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目的:探讨miR-34a在幼鼠海马神经元细胞增殖凋亡中的作用。方法:分离幼鼠海马神经元细胞,转染miR-34a抑制剂(miR-34a inhibitor)、抑制剂对照(inhibitor control)、miR-34a模拟物(miR-34a mimics)、模拟物对照(mimics control),RT-PCR检测细胞中miR-34a表达水平。MTT检测转染后细胞增殖情况。流式细胞仪检测细胞凋亡情况。Western blot检测细胞中Cleaved-caspase-3、Bcl-2、Bax的表达水平。结果:转染miR-34a inhibitor可以抑制miR-34a的表达,miR-34a mimics可以促进miR-34a的表达。miR-34a mimics对细胞增殖抑制率明显高于mimics control组(P<0.05),miR-34a inhibitor组抑制率明显低于inhibitor control组(P<0.05)。miR-34a inhibitor组神经元细胞凋亡率明显低于inhibitor control组(P<0.05),miR-34a mimics组神经元细胞凋亡率明显高于mimics control组(P<0.01),inhibitor control组和mimics control组神经元细胞凋亡率差异不显著(P>0.05)。miR-34a inhibitor组Cleaved-caspase-3、Bax蛋白表达量低于inhibitor control组,差异显著(P<0.05);miR-34a inhibitor组Bcl-2蛋白表达量高于inhibitor control组,差异显著(P<0.05);miR-34a mimics组Cleaved-caspase-3、Bax蛋白表达量高于mimics control,差异显著(P<0.05);miR-34a mimics组Bcl-2蛋白表达量低于mimics control,差异显著(P<0.05)。结论:miR-34a抑制海马神经元细胞增殖,促进细胞凋亡,其作用机制可能与调控Cleaved-caspase-3、Bcl-2、Bax表达有关。
AIM: To investigate the role of miR-34a in the proliferation and apoptosis of neurons in the hippocampus of infant rats. Methods: The hippocampal neurons were isolated and transfected with miR-34a inhibitor, inhibitor control, miR-34a mimics, mimics control ). The expression of miR-34a in cells was detected by RT-PCR. MTT detection of cell proliferation after transfection. Flow cytometry detection of apoptosis. Western blot was used to detect the expression of Cleaved-caspase-3, Bcl-2 and Bax. Results: The miR-34a inhibitor transfected miR-34a can inhibit the expression of miR-34a, miR-34a mimics can promote the expression of miR-34a. The inhibitory rate of miR-34a mimics on cell proliferation was significantly higher than that of mimics control group (P <0.05), and the inhibition rate of miR-34a inhibitor group was significantly lower than that of inhibitor control group (P <0.05). The apoptosis rate of neurons in miR-34a inhibitor group was significantly lower than that of inhibitor control group (P <0.05), the apoptosis rate of neuron in miR-34a mimics group was significantly higher than that in mimics control group (P <0.01) There was no significant difference in neuron apoptosis rate between mimics control group and control group (P> 0.05). The protein expression of Cleaved-caspase-3 and Bax in miR-34a inhibitor group was significantly lower than that in inhibitor control group (P <0.05). The expression of Bcl-2 protein in miR-34a inhibitor group was significantly higher than that in inhibitor control group <0.05). The expression of Cleaved-caspase-3 and Bax in miR-34a mimics group was higher than that of mimics control (P <0.05), while the expression of Bcl-2 protein in miR-34a mimics group was lower than that of mimics control (P <0.05). Conclusion: miR-34a inhibits the proliferation of hippocampal neurons and promotes apoptosis. The possible mechanism may be related to the regulation of the expression of Cleaved-caspase-3, Bcl-2 and Bax.