基于TGF-β1/Smads信号通路探究强肝软坚丸对肝纤维化大鼠肝脏的保护作用

Exploring the protective effect of Qianggan Ruanjian pill on liver fibrosis in rats based on TGF-β1/Smads signaling pathway

关键词:  强肝软坚丸  TGF-β1/Smad通路  肝纤维化  肝脏

Key words:  Qianggan Ruanjian pill  TGF-β1/Smad pathway  liver fibrosis  liver

摘要:

[目的] 基于转化生长因子-β1（TGF-β1）/Smads信号通路探究强肝软坚丸对肝纤维化（HF）大鼠肝脏的保护作用。[方法] 将Wistar大鼠分为对照组、HF组、强肝软坚丸组、秋水仙碱组、SRI-011381组、强肝软坚丸+SRI-011381组，每组12只。除对照组外，其他组大鼠均通过腹腔注射1.5 mL/kg 50%四氯化碳（CCl₄）橄榄油溶液的方式构建HF模型，建模成功后，进行给药处理，给药每日1次，持续6周。检测大鼠血清中谷丙转氨酶（ALT）、谷草转氨酶（AST）活性及Ⅳ型胶原蛋白（CIV）、Ⅲ型前胶原蛋白（PC-Ⅲ）、层粘连蛋白（LN）、透明质酸（HA）含量；苏木精-伊红（HE）、马松（Masson）染色分别检测肝脏组织病理变化、肝纤维化情况；免疫组化检测大鼠肝脏组织中α-平滑肌肌动蛋白（α-SMA）阳性细胞数；蛋白免疫印迹法（Western blot）检测肝脏组织中TGF-β1、p-Smad2/3蛋白表达。[结果] 与对照组相比，HF组大鼠肝组织病理损伤及肝纤维化严重，血清中ALT、AST活性、CIV、PC-Ⅲ、LN、HA含量、肝组织中α-SMA阳性细胞数及TGF-β1、p-Smad2/3蛋白表达升高（P＜0.05）；与HF组相比，强肝软坚丸组、秋水仙碱组大鼠肝组织病理损伤及肝纤维化有所改善，血清中ALT、AST活性、CIV、PC-Ⅲ、LN、HA含量、肝组织中α-SMA阳性细胞数及TGF-β1、p-Smad2/3蛋白表达降低，SRI-011381组对应指标变化趋势与上述相反（P＜0.05）；与强肝软坚丸组相比，强肝软坚丸+SRI-011381组大鼠肝组织病理损伤及肝纤维化加剧，血清中ALT、AST活性、CIV、PC-Ⅲ、LN、HA含量、肝组织中α-SMA阳性细胞数及TGF-β1、p-Smad2/3蛋白表达升高（P＜0.05）。[结论] 强肝软坚丸可能通过抑制TGF-β1/Smads信号通路改善大鼠HF。

Abstract:

[Objective] To investigate the protective effect of Qianggan Ruanjian pill on the liver of rats with liver fibrosis（HF） based on the growth factor-β1（TGF-β1）/Smads signaling pathway. [Methods] Wistar rats were separated into a control group，HF group，Qianggan Ruanjian pill group，colchicine group，SRI-011381 group，and Qianggan Ruanjian pill+SRI-011381 group，with 12 rats in each group. Except for the control group，rats in all other groups were injected intraperitoneally with a 50% carbon tetrachloride（CCl₄） olive oil solution of 1.5 mL/kg to construct HF models. After successful modeling，medication was administered once a day for 6 weeks. The activity of alanine aminotransferase（ALT） and aspartate aminotransferase（AST） in serum，and the contents of type Ⅳ collagen（CIV），type Ⅲ procollagen（PC-Ⅲ），laminin（LN），and hyaluronic acid（HA） were detected；hematoxylin-eosin（HE） and Masson staining were applied to detect pathological changes in liver tissue and liver fibrosis，respectively；immunohistochemistry was applied to detect the number of α-smooth muscle actin（α-SMA） positive cells in liver tissue；Western blot was applied to detect the expression of TGF-β1 and p-Smad2/3 proteins in liver tissue. [Results] ACompared with the control group，the HF group had severe liver tissue pathological damage and liver fibrosis，the activities of ALT and AST in serum，the contents of CIV，PC-Ⅲ，LN，HA，the number of α-SMA positive cells in liver tissue，and the expression of TGF-β1 and p-Smad2/3 proteins increased（P＜0.05）；compared with the HF group，the liver tissue pathological damage and liver fibrosis in the Qianggan Ruanjian pill group and the colchicine group were improved，the activities of ALT and AST in serum，the contents of CIV，PC-Ⅲ，LN，HA，the number of α-SMA positive cells in liver tissue，and the expression of TGF-β1 and p-Smad2/3 proteins decreased，the change trend of corresponding indicators in SRI-011381 group was opposite to the above（P＜0.05）；compared with the Qianggan Ruanjian pill group，the pathological damage and liver fibrosis of the liver tissue in the Qianggan Ruanjian pill+SRI-011381 group were exacerbated，the activities of ALT and AST in serum，the contents of CIV，PC-Ⅲ，LN，HA，the number of α-SMA positive cells in liver tissue，and the expression of TGF-β1 and p-Smad2/3 proteins increased（P＜0.05）. [Conclusions] Qianggan Ruanjian pill may improve HF in rats by inhibiting the TGF-β1/Smads signaling pathway.