过表达miR-122对异氟烷致PC12细胞损伤的保护作用  

Overexpression of miR-122 Protects PC12 Cellsfrom Damages Caused by Isoflurane

作　　者：陈霞[1] 陆丽[1] 耿明倩 李志刚 吴皎卿[2] CHEN Xia;LU Li;GENG Mingqian;LI Zhigang;WU Jiaoqing(Department of Anesthesiology, BenQ Hospital, Nanjing Medical University, Nanjing 210000, China;Department of Anesthesiology, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing 211500, China)

机构地区：[1]南京医科大学附属明基医院麻醉科,江苏南京210000 [2]南京医科大学第四附属医院麻醉科,江苏南京211500

出　　处：《标记免疫分析与临床》2021年第3期505-509,共5页Labeled Immunoassays and Clinical Medicine

摘　　要：目的探讨miR-122在异氟烷导致的PC12细胞损伤中的作用。方法将PC12细胞分为对照组、异氟烷组、阴性对照组(NC组)和miR-122过表达组,除对照组外,其余组细胞均用2%异氟烷+21%O_(2)+5%CO_(2)处理6h,miR-122过表达组和NC组在用异氟烷处理前通过脂质体2000将miR-122 mimic和mimic-NC转染细胞。通过RT-qPCR检测各组细胞中miR-122表达水平;MTT检测各组细胞处理0、24、48、72h后的细胞活力;ELISA试剂盒检测各组细胞MDA、SOD、GSH-Px水平;通过流式细胞仪检测各组细胞凋亡情况;Western-blot检测各组细胞Bax、Bcl-2、cleaved-Caspase-3蛋白表达。结果与对照组相比,异氟烷组细胞中miR-122相对表达量、24、48、72h细胞吸光度值、Bcl-2蛋白表达、SOD和GSH-Px水平下降(均P<0.05),MDA水平、细胞凋亡率、Bax和cleaved-Caspase-3蛋白表达升高(均P<0.05)。与异氟烷组相比,miR-122过表达组miR-122相对表达量、24、48、72h细胞吸光度值、Bcl-2蛋白表达、SOD和GSH-Px水平显著升高(P<0.05),MDA水平、细胞凋亡率、Bax和cleaved-Caspase-3蛋白表达下降(P<0.05)。结论过表达miR-122能够改善异氟烷诱导的PC12细胞活力降低和氧化应激水平,并抑制细胞凋亡。Objective To investigate the role of miR-122 in PC12 cell trauma caused by isoflurane.Methods PC12 cells were divided into the control group,isoflurane group,negative control group(NC group)and miR-122 over expression group.Except for the control group,all other groups were treated with 2%isoflurane+21%O_(2)+5%CO_(2) for 6 hours.miR-122 mimics and mimics NC were transfected into PC12 cells by Liposomes 2000 in the miR-122 overexpression group and NC group before treatment with isoflurane.The expression of miR-122 in each group of cells was detected by RT-qPCR.MTT was applied to detect the cell viability in each group of cells after 0 h,24 h,48 h,and 72 h.The contents of MDA,SOD and GSH-PX were detected by ELISA kit.Cell apoptosis was detected by flow cytometry.The expressions of Bax,Bcl-2 and cleaved-Caspase-3 were detected by western blot.Results Compared with the control group,the relative expression of miR-122,cell absorbance at 24 h,48 h and 72 h,Bcl-2 protein expression,SOD and GSH-PX contents in the isoflurane group were all decreased(all P<0.05),while MDA content,apoptosis rate,Bax and cleaved-Caspase-3 protein expressions were all increased(P<0.05).Compared with the isoflurane group,the relative expression of miR-122,cell absorbance at 24 h,48 h,and 72 h,Bcl-2 protein expression,SOD and GSH-PX contents in the miR-122 over expression group were all significantly increased(P<0.05),while MDA content,apoptosis rate,Bax and cleaved-Caspase-3 protein expressions were all decreased(P<0.05).Conclusion Overexpression of miR-122 can relieve isoflurane-induced inhibition of PC12 cell proliferation and oxidative stress,and inhibit cell apoptosis.

关 键 词：MIR-122 异氟烷 PC12细胞 细胞损伤 氧化应激 

分 类 号：Q786[生物学—分子生物学]