miR-223/Fosl2/NGP调控非酒精性脂肪性肝炎小鼠肝纤维化作用  

The regulatory effect of miR-223/Fosl2/NGP on liver fibrosis in mice with non-alcoholic steatohepatitis

作　　者：王航宇 杨文义[1] 韩大正[1] 魏书堂[1] 武利萍[1] 徐梦阳 闫春晓[1] 谭莉霞 董勇 杨国威 华静 杜莹莹 WANG Hangyu;YANG Wenyi;HAN Dazheng;WEI Shutang;WU Liping;XU Mengyang;YAN Chunxiao;TAN Lixia;DONG Yong;YANG Guowei;HUA Jing;DU Yingying(Digestive Disease Combined Minimally Invasive Ward,the First Affiliated Hospital of Henan University,Kaifeng 475001,China)

机构地区：[1]河南大学第一附属医院消化病联合微创病区,河南开封475001

出　　处：《安徽医学》2022年第4期371-376,共6页Anhui Medical Journal

基　　金：河南省科技攻关计划项目(项目编号:LHGJ20190531,LHGJ20190532)。

摘　　要：目的探讨miR-223/FOS样抗原2(Fosl2)/中性粒细胞颗粒蛋白(NGP)通路调控非酒精性脂肪性肝炎(NASH)小鼠肝纤维化的作用。方法将60只小鼠随机分成正常组、蛋氨酸胆碱缺乏(MCD)组、MCD+miR-NC组(MCD模型鼠经尾静脉注射120 nmol/kg miR-NC 100μL)和MCD+miR-223 mimic组(MCD模型鼠经尾静脉注射120 nmol/kg miR-223 mimic 100μL),每组15只。正常组给予普通饲料,其余组给予MCD饲料。酶联免疫吸附法检测4组小鼠血清总胆固醇(TC)、三酰甘油(TG)、谷丙转氨酶(ALT)和谷草转氨酶(AST)水平;采用放射免疫分析法检测小鼠血清透明质酸(HA)、层粘连蛋白(LN)、Ⅳ型胶原(Ⅳ-C);实时荧光定量PCR检测miR-223和Fosl2 mRNA水平;观察小鼠肝脏病理变化;Western blot检测肝组织Fosl2、NGP蛋白水平。结果与MCD组和MCD+miR-NC组相比,MCD+miR-223 mimic组中miR-223表达水平升高,Fosl2表达水平下降(P<0.05)。与MCD组和MCD+miR-NC组相比,MCD+miR-223 mimic组肝细胞空泡化现象减少,肝细胞肥大程度降低。与MCD组和MCD+miR-NC组相比,MCD+miR-223 mimic组TC、TG、ALT和AST水平下降(P<0.05)。与MCD组和MCD+miR-NC组相比,MCD+miR-223 mimic组中脂质滴减少(P<0.05)。与MCD组和MCD+miR-NC组相比,MCD+miR-223 mimic组HA、LN和Ⅳ-C水平降低(P<0.05)。与MCD组和MCD+miR-NC组相比,MCD+miR-223 mimic组Fosl2蛋白水平下降,NGP蛋白水平上升(P<0.05)。结论miR-223可能通过调控Fosl2/NGP通路减轻NASH小鼠肝纤维化程度。Objective To explore the regulatory effect of miR-223/FOS-like antigen 2(Fosl2)/neutrophil granule protein(NGP)pathways on liver fibrosis in mice with non-alcoholic steatohepatitis(NASH).Methods A total of 60 mice were randomly divided into normal group,methionine choline deficiency(MCD)group,MCD+miR-NC group[MCD mice were injected with 100μL of miR-NC(120 nmol/kg)via tail vein]and MCD+miR-223 mimic group[MCD mice were injected with 100μL of miR-223 mimic(120 nmol/kg)via tail vein],with 15 cases in each group.The normal group was given common feed,while the other groups were given MCD feed.The levels of serum total cholesterol(TC),triglyceride(TG),alanine transaminase(ALT)and aspartate transaminase(AST)were detected by enzyme-linked immunosorbent assay.The levels of serum hyaluronic acid(HA),laminin(LN)and collagen typeⅣ(Ⅳ-C)were detected by radioimmunoassay.The levels of miR-223 and Fosl2 mRNA were detected by real-time fluorescence quantitative PCR.The pathological changes of liver tissues were observed.The levels of Fosl2 and NGP in liver tissues were detected by Western blot.Results Compared with MCD group and MCD+miR-NC group,the expression level of miR-223 increased,while the expression level of Fosl2 decreased in MCD+miR-223 mimic group(P<0.05).Compared with MCD group and MCD+miR-NC group,the hepatocyte vacuolation was reduced,and hepatocyte hypertrophy was relieved in MCD+miR-223 mimic group.Compared with MCD group and MCD+miR-NC group,the levels of TC,TG,ALT and AST decreased in MCD+miR-223 mimic group(P<0.05).Compared with MCD group and MCD+miR-NC group,lipid droplets were reduced in MCD+miR-223 mimic group(P<0.05).Compared with MCD group and MCD+miR-NC group,the levels of HA,LN andⅣ-C decreased in MCD+miR-223 mimic group(P<0.05).Compared with MCD group and MCD+miR-NC group,Fosl2 level decreased,while NGP level increased in MCD+miR-223 mimic group(P<0.05).Conclusion The miR-223 may relieve liver fibrosis in NASH mice by regulating Fosl2/NGP pathways.

关 键 词：脂肪性肝炎 非酒精性 肝纤维化 微小RNA223 FOS样抗原2 中性粒细胞颗粒蛋白 

分 类 号：R575.5[医药卫生—消化系统]