miR-125b对人心肌细胞HCM的保护作用及机制  

Protective effect and mechanism of miR-125b on human cardiomyocytes HCM

作　　者：解小霞[1] 孙晓静[1] 苏文炀 杨柳[1] 李力[1] 王薇[1] 李婷[1] 田永强 XIE Xiaoxia;SUN Xiaojing;SU Wenyang;YANG Liu;LI Li;WANG Wei;LI Ting;TIAN Yongqiang(Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430014,China)

机构地区：[1]武汉市中医医院,武汉430014 [2]湖北省中医院

出　　处：《山东医药》2020年第5期1-5,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81803676)

摘　　要：目的探讨微小RNA(miR)125b对人心肌细胞HCM的保护作用及其可能的作用机制。方法选取急性心肌梗死(AMI)患者68例为AMI组、体检健康对照者50例作为正常对照组,采集静脉血检测血清miR-125b表达。取对数生长期人心肌HCM细胞,分别采用瞬时转染法转染miR-125b mimics(浓度分别为200、100、50 nmol/L)及NC mimics(浓度分别为200、100、50 nmol/L),培养24 h及48 h,采用CCK-8法测定细胞相对增殖率。分别转染miR-125b mimics终浓度为50、100 nmol/L,转染NC mimics终浓度为100 nmol/L及空白组,采用流式细胞术测定细胞早期凋亡百分比。采用Western blotting法检测细胞免疫、炎症相关蛋白炎性小体(NLRP3)、白细胞介素1β(IL-1β)、趋化因子2(CCL2)、趋化因子C-X3-C-配体1(CX3CL1)、补体应答基因-32(RGC32)表达。结果AMI组血清miR-125b表达水平低于正常对照组(P<0.01)。24、48 h时,转染各浓度miR-125b后HCM细胞相对增殖率均高于转染NC mimics后(P均<0.05)。转染50、100 nmol/L miR-125b mimics后HCM细胞的早期凋亡百分比均低于转染NC mimics后(P<0.05或0.01)。转染miR-125b mimics 200 nmol/L后HCM细胞内NLRP3、IL-1β、CCL2、CX3CL1、RGC32的表达量均较NC mimics组降低(P均<0.05)。结论miR-125b可促进HCM细胞的增殖、抑制HCM细胞凋亡,其机制可能为下调免疫、炎症调节相关因子NLRP3、IL-1β、CCL2、CX3CL1、RGC32的表达,抑制NLRP3/IL-1β通路激活。Objective To investigate the protective effect of miR-125b on human cardiomyocytes HCM and to reveal its possible mechanism.Methods The relative expression of miR-125b in serum of 55 cases of healthy controls(control group)and 68 cases of acute myocardial infarction(AMI)patients(AMI group)was detected by qPCR.The miRNA-125b mimimics were transfected into HCM cells.Human cardiomyocytes HCM in the logarithmic phase were transfected with miR-125b mimics(200,100,and 50 nmol/L,respectively)and NC mimics(200,100,and 50 nmol/L,respectively)by transient transfection,and then were culture for 24 and 48 h.Flow cytometry was used to detect the apoptosis of HCM.Targetscan bioinformatics prediction software was used to predict the expression of miRNA-125b.The interaction between miRNA-125b and NLRP3 was detected by double luciferase reporter gene system.The effects of miRNA-125b on the protein expression of inflammatory corpuscle(NLRP3),interleukin-1β(IL-1β),chemokine 2(CCL2),chemokine C-X3-C-ligand 1(CX3CL1),complement response gene-32(RGC32)in HCM cells were detected by Western blotting.Results The expression of serum mRNA-125b in AMI group was significantly lower than that in the healthy control group(P<0.01).At 24 and 48 h,the relative proliferation rates of HCM cells after transfection with miR-125b at different concentrations were higher than those after transfection with NC mimics(all P<0.05).The percentage of early apoptosis in HCM cells after transfection with 50 and 100 nmol/L miR-125b mimics was lower than that after transfection with NC mimics(P<0.05 or P<0.01).After 200 nmol/L miR-125b mimics transfection,the expression levels of NLRP3,IL-1β,CCL2,CX3CL1,and RGC32 in HCM cells were all lower than those after NC mimics transfection(all P<0.05).Conclusion The miRNA-125b could promote the proliferation,inhibit the apoptosis of HCM cells by down-regulating the expression of NLRP3,IL-1β,CCL2,CX3CL1 and RGC32,and inhibiting the activation of NLRP3/IL-1βpathway.

关 键 词：心肌梗死 人心肌细胞 miR-125b NLRP3 CCL2 CX3CL1 RGC32 

分 类 号：R542.2[医药卫生—心血管疾病]