SOX6基因对H2O2诱导的星形胶质细胞损伤的机制
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青海大学附属医院神经外科,西宁 810000

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R-33

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Mechanism of action of SOX6 gene on astrocyte injury induced by H2O2
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Department of Neurosurgery, Affiliated Hospital of Qinghai University, Xining 810000, China

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    摘要:

    目的 探讨干扰SOX6基因表达H2O2诱导的星形胶质细胞(AS)凋亡、线粒体膜电位及PI3K/AKT信号通路影响。 方法 从乳大鼠大脑皮质分离培养AS细胞,将细胞分为阴性对照组(NC组):A转染无干扰作用siRNA;H2O2组:20μmol / L的H2O2处理AS细胞24 h;H2O2+NC组:无干扰作用的siRNA转染AS细胞24 h,20mol / L的H2O2处理细胞24 h;H2O2+si-SOX6组:SOX6特异性siRNA转染AS细胞24 h后,20 μmol / L的H2O2处理细胞 24 h。通过LDH试剂盒、流式细胞术及JC-1探针分别检测细胞毒性、凋亡率及线粒体膜电位; Western blotting 检测SOX6、Cyt.C、caspase3、caspase9、Bcl-2、Bax和 p-AKT蛋白表达。 结果 H2O2 可明显上调AS细胞SOX6、Cyt.C、caspase3、caspase9和Bax表达,下调Bcl-2和 p-AKT 表达,提高细胞毒性,诱导细胞凋亡,而干扰SOX6 基因表达可明显减弱H2O2对细胞SOX6、Cyt.C、caspase3、caspase9、Bcl-2、Bax和p-AKT 表达及细胞毒性和凋亡影响。 结论 干扰 SOX6 基因表达可通过抗凋亡途径保护H2O2诱导的AS损伤,此保护作用与线粒体通路及PI3K/ AKT 信号通化激活有关。

    Abstract:

    Objective To investigate the effects of SOX6 gene interference on apoptosis, mitochondrial membrane potential, and the PI3K/ AKT signaling pathway of astrocytes (AS) induced by H2O2 . Methods AS cells were isolated from the cerebral cortex of neonatal rats and divided into the following groups: negative control group (NC group): AS transfected with siRNA without interference; H2O2 group: AS cells treated with 20 mol/LH2O2 for 24 h; H2O2 +NC group: non-interfering siRNA transfected into AS cells for 24 h, then treatment of cells with 20μmol /L H2O2 for 24 h;and H2O2+si-SOX6 group: SOX6-specific siRNA transfected into AS cells for 24 h, followed by treatment of the cells with 20 μmol / L H2O2 for 24 h. The cytotoxicity, apoptosis rate, and mitochondrial membrane potential were determined by LDH kit, flow cytometry, and JC-1 probe. The expression of SOX6, Cyt. C, caspase 3, caspase9, Bcl-2, Bax, and p-AKT protein was determined by Western blotting. Results H2O2 significantly upregulated the expression of SOX6, Cyt. C, caspase3, caspase9, and Bax; downregulated the expression of Bcl-2 and p-AKT; increased cytotoxicity; and induced apoptosis. Interfering with SOX6 gene expression significantly reduced the effects of H2O2 on SOX6, Cyt.C, caspase3, caspase9, Bcl-2, Bax, and p-AKT expression, cytotoxicity, and apoptosis. Conclusions Interference with SOX6 gene expression can protect against AS injury induced by H2O2 through the anti-apoptotic pathway, which is related to the mitochondrial pathway and PI3K/ AKT signaling activation.

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巨虎,肖宗宇,张广华,许常林.SOX6基因对H2O2诱导的星形胶质细胞损伤的机制[J].中国比较医学杂志,2020,30(1):88~93.

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  • 收稿日期:2019-06-10
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  • 在线发布日期: 2020-08-19
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