摘要
目的探讨柔肝降酶方对肝纤维化大鼠的作用机制。方法将大鼠分为正常组、模型组、秋水仙碱组(0.2 mg/kg)和柔肝降酶方低、高剂量组(9.9、19.8 g/kg),每组10只,采用腹腔注射橄榄油CCl_(4)混合溶液(40%,3 mL/kg)制备肝纤维化大鼠模型,灌胃给药6周后,检测血清ALT、AST水平,ELISA法检测血清TNF-α、IL-6水平,免疫组化法(IHC)检测肝组织α-SMA表达,Western blot法检测肝组织PI3K/Akt/mTOR信号通路相关蛋白(PI3K、p-PI3K、Akt、p-Akt、mTOR、p-mTOR)、自噬相关蛋白(P62、Beclin-1、LC3-Ⅰ、LC3-Ⅱ)、纤维化相关蛋白(α-SMA、Col-1)的表达。结果与正常组比较,模型组大鼠血清ALT、AST、TNF-α、IL-6水平、肝组织α-SMA阳性细胞表达及α-SMA、Col-1、Beclin-1、LC3-Ⅱ蛋白表达均升高(P<0.01),肝组织P62、p-PI3K、p-Akt、p-mTOR蛋白表达降低(P<0.05,P<0.01);与模型组比较,柔肝降酶方各剂量组大鼠血清ALT、AST、TNF-α、IL-6水平、肝组织α-SMA阳性细胞表达及α-SMA、Col-1、Beclin-1、LC3-Ⅱ蛋白表达降低(P<0.01),肝组织P62、p-PI3K、p-Akt、p-mTOR蛋白表达升高(P<0.05,P<0.01)。结论柔肝降酶方可能通过调控PI3K/Akt/mTOR信号通路抑制自噬,起到抗CCl_(4)诱导的大鼠肝纤维化作用。
AIM To investigate the effects and mechanism of Rougan Jiangmei Recipe on hepatic fibrosis in rats.METHODS The rats were divided into the normal group,the model group,colchicine group(0.2 mg/kg)and Rougan Jiangmei Recipe group(9.9 g/kg low dose use and 19.8 g/kg high dose use,respectively),with 10 rats in each group.The rat model of hepatic fibrosis was made by intraperitoneal injection of olive oil CCl_(4)mixed solution(40%,3 mL/kg).After 6 weeks of corresponding intragastric administration of the agent,the rats had their serum ALT and AST levels detected;their serum TNF-α and IL-6 levels detected by ELISA;their hepatic α-SMA expression detected by IHC;and their hepatic PI3K/Akt/mTOR signaling pathway related proteins(PI3K,p-PI3K,Akt,p-Akt,mTOR,p-mTOR),autophagy related proteins(P62,Beclin-1,LC3-Ⅰ,LC3-Ⅱ)and fibrosis related proteins(α-SMA,Col-1)determined by Western blot.RESULTS Compared with the normal group,the model group displayed increased levels of serum ALT,AST,TNF-α,IL-6,and hepatic α-SMA positive cells expression,α-SMA,Col-1,Beclin-1,and LC3-Ⅱ protein expressions(P<0.01),and decreased hepatic protein expressions of P62,p-PI3K,p-Akt,and p-mTOR(P<0.05,P<0.01).Compared with the model group,Rougan Jiangmei Recipe groups shared reduced serum ALT,AST,TNF-α,IL-6 levels,and hepaticα-SMA positive cells expression,α-SMA,Col-1,Beclin-1,and LC3-II protein expressions(P<0.01),and increased hepatic protein expressions of P62,p-PI3K,p-Akt,and p-mTOR(P<0.05,P<0.01).CONCLUSION Rougan Jiangmei Recipe may reverse CCl_(4)-induced liver fibrosis in rats through PI3K/Akt/mTOR signaling pathway-mediated autophagy inhibition.
作者
陶柏楠
王咏兰
田梦杰
侯孜明
万星
袁林
TAO Bo-nan;WANG Yong-lan;TIAN Meng-jie;HOU Zi-ming;WAN Xing;YUAN Lin(Hubei Provincial Key Laboratory for Interpretation and Intervention of Rheumatic Diseases,Hubei Minzu University,Enshi 445000,China;West China School of Preclinical Sciences and Forensic Medicine,Sichuan University,Chengdu 610064,China)
出处
《中成药》
CAS
CSCD
北大核心
2023年第3期741-747,共7页
Chinese Traditional Patent Medicine
基金
国家自然科学基金资助项目(81560675)
湖北省卫健委面上项目(ZY2019M076)
恩施州科技局科技计划性项目(E20190001)。
作者简介
陶柏楠(1996-),男,硕士,研究方向为中医药防治肝脏疾病。Tel:13355244267,E-mail:2373247494@qq.com;通信作者:袁林(1974-),男,副教授,从事中医药及抗纤维化研究。Tel:(0718)8301984,E-mail:171354806@qq.com。
