化浊抗纤保肝汤调控PI3K/AKT/mTOR途径抑制肝纤维化的机制研究

Study on the mechanism of Huazhuo Kangxian Baogan Decoction regulating PI3K/AKT/mTOR pathway to inhibit liver fibrosis

关键词:  肝纤维化  化浊抗纤保肝汤  成纤维细胞  磷脂肌醇3-激酶/蛋白激酶B/雷帕霉素靶蛋白

Key words:  liver fibrosis  Huazhuo Kangxian Baogan Decoction  fibroblasts  PI3K/AKT/mTOR

摘要:

[目的] 探究化浊抗纤保肝汤（HZKXD）调控磷脂肌醇3-激酶（PI3K）/蛋白激酶B（AKT）/雷帕霉素靶蛋白（mTOR）途径抑制肝纤维化的机制。[方法] 80只大鼠随机分为4组：对照组、模型组、低剂量HZKXD组和高剂量HZKXD组（每组20只）。四氯化碳（10%）腹腔注射构建纤维化模型，低剂量HZKXD组和高剂量HZKXD组灌胃剂量分别为5 mL/kg和10 mL/kg。比较各组大鼠外周血中肝功能指标、肝组织损伤、肝纤维化情况、成纤维细胞标志蛋白α-平滑肌肌动蛋白（α-SMA）水平，以及PI3K、AKT、mTOR mRNA和蛋白表达量。[结果] 4组大鼠上述指标比较差异有统计学意义（P<0.05）。模型组的谷丙转氨酶（ALT）（127.26±14.03） U/L、谷草转氨酶（AST）（260.15±27.57） U/L、肝组织损伤情况、胶原蛋白的体积分数（24.27±1.96）%、α-SMA蛋白、PI3K、AKT和mTOR mRNA和蛋白表达量高于对照组，差异有统计学意义（P<0.05）。低剂量HZKXD组的ALT（92.85±9.84） U/L、AST（198.67±21.91） U/L、肝组织损伤情况、胶原蛋白的体积分数（14.81±1.04）%、α-SMA蛋白、PI3K、AKT和mTOR mRNA和蛋白表达量显著低于模型组（P<0.05）。高剂量HZKXD组的ALT（56.44±6.35） U/L、AST（137.23±14.06） U/L、肝组织损伤情况、胶原蛋白的体积分数（7.05±0.85）%、α-SMA蛋白、PI3K、AKT和mTOR mRNA和蛋白低于模型组和低剂量HZKXD组，差异有统计学意义（P<0.05）。[结论] HZKXD可缓解肝纤维化模型大鼠肝组织中胶原蛋白累积，还可以抑制PI3K/AKT/mTOR通路减少成纤维细胞。

Abstract:

[Objective] To explore the mechanism of Huazhuo Kangxian Baogan Decoction(HZKXD) regulating PI3K/AKT/mTOR pathway to inhibit liver fibrosis.[Methods] The 80 rats were randomly divided into 4 groups:control group,model group,low-dose HZKXD group and high-dose HZKXD group (n=20). Intraperitoneal injection of carbon tetrachloride (10%) was used to construct a fibrosis model after hepatitis. The intragastric doses of the low-dose HZKXD group and the high-dose HZKXD group were 5 mL/kg and 10 mL/kg,respectively. Peripheral blood liver function indexes,liver tissue damage,liver fibrosis,fibroblast marker protein alpha smooth muscle actin (α-SMA) levels in peripheral blood of rats in each group were compared,as well as PI3K,AKT,mTOR mRNA and protein expressions.[Results] The above indicators of the four groups of rats were significantly different (P<0.05). Model group ALT (127.26±14.03) U/L,AST (260.15±27.57) U/L,liver tissue damage,collagen volume fraction (24.27±1.96)%,α-SMA protein,PI3K,AKT,mTOR mRNA and protein of the control group were significantly higher than those of the control group (P<0.05). ALT (92.85±9.84) U/L,AST (198.67±21.91) U/L,liver tissue damage,collagen volume fraction (14.81±1.04)%,α-SMA protein,PI3K,AKT,mTOR mRNA and protein expressions in the low-dose HZKXD group were significantly lower than those in the model group (P<0.05). ALT (56.44±6.35) U/L,AST (137.23±14.06) U/L,liver tissue damage,collagen volume fraction (7.05±0.85)%,α-SMA protein,PI3K,AKT and mTOR mRNA and protein in the high-dose HZKXD group were significantly lower than those in the model group and low-dose HZKXD group (P<0.05).[Conclusion] HZKXD can alleviate the accumulation of collagen in the liver tissue of liver fibrosis model rats,and can also inhibit the PI3K/AKT/mTOR pathway to reduce fibroblasts.