摘要
目的探讨水翁花对过氧化氢(H2O2)诱导神经细胞PC12凋亡的影响及其作用机制。方法构建PC12氧化应激损伤模型,不同浓度的水翁花干预H2O2诱导的PC12细胞,应用流式细胞仪检测细胞凋亡率;检测细胞SOD水平。qRT PCR法检测水翁花对miR 422a表达的影响;过表达miR 422a或抑制miR 422a表达对H2O2诱导的PC12细胞凋亡及SOD活性的影响。Western blot法测定细胞中Bcl 2、Bax的表达变化。结果H2O2诱导的PC12细胞凋亡率升高(P<0.05),SOD活性降低(P<0.05),抑制miR 422a表达(P<0.05),而水翁花处理后,PC12细胞凋亡率降低(P<0.05),SOD活性增强(P<0.05),促进miR 422a表达(P<0.05),Bcl 2表达上调(P<0.05),Bax表达下调(P<0.05);过表达miR 422a可抑制H2O2诱导的PC12细胞凋亡(P<0.05),增强SOD活性(P<0.05);抑制miR 422a表达可逆转水翁花对H2O2诱导的PC12细胞凋亡、SOD活性的影响。结论水翁花可通过上调miR 422a表达对H2O2诱导的PC12细胞损伤发挥保护作用,其可能作用机制与抗细胞凋亡及抗氧化作用相关。
Objective To investigate the effect of cleistocalyx operculatus flower buds on the apoptosis of PC12 induced by hydrogen peroxide(H2O2)and its mechanism.Methods PC12 oxidative stress injury model was established.Different concentrations of cleistocalyx operculatus flower buds intervened in H2O2 induced PC12 cells,and apoptosis rate was detected by flow cytometry.Cell SOD levels were measured according to the kit.The effect of operculatus flower on the expression of miR 422a was detected by qRT PCR.Overexpression of miR 422a or inhibition of miR 422a expression on H2O2 induced apoptosis and SOD activity in PC12 cells were measured.The expression of Bcl 2 and Bax in cells was determined by Western blot.Results H2O2 induced apoptosis of PC12 cells increased(P<0.05),SOD activity decreased(P<0.05),and miR 422a expression was inhibited(P<0.05).After treatment with water flower,the apoptosis rate of PC12 cells decreased,the activity of SOD increased(P<0.05),the expression of miR 422a and Bcl 2 was up regulated(P<0.05).The expression of Bax was down regulated(P<0.05).Overexpression of miR 422a inhibited H2O2 induced apoptosis of PC12 cells and enhanced SOD activity.Inhibition of miR 422a expression reversed the effect of water flower on H2O2 induced apoptosis and SOD activity in PC12 cells.Conclusion Cleistocalyx operculatus flower buds can protect H2O2 induced PC12 cell injury by up regulating miR 422a expression,and the possible mechanism is related to anti apoptosis and anti oxidation.
作者
于广周
谭文刚
张昊
YU Guangzhou;TAN Wengang;ZHANG Hao(Henan Province Zhumadian Central Hospital Sankei Third Branch,Zhumadian,Henan,China,463000)
出处
《分子诊断与治疗杂志》
2019年第6期517-521,544,共6页
Journal of Molecular Diagnostics and Therapy
关键词
水翁花
过氧化氢
MIR
422a
PC12细胞
凋亡
Cleistocalyx operculatus flower buds
hydrogen peroxide
miR 422a
PC12 cells
apoptosis
