过氧化氢干预下Rab30在PC12细胞中的表达及促凋亡通路

Expression of Rab30 in PC12 cells and related pro-apoptotic pathways under hydrogen peroxide intervention

目的探讨经过氧化氢(H_2O_2)损伤干预下Rab30在PC12细胞中的表达及可能的促凋亡通路。方法将体外常规培养的PC12细胞分为正常组和H_2O_2组。采用MTT检测细胞存活率;Annexin V-FITC/PI双染法检测细胞凋亡率;Western-blot检测Rab30、JNK3、GM130、Caspase-3的蛋白表达情况。结果 H_2O_2损伤干预可导致PC12细胞的存活率显著降低(P<0.05),早期凋亡率和总凋亡率均显著增高(均P<0.05);JNK3、Caspase-3蛋白表达水平在PC12细胞中显著增加(P<0.05),而Rab30、GM130蛋白表达显著下降(P<0.05)。结论 H_2O_2在PC12细胞中的促凋亡通路可能是H_2O_2激活JNK3的表达而靶向下调Rab30,致使高尔基体结构破碎,并激活Caspase-3的活性,从而诱导PC12细胞的凋亡。

Objective To investigate the expression of Rab30 in PC12 cells and possible pro-apoptotic pathways under hydrogen perox- ide （H2O2 ） intervention. Methods PC12 cells cultured in vitro were divided into normal group and H2 02 group. MTT assay was used to measure cell viability, Annexin V-FITC/PI double staining was used to measure apoptosis rate, and Western blot was used to meas- ure the protein expression of Rab30, JNK3, GM130, and caspase-3. Results H202 intervention induced a significant reduction in the survival rate of PC12 cells （P 〈 0.05 ） and significant increases in early apoptosis rate and overall apoptosis rate （P 〈 0.05 ） ; the PC 12 cells had significant increases in the protein expression of JNK3 and easpase-3 （P 〈0.05 ） and significant reductions in the protein expression of Rab30 and GM130 （P 〈 0.05 ）. Conclusions The pro-apoptotic pathway of H2O2 in PC12 cells may be activation of JNK3 expression, targeted down-regulation of Rab30, destruction of Golgi structure, and activation of easpase-3, which induces the ap- optosis of PC12 cells.