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2020, v.45(19) 4725-4731

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竹节参皂苷Ⅳa调节miR-17-5p/MFN2信号通路改善非酒精性脂肪性肝炎
Effect of Panax japonicus saponin Ⅳa on alleviating nonalcoholic steatohepatitis by regulating miR-17-5p/MFN2 signaling pathway

李聪;刘小慧;熊海容;李晓晓;罗悦;刘朝奇;
LI Cong;LIU Xiao-hui;XIONG Hai-rong;LI Xiao-xiao;LUO Yue;LIU Chao-qi;Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy, China Three Gorges University;Medical College of China Three Gorges University;

摘要(Abstract):

该文研究了竹节参皂苷Ⅳa(Panax japonicus saponinⅣa, SPJ-Ⅳa)通过miR-17-5p/mitofusin 2 (MFN2)信号通路改善非酒精性脂肪性肝炎(nonalcoholic steatohepatitis, NASH)的作用。采用高脂饮食(high-fat diet,HFD)联合CCl_4诱导Balb/c雄性小鼠NASH模型,并采用不同剂量SPJ-Ⅳa进行干预,处理5周后留取血清及肝脏,称量体质量及肝重,计算肝指数,检测血清生化指标丙氨酸氨基转移酶(ALT)、甘油三酯(TG)及葡萄糖(Glu)的含量,HE及Masson染色法观察肝脏形态学变化及纤维化病变情况,Real-time PCR法检测脂代谢相关基因[脂肪酸合成基因(fatty acid synthase gene,FASN)、碳水化合物反应元件结合蛋白(carbohydrate response element binding protein,chREBP)、乙酰辅酶A羧化酶1 (acetyl CoA carboxylase 1,ACC1)、肝X受体(liver X receptor,LXR)]、炎症因子白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)、miR-17-5p及MFN2的表达情况,Wes-tern blot法检测MFN2蛋白表达水平。结果显示,与正常组相比,HFD+CCl_4组中肝指数明显增加,生化指标ALT,TG,Glu的含量明显增高;形态学结果显示有大量脂肪空泡出现,表现出明显的脂肪变性,并可见明显的胶原纤维沉积;脂代谢相关基因、炎症因子IL-6和IL-1β及miR-17-5p mRNA表达水平明显增加,MFN2 mRNA表达水平明显降低;MFN2蛋白水平降低。经SPJ-Ⅳa干预后,生化指标ALT,TG,Glu的含量有所下降,脂肪变性有所减轻,胶原纤维沉积有所减少,脂代谢相关基因、炎症因子IL-6和IL-1β及miR-17-5p mRNA表达水平有所下降,MFN2 mRNA表达水平有所升高;MFN2蛋白水平也升高。该研究结果表明,miR-17-5p/MFN2信号通路可能参与了NASH的发生发展,且SPJ-Ⅳa对NASH有保护作用,其机制可能与调控miR-17-5p/MFN2信号通路有关。
To study the effect of Panax japonicas saponin Ⅳa(SPJ-Ⅳa) on nonalcoholic steatohepatitis(NASH) through miR-17-5 p/MFN2 signaling pathway. The nonalcoholic steatohepatitis model was induced by a high-fat diet combined with CCl_4 in Balb/c male mice. The mouse serum and liver were collected, the body weight and liver weight were measured, the liver index was calculated, and the serum biochemical indicators alanine amino transferase(ALT), triglyceride(TG), and glucose(Glu) were measured. The morphological changes in the liver were detected by HE and Masson staining, Real-time PCR was used to detect lipid metabolism-related genes, inflammation-related genes interleukin-6(IL-6) and interleukin-1β(IL-1β), miR-17-5 p and MFN2 expressions, and Western blot was used to detect MFN2 protein expression level. Compared with the normal control group, the liver index in the HFD+CCl_4 group was significantly increased, and the contents of ALT, TG, and Glu were significantly increased; the morphology showed obvious steatosis and collagen fiber deposition; mRNA expression levels of lipid metabolism-related genes, inflammation-related genes and miR-17-5 p increased significantly, the mRNA expression level of MFN2 decreased significantly, and the protein level of MFN2 decreased. After intervention with SPJ-Ⅳa, the levels of ALT, TG and Glu decreased, morphological steatosis decreased, collagen fiber deposition decreased, and mRNA expression levels of lipid metabolism-related genes, inflammation-related genes and miR-17-5 p decreased. The mRNA expression level of MFN2 increased, and the protein level of MFN2 also increased. The results of this study indicated that miR-17-5 p/MFN2 signaling pathway may be involved in the occurrence and development of NASH, and SPJ-Ⅳa had a protective effect on NASH, its mechanism may be related to the regulation of miR-17-5 p/MFN2 signaling pathway.

关键词(KeyWords): 非酒精性脂肪性肝炎;CCl_4;竹节参皂苷Ⅳa;MFN2;miR-17-5p
nonalcoholic steatohepatitis;CCl_4;Panax japonicus saponin Ⅳa;MFN2;miR-17-5p

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81673675);; 三峡大学学位论文培优基金项目(2019SSPY102)

作者(Author): 李聪;刘小慧;熊海容;李晓晓;罗悦;刘朝奇;
LI Cong;LIU Xiao-hui;XIONG Hai-rong;LI Xiao-xiao;LUO Yue;LIU Chao-qi;Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy, China Three Gorges University;Medical College of China Three Gorges University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20200427.401

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