皂苷降低肝纤维化大鼠Wnt1、β-catenin、c-Myc和cyclin D1的表达,这表明薯蓣皂苷至少可以通过抑制Wnt/β-catenin信号传导途径来减轻肝脏纤维化。薯蓣皂苷可以通过抑制Wnt/β-catenin信号通路,减轻四氯化碳诱导的大鼠肝纤维化损伤。

To explore the protective effect of diosgenin on hepatic fibrosis induced by carbon tetrachloride(CCl)and its mechanism,32 rats were randomly divided into 4 groups(n=8 in each group),including control group,diosgenin group,CClgroup,and CCl+diosgenin group.Rat liver fibrosis injury was induced by intraperitoneal injection of 20%CCl(2.5 mL/kg body weight).H&E,Masson and IHC staining were used to observe liver histological changes,collagen deposition,andα-SMA expression;And then quantify data by real-time PCR;Kits were used to detect AST and ALT levels in rats;ELISA was used to detect levels of inflammatory factors in rat serum;The expression of Wnt1,β-catenin,c-Myc and cyclin D1 in hepatic fibrosis rats were detected.The results showed that diosgenin treatment significantly down-regulated the protein expression of liver fibrosis markers[includingα-smooth muscle actin(α-SMA)and collagen I]in CCl-induced rat liver fibrosis model.At the same time,it reduces the increase of aspartate aminotransferase(AST)and alanine aminotransferase(ALT),and reduces pro-inflammatory factors and promotes the expression of anti-inflammatory factors.In addition,diosgenin reduced the expression of Wnt1,β-catenin,c-Myc and Cyclin D1 in hepatic fibrosis rats,suggesting that diosgenin can at least reduce liver fibrosis by inhibiting Wnt/β-catenin signaling pathway.Diosgenin can attenuate carbon tetrachloride-induced liver fibrosis injury in rats by inhibiting Wnt/β-catenin signaling pathway.