miRNA-122-5p对细胞色素P4503A4调控替格瑞洛代谢细胞学研究  

miRNA-122-5p regulates ticagrelor metabolism on cytochrome P4503A4 cytological study

作　　者：才艺 赵晓杰 张效林 刘丹 田孝祥 闫承慧 CAI Yi;ZHAO Xiao-jie;ZHANG Xiao-lin;LIU Dan;TIAN Xiao-xiang;YAN Cheng-hui(Department of Cardiovascular,General Hospital of Northern Theater Command,Shenyang 110016,China)

机构地区：[1]北部战区总医院心血管内科,辽宁沈阳110016

出　　处：《临床军医杂志》2021年第10期1122-1126,共5页Clinical Journal of Medical Officers

基　　金：辽宁省自然科学基金(2019020024-JH2/103);辽宁省自然科学基金(2019-ZD-1060);辽宁省自然科学基金(2019-ZD-1038)。

摘　　要：目的探讨miRNA-122-5p是否通过特异性调控人正常肝细胞(LO2)中细胞色素P4503A4(CYP3A4)的表达,从而影响替格瑞洛的代谢。方法采用LO2进行4组细胞转染实验:miR-122-5p模拟物组、miR-122-5p模拟物阴性对照组、miR-122-5p抑制剂组、miR-122-5p抑制剂阴性对照组。通过蛋白质印迹和实时聚合酶链反应鉴定CYP3A4蛋白和转录的表达水平。在miR-122-5p模拟物和miR-122-5p抑制剂转染后,采用5μmol/L替格瑞洛刺激LO224 h。通过高效液相质谱联用检测细胞培养基中替格瑞洛及其活性代谢物AR-C124910XX的浓度。结果在蛋白表达水平方面,与miR-122-5p模拟物阴性对照组相比,miR-122-5p模拟物组CYP3A4的表达降低(P<0.05)。与miR-122-5p抑制剂阴性对照组相比,miR-122-5p抑制剂组CYP3A4的表达增加(P<0.05)。在转录水平上,与miR-122-5p模拟物阴性对照组相比,miR-122-5p模拟物转染组CYP3A4的表达降低(P<0.05)。与miR-122-5p抑制剂阴性对照组相比,miR-122-5p抑制剂组CYP3A4的表达增加(P<0.05)。与miR-122-5p模拟物阴性对照组相比,miR-122-5p模拟物组细胞培养基中替格瑞洛的浓度增高,其活性代谢物AR-C124910XX的浓度降低(P<0.05)。与miR-122-5p抑制剂阴性对照组相比,miR-122-5p抑制剂组细胞培养基中替格瑞洛的浓度降低,其活性代谢物AR-C124910XX的浓度增加(P<0.05)。结论miR-122-5p参与CYP3A4表达的调控,并影响替格瑞洛的代谢。Objective To investigate whether miR-122-5 P specifically regulates the expression of cytochrome P4503 A4(CYP3 A4)in human normal liver cells(LO2),thereby affecting the metabolism of ticagrelor.Methods Four groups of cells were transfected with LO2:miR-122-5 p mimic group,miR-122-5 p mimic negative control group,miR-122-5 p inhibitor group,and miR-122-5 p inhibitor negative control group.The expression levels of CYP3 A4 protein and transcription were determined by western blotting and real-time polymerase chain reaction.After transfection with miR-122-5 p mimic and miR-122-5 p inhibitors,LO2 cells were stimulated with 5μmol/L ticagrelor for 24 h.The concentrations of ticagrelor and its active metabolite AR-C124910 XX in cell culture medium were determined by high performance liquid chromatography(HPLC)-mass spectrometry.Results In terms of protein expression level,CYP3 A4 expression in miR-122-5 p mimic group was significantly decreased compared with miR-122-5 p mimic negative control group(P<0.05).Compared with miR-122-5 p inhibitor negative control group,the expression of CYP3 A4 in miR-122-5 p inhibitor group was significantly increased(P<0.05).At the transcriptional level,CYP3 A4 expression was significantly decreased in miR-122-5 p mimic transfected group compared with miR-122-5 p mimic negative control group(P<0.05).Compared with miR-122-5 p inhibitor negative control group,the expression of CYP3 A4 in miR-122-5 p inhibitor group was significantly increased(P<0.05).Compared with miR-122-5 p mimic negative control group,the concentration of ticagrelor in cell culture medium in miR-122-5 p mimic group was significantly increased,and the concentration of its active metabolite AR-C124910 XX was significantly decreased(P<0.05).Compared with miR-122-5 p inhibitor negative control group,the concentration of ticagrelor in cell culture medium in miR-122-5 p inhibitor group was significantly decreased,and the concentration of its active metabolite AR-C124910 XX was significantly increased(P<0.05).Conclusion miR-122-

关 键 词：miR-122-5p 细胞色素P4503A4 替格瑞洛 高效液相质谱联用 

分 类 号：R541.4[医药卫生—心血管疾病]