摘要
目的 :探讨miR-378对非酒精性脂肪性肝炎(NASH)发生及发展的作用及其机制。方法 :将C57BL/6小鼠随机分为对照(正常饮食)组、NASH模型(高脂饮食)组、miR-378抑制剂(高脂饮食+antagomir-378)组、阴性抑制剂(高脂饮食+antagomir阴性对照)组。观察小鼠肝系数和组织病理改变;采用实时荧光定量PCR(qRT-PCR)验证NASH小鼠及各处理组肝组织的miR-378表达。应用自动化学分析仪检测血清丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)的水平;qRT-PCR检测肝组织肿瘤坏死因子-α(TNF-α)、白细胞介素-4(IL-4)、白细胞介素-6(IL-6)、单核细胞趋化蛋白-1(MCP-1)、转化生长因子β(TGF-β)和胶原蛋白1α2(Col1α2)的mRNA表达;免疫荧光检测TGF-β和Col1α2蛋白表达。结果 :与对照组比较,NASH模型组和阴性抑制剂组的肝系数显著升高;与NASH模型组比较,miR-378抑制剂组肝系数显著下降。NASH模型组肝小叶结构不清,出现明显的肝细胞坏死、脂肪及炎症细胞浸润;miR-378抑制剂组肝小叶结构正常,肝细胞排列整齐,脂肪浸润减轻。NASH模型组和阴性抑制剂组miR-378表达较对照组显著升高;NASH模型组和阴性抑制剂组血清ALT和AST含量较对照组显著升高,而miR-378抑制剂组较NASH模型组含量显著下降;NASH模型组和阴性抑制剂组TNF-α、IL-6表达量较对照组明显升高,而IL-4明显下降,MCP-1差异无统计学意义;而miR-378抑制剂组TNF-α、IL-6表达量较NASH模型组明显下降,IL-4明显上升;NASH模型组和阴性抑制剂组TGF-β和Col1α2表达量较对照组明显升高;而与NASH模型组比较,miR-378抑制剂组TGF-β和Col1α2表达量明显下降。结论 :miR-378可能是NASH的危险因素,抑制miR-378的表达有利于缩短NASH病程,为NASH治疗提供了新的潜在治疗靶点。
Objective:To study the role and mechanism of miR-378 in the occurrence and development of non-alcoholic steatohepatitis(NASH)animal model.Methods:C57BL/6 mice were randomly divided into control(normal diet)group,NASH model(high fat diet)group,miR-378 inhibitor group(high fat diet+antagomir-378)group,negative inhibitor(high-fat diet+antagomir negative control)group.Liver coefficient and histopathological changes was observed in mice.The real-time quantitative PCR(qRT-PCR)was used to verify the expression of miR-378 in liver tissues of mice with NASH disease.The levels of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured by Olympus automatic chemical analyzer.The qRT-PCR was used to detect the expression levels of tumor necrosis factor-α(TNF-α),interleukin 4(IL-4),interleukin 6(IL-6),monocyte chemoattractant protein-1(MCP-1)and transforming growth factor-β(TGF-β)and collagen type lα2(Col1α2)in liver tissues.Results:Compared with the control group,the liver coefficients of NASH model group and negative inhibitor group were significantly increased.Compared with the NASH model group,the liver coefficient of miR-378 inhibitor group was decreased.The structure of hepatic lobule in the NASH model group was unclear,and there were obvious hepatocyte necrosis,hepatic cord disorder and inflammatory cell infiltration.Then in the miR-378 inhibitor group,the structure of the hepatic lobule was normal under light microscope,and the hepatocytes were neatly arranged,and the fat infitration was allevuated.The expression of miR-378 in the NASH model group and in the negative inhibitor group was significantly higher than that in the control group.Serum ALT and AST levels were significantly higher in NASH model group and in the negative inhibitor group than those in the control group.Compared with the NASH model group,miR-378 inhibitor group showed a significant decrease in the serum ALT and AST levels.The expression of TNF-αand IL-6 in the NASH model group and in the negative inhibitor group were significantly higher than those in the control group,while IL-4 was significantly decreased,and the difference in MCP-1 was not statistically significant.Compared with the NASH model group and in the negative inhibitor group,the expression of TNF-αand IL-6 in the miR-378 inhibitor group was decreased significantly and IL-4 was increased significantly.The expression of TGF-βand Col1α2 was significantly higher in the NASH model group and the negative inhibitor group than those in the control group.In comparison to the NASH model group,the miR-378 inhibitor group showed a significant decrease in the expression levels of TGF-βand Col1α2.Conclusion:MiR-378 may be a risk factor for NASH.Therefore,inhibiting the expression of miR-378 is beneficial for shortening the course and treatment of NASH disease,providing a new potential therapeutic target for NASH.
作者
佘明豪
杨文辉
刘寒松
余洋
张磊
She Minghao;Yang Wenhui;Liu Hansong;Yu Yang;Zhang Lei(Department of General Surgery,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,China;Department of Gastroenterology,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,China)
出处
《解剖学杂志》
CAS
2023年第5期385-389,431,共6页
Chinese Journal of Anatomy
基金
河南省高等学校重点科研项目计划(20A320052)。
