上调miR-19b表达对H2O2诱导SH-SY5Y细胞损伤影响及机制  

EFFECT OF MIR-19B UPREGULATION ON H2O2-INDUCED SH-SY5Y CELL INJURY AND RELATED MECHANISM

作　　者：代晓晓 鞠波[1] DAI Xiaoxiao;JU Bo(Department of Neurology,970 Hospital of PLA Joint Logistics Support Force,Yantai 264010,China)

机构地区：[1]中国人民解放军联勤保障部队第970医院神经内科,山东烟台264010

出　　处：《青岛大学学报（医学版）》2021年第1期82-86,共5页Journal of Qingdao University(Medical Sciences)

基　　金：山东省自然科学基金资助项目(ZR2017HL098)。

摘　　要：目的探讨上调miR-19b表达对H2O2诱导的SH-SY5Y细胞损伤的影响及机制。方法体外培养SH-SY5Y细胞,将其分为对照组、H2O2组、H2O2+miR-NC组和H2O2+miR-19b组,采用实时荧光定量PCR(RT-PCR)检测各组细胞中miR-19b表达,四甲基噻唑蓝(MTT)法和流式细胞仪分别检测细胞存活率和凋亡率,相关试剂盒检测细胞上清液中乳酸脱氢酶(LDH)、细胞内超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽还原酶(GSH-Px)水平,蛋白印迹(Western blot)检测各组细胞中磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)和磷酸化AKT(p-AKT)等蛋白表达。结果与对照组相比,H2O2组细胞中miR-19b表达、细胞存活率和细胞内SOD、GSH-Px活力及PI3K、p-AKT蛋白表达水平均降低(P均<0.05),而细胞凋亡率、LDH活力和MDA含量均升高(P均<0.05)。与H2O2组相比较,H2O2+miR-19b组miR-19b表达、细胞存活率和细胞内SOD、GSH-Px活力以及PI3K、p-AKT蛋白表达水平均升高(P均<0.05),而细胞凋亡率、LDH活力和MDA含量均降低(P均<0.05)。结论上调miR-19b表达可提高H2O2诱导的SH-SY5Y神经细胞存活率,减轻细胞凋亡和氧化应激损伤,其作用机制可能与激活PI3K/AKT信号通路有关。Objective To investigate the effect of miR-19b upregulation on H2O2-induced SH-SY5Y cell injury and rela-ted mechanism.Methods SH-SY5Y cells were cultured in vitro and were then divided into control group,H2O2 group,H2O2+miR-NC group,and H2O2+miR-19b group.RT-PCR was used to measure the expression of miR-19b in each group;MTT assay and flow cytometry were used to measure cell viability and apoptosis rate,respectively;related kits were used to measure the level of lactate dehydrogenase(LDH)in cell supernatant and the levels of superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-Px)in cells;Western blot was used to measure the protein expression of phosphoinositide 3-kinase(PI3K),protein kinase B(AKT),and phosphorylated AKT(p-AKT).Results Compared with the control group,the H2O2 group had significant reductions in the expression of miR-19b in cells,cell viability,the activities of SOD and GSH-Px in cells,and the protein expression levels of PI3K and p-AKT(all P<0.05),as well as significant increases in cell apoptosis rate,LDH activity,and MDA content(all P<0.05).Compared with the H2O2 group,the H2O2+miR-19b group had significant increases in the expression of miR-19b,cell viability,the activities of SOD and GSH-Px,and the protein expression levels of PI3K and p-AKT(all P<0.05),as well as significant reductions in cell apoptosis rate,LDH activity,and MDA content(all P<0.05).Conclusion Upregulation of miR-19b can increase the viability of H2O2-induced SH-SY5Y cells and reduce cell apoptosis and oxidative stress injury,possibly by activating the PI3K/AKT signaling pathway.

关 键 词：miR-19b 神经细胞 细胞凋亡 氧化应激 PI3K/AKT信号通路 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学] R342.2[医药卫生—基础医学]