miR-29a对MPP + 诱导PC12细胞凋亡影响及机制

EFFECT AND MECHANISM OF ACTION OF MICRORNA-29A ON 1-METHYL-4-PHENYLPYRIDINIUM-INDUCED APOPTOSIS OF PC12 CELLS

目的研究微小RNA-29a(miR-29a)对1-甲基-4-苯基吡啶离子(MPP^+)诱导的PC12细胞氧化应激和凋亡的影响以及作用机制。方法不同浓度MPP^+(0、100、300、500μmol/L)诱导PC12细胞24 h,荧光定量PCR(qPCR)检测miR-29a的表达量,噻唑蓝比色法(MTT)检测细胞活力。PC12细胞分别采用0和500μmol/L MPP^+干预并转染anti-miR-29a或对照质粒,采用流式细胞术检测细胞的凋亡率。观察细胞活性氧(ROS)水平。TargetScan软件预测、双荧光素酶报告基因验证miR-29a与重组人转化生长因子诱导因子同源框2(TGIF2)的靶向关系。结果随着MPP+浓度的增加miR-29a表达量随之增加(F=590.067,P<0.05),而细胞活力则随之降低(F=153.561,P<0.05)。下调miR-29a表达可抑制PC12细胞凋亡(F=301.044,P<0.05),降低ROS水平(F=254.120,P<0.05)。TGIF2是miR-29a下游靶基因。结论miR-29a可能通过调控TGIF2抑制MPP^+诱导的PC12细胞氧化应激和凋亡。

Objective To investigate the effect and mechanism of action of microRNA-29a(miR-29a)on 1-methyl-4-phenylpyridinium(MPP+)-induced oxidative stress and apoptosis of PC12 cells.Methods PC12 cells were induced by different concentrations of MPP^+(0,100,300,and 500μmol/L)for 24 hours,and then quantitative real-time PCR was used to measure the expression of miR-29a and MTT assay was used to measure cell viability.PC12 cells were treated with MPP+(0 and 500μmol/L)and transfected with anti-miR-29a or control plasmids,and flow cytometry was used to measure the apoptosis rate of the cells.The level of reactive oxygen species(ROS)in cells was observed.TargetScan software was used to predict and dual-luciferase reporter genes were used to verify the targeting relationship between miR-29a and recombinant human TGFB-induced factor homeobox 2(TGIF2).Results The expression level of miR-29a increased with the increase in the concentration of MPP^+(F=590.067,P<0.05),while cell viability decreased with the increase in the concentration of MPP^+(F=153.561,P<0.05).Downregulation of miR-29a expression inhibited the apoptosis of PC12 cells(F=301.044,P<0.05)and reduced the level of ROS(F=254.120,P<0.05).TGIF2 was a downstream target gene of miR-29a.Conclusion miR-29a may inhibit MPP^+-induced oxidative stress and apoptosis of PC12 cells by regulating TGIF2.