过表达lncRNAHEM2M改善非酒精性脂肪肝病小鼠的肝脏损伤  

Overexpression of lncRNA HEM2M alleviates liver injury in mice with non-alcoholic fatty liver disease

作　　者：孔祥[1,2,3] 张腾 张妍[1,4] 高灵犀[1,4] 汪文 汪梦燕 王国栋 吕坤 KONG Xiang;ZHANG Teng;ZHANG Yan;GAO Linxi;WANG Wen;WANG Mengyan;WANG Guodong;LÜKun(Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institution,Wannan Medical College,Wuhu 241001,China;Central Laboratory,Yijishan Hospital,Wannan Medical College,Wuhu 241001,China;Department of Endocrinology,Yijishan Hospital,Wannan Medical College,Wuhu 241001,China;Department of Gastroenterology,Yijishan Hospital,Wannan Medical College,Wuhu 241001,China;School of Pharmacy,Wannan Medical College,Anhui Provincial Engineering Research Center for Polysaccharide Drugs,Wuhu 241002,China)

机构地区：[1]皖南医学院非编码RNA基础与临床转化安徽省重点实验室,安徽芜湖241001 [2]皖南医学院弋矶山医院中心实验室,安徽芜湖241001 [3]皖南医学院弋矶山医院内分泌科,安徽芜湖241001 [4]皖南医学院弋矶山医院消化内科,安徽芜湖241001 [5]皖南医学院药学院//安徽省多糖药物工程技术研究中心,安徽芜湖241002

出　　处：《南方医科大学学报》2024年第1期1-8,共8页Journal of Southern Medical University

基　　金：国家自然科学基金(81970699,82072370);安徽省高校杰出青年科研项目(2022AH020075);弋矶山医院“高峰”培育计划项目(GF2019J04)

摘　　要：目的探讨过表达长链非编码RNA(lncRNA)HEM2M对非酒精性脂肪肝病(NAFLD)小鼠肝损伤的影响。方法野生型C57BL/6(WT)和条件性髓系细胞lncRNAHEM2M过表达(MYKI)小鼠分别喂饲普通饮食(ND)和高脂饮食(HFD),即为WT+ND、MYKI+ND、WT+HFD和MYKI+HFD组。12周后行腹腔糖耐量及胰岛素耐量试验后处死小鼠,检测小鼠血清和肝脏组织的肝功能指标,制备肝脏组织切片后行HE染色和F4/80免疫组化染色,ELISA法检测肝脏组织中IL-6、IL-1β和TNF-α水平,qRT-PCR检测M1型(TNF-α、iNOS和IL-6)和M2型(Arg-1、YM-1和IL-10)巨噬细胞标志物mRNA表达,免疫印迹检测肝脏组织中P-AKT、T-AKT、NLRC4、caspase-1和GSDMD蛋白表达,比色法和免疫荧光测定肝脏组织caspase-1活性。结果与HFD喂饲的WT小鼠相比,MYKI+HFD小鼠肝功能损伤减轻(P<0.01),肝脏脂肪变缓解,肝脏巨噬细胞浸润减少,糖耐量损伤及胰岛素抵抗改善(P<0.01);MYKI+HFD小鼠肝脏组织IL-6、IL-1β和TNF-α水平降低(P<0.01),M1型巨噬细胞标志物mRNA表达减少(P<0.01),M2型mRNA表达增加(P<0.01);MYKI+HFD小鼠肝脏组织NLRC4炎症小体活性降低(P<0.01),活性caspase-1减少,GSDMD-N蛋白表达降低(P<0.05)。结论过表达lncRNAHEM2M降低NAFLD小鼠肝脏炎症因子水平,进而改善胰岛素抵抗并抑制肝脏NLRC4炎症小体激活,减少肝细胞焦亡,最终改善NAFLD小鼠肝脏损伤。Objective To explore the effects of long non-coding RNA(lncRNA)HEM2M overexpression on liver injury in mice with non-alcoholic fatty liver disease(NAFLD).Methods Wild-type C57BL/6(WT)mice and myeloid cell-specific HEM2M knock-in(MYKI)mice were fed normal(ND)or high-fat diet(HFD)for 12 weeks.After intraperitoneal glucose tolerance and insulin tolerance tests,the mice were euthanized for detection of liver function indicators in the serum and liver tissue.HE staining and F4/80 immunohistochemical staining were used to examine liver pathologies,and the levels of IL-6,IL-1β,and TNF-αin the liver tissues were determined with ELISA.The mRNA expressions of HEM2M and the markers of M1 macrophages(TNF-α,iNOS,and IL-6)and M2 macrophages(Arg-1,YM-1,and IL-10)were detected using qRT-PCR,and the protein expressions of P-AKT,T-AKT,NLRC4,caspase-1 and GSDMD were assayed using immunoblotting.Caspase-1 activity in the liver tissues was determined with colorimetric measurement and immunofluorescence assay.Results Compared with HFD-fed WT mice,MYKI mice with HFD feeding showed milder liver function damage(P<0.01),alleviated hepatic steatosis,and reduced liver macrophage infiltration,glucose tolerance impairment and insulin resistance(P<0.01).The levels of IL-6,IL-1β,and TNF-αand mRNA expressions of M1 type macrophage markers were significantly decreased(P<0.01)and those of M2 type markers increased(P<0.01)in the liver tissues of HFD-fed MYKI mice,which also showed reduced NLRC4 inflammasome activity,caspase-1 activation,and GSDMD-N protein expression compared with their WT counterparts(P<0.05).Conclusion Overexpression of HEM2M reduces the production of hepatic inflammatory factors,improves insulin resistance and inhibits hepatic NLRC4 inflammasome activation,which leads to reduced hepatic pyroptosis and liver injury in NAFLD mice.

关 键 词：lncRNAHEM2M 巨噬细胞 非酒精性脂肪肝 细胞焦亡 

分 类 号：R575.5[医药卫生—消化系统]