化滞柔肝颗粒抑制内质网应激改善非酒精性脂肪性肝炎的作用机制研究  

Study on the mechanism of Huazhi Rougan Granules in alleviating non-alcoholic steatohepatitis by inhibiting endoplasmic reticulum stress

作　　者：操颖 陈泌链 余思雨 张莉 季光 CAO Ying;CHEN Milian;YU Siyu;ZHANG Li;JI Guang(Longhua Hospital Shanghai University of Traditional Chinese Medicine/lnstitute of Digestive Diseases,Shanghai 200032,China;National Key Laboratory Project for the Integration and Innovation of Classic Prescriptions and Modern Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院/脾胃病研究所,上海200032 [2]经方与现代中药融合创新全国重点实验室,上海200032

出　　处：《中华中医药杂志》2024年第5期2195-2203,共9页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82174250)。

摘　　要：目的:探究中药复方化滞柔肝颗粒改善非酒精性脂肪性肝炎(NASH)的作用机制。方法:将雄性C57BL/6J小鼠随机分为对照组,模型组,化滞柔肝颗粒低、高剂量组,以蛋氨酸胆碱缺乏(MCD)饮食诱导NASH模型并予化滞柔肝颗粒干预;检测小鼠体质量、肝重、肝体比、血清肝酶[丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)]、肝脂[总胆固醇(TC)、甘油三酯(TG)]及血清炎症因子[白细胞介素(IL)-6、IL-1β、肿瘤坏死因子α(TNF-α)]水平,结合肝组织病理染色结果评估化滞柔肝颗粒对NASH小鼠表型的影响;通过RNAseq、RT-qPCR、Western Blot、ELISA和TUNEL染色等方法探究化滞柔肝颗粒调节内质网应激(ERS)与细胞凋亡改善NASH的效应机制。结果:化滞柔肝颗粒可降低NASH小鼠血清ALT、AST,肝脏TC、TG和血清炎症因子水平(P<0.01,P<0.05),减轻肝脏脂肪变和炎症浸润,下调ERS相关基因和蛋白的表达;其有效成分大黄素能降低Raw264.7细胞的炎症因子水平,下调Bcl-2/Bax/Caspase3信号通路基因和蛋白的表达,抑制细胞凋亡。结论:化滞柔肝颗粒能有效改善NASH,其机制可能与通过Bcl-2/Bax/Caspase3信号通路抑制巨噬细胞凋亡,改善ERS有关。Objective:To explore the mechanism of Huazhi Rougan Granules(HZRG)in alleviating non-alcoholic steatohepatitis(NASH).Methods:Male C57BL/6J mice were randomly divided into control group,model group,HZRG low/high-dose group.The NASH model was induced by methionine choline deficiency(MCD)diet with the intervention of HZRG.The body weight,liver weight,liver body ratio,serum liver enzymes(AST,ALT),liver fat(TC,TG)and serum inflammatory factors(IL-6,IL-1β,TNF-α)of mice were detected.The efficacy of HZRG was evaluated by pathological staining and immunohistochemical staining.The mechanism of HZRG regulating endoplasmic reticulum stress and apoptosis in ameliorating NASH was investigated by RNA-seq,RT-qPCR,WB,ELISA and TUNEL staining.Results:HZRG could decrease the levels of serum AST,ALT,TC,TG and serum inflammatory factors in NASH mice(P<0.01,P<0.05),alleviate liver steatopathy and inflammatory infiltration,and down-regulate the expression of ER stress-related genes and proteins.Emodin,the active component of HZRG,can significantly reduce the level of inflammatory factors in Raw264.7 cells,down-regulate the expression of Bcl-2/Bax/Caspase3 pathway genes and proteins,and inhibit cell apoptosis.Conclusion:HZRG can effectively ameliorate NASH,and its mechanism may be related to inhibiting macrophage apoptosis and ameliorating ER stress through Bcl-2/Bax/Caspase3 signaling pathway.

关 键 词：化滞柔肝颗粒 非酒精性脂肪性肝炎 内质网应激 Bcl-2/Bax/Caspase3信号通路 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]