miR-204-5p靶向调控BCL2L2促进急性肝衰竭大鼠肝细胞凋亡的研究  

Study on miR-204-5p Promotes Hepatocyte Apoptosis in Acute Liver Failure Rats by Targeting BCL2L2

作　　者：王坤 朱传龙[2] 田健美[1] 孔小行[1] 成芳芳[1] 李嫣[1] WANG Kun;ZHU Chuanlong;TIAN Jianmei(Children′s Hospital of Soochow University,Jiangsu 215000,China)

机构地区：[1]苏州大学附属儿童医院感染性疾病科,215000 [2]南京医科大学第一附属医院感染病科,210029

出　　处：《医学研究杂志》2023年第7期136-141,共6页Journal of Medical Research

基　　金：江苏省苏州市科技发展计划项目(SZS2020310)。

摘　　要：目的探讨miR-204-5p在急性肝衰竭大鼠肝组织中的表达情况及其促进肝细胞凋亡的的作用及潜在机制。方法根据随机数字表法将32只SD大鼠分为4组,即对照组(0h)、模型组(24、48、72h),每组8只。模型组腹腔注射D-氨基半乳糖联合脂多糖诱导急性肝衰竭,对照组为腹腔内注射等量的0.9%氯化钠溶液。分别取对照组及模型组造模完成后24、48、72h大鼠静脉血及肝组织,ELISA检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸基转移酶(aspartate aminotransferase,AST)、总胆红素(total bilirubin,TBIL)的水平,通过HE、TUNEL染色法行肝组织病理学检查和肝细胞凋亡观察。生物信息学工具预测miR-204-5p的靶基因,采用双荧光素酶报告基因实验验证两者的靶向关系。RT-PCR和Western blot法检测肝组织中miR-204-5p、BCL2L2的表达情况。结果与对照组比较,模型组随着造模时间的延长,ALT、AST、TBIL值逐渐升高,肝组织坏死程度、炎性细胞浸润逐渐加重,肝细胞凋亡指数逐渐升高。RT-PCR结果显示,随着造模时间的延长,miR-204-5p基因的相对表达水平逐渐升高,而BCL2L2基因的相对表达水平逐渐降低;Western blot法检测结果亦显示,BCL2L2蛋白表达水平逐渐降低(P均<0.05)。miR-204-5p和BCL2L2之间存在靶向关系,miR-204-5p能够靶向调控BCL2L2表达。结论miR-204-5p可能通过靶向调控BCL2L2促进肝细胞凋亡,进而促进急性肝衰竭疾病的发生、发展。Objective To explore the expression of miR-204-5p in liver tissue of rats with acute liver failure,as well as its role in promoting hepatocyte apoptosis and potential mechanisms.Methods Thirty-two SD rats were randomly divided into four groups[control group(0h),model groups(24h,48h and 72h)],each group had eight rats.Model groups were intraperitoneally injected with D-galactosamine(D-GaIN)and lipopolysaccharide(LPS),while rats in the control group were injected with equal amount of saline.The venous blood and liver tissue of rats were obtained in the control group and the model groups which were taken in 24hours,48hours and 72hours after the completion of the model.The levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL)were tested by Enzyme-linked immunosorbent assay(ELISA).The pathological change of liver tissue was examined by HE staining and the apoptosis was observed by TUNEL.Bioinformatics tools were used to predict the target gene of miR-204-5p,and the dual luciferase reporter gene experiment was used to verify the targeting relationship.RT-PCR and Western blot were used to detect the levels of miR-204-5p,BCL2L2 in the liver tissues.Results Compared with the control group,with the prolongation of modeling time,the level of ALT,AST and TBIL in the model groups gradually increased.The degree of liver tissue necrosis and inflammatory cell infiltration gradually aggravated,and the apoptotic index of hepatocyte gradually rose.The results of RT-PCR showed that with the prolongation of modeling time,the relative expression level of miR-204-5p in liver tissue was upregulated,while BCL2L2 was downregulated.Western blot showed that the expression level of BCL2L2 protein decreased gradually(all P<0.05).There is a targeting relationship between miR-204-5p and BCL2L2,indicating that miR-204-5p can target the expression of BCL2L2.Conclusion miR-204-5p promotes hepatocyte apoptosis by targeting BCL2L2,which promoting the occurrence and development of acute liver failure.

关 键 词：急性肝衰竭 微小RNA-204-5p BCL2L2 凋亡 

分 类 号：R575.3[医药卫生—消化系统]