维生素K2降低体外星形胶质细胞的缺氧性损伤

Vitamin K2 attenuates hypoxia injury of astrocytes in vitro

目的:研究在缺氧环境下,维生素K2(vitamin K2,VK2)对星形胶质细胞的保护作用。方法 :利用新生SD大鼠大脑皮层进行星形胶质细胞培养,用5-二苯基四氮唑溴盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-Htetrazolium bromide,MTT]法检测VK2对缺氧星形胶质细胞活性的影响。用二氯二氢荧光素-乙酰乙酸酯(2',7'-dichlorofluorescin diacetate,DCF-DA)和二氢乙锭(dihydroethidium,DHE)染色检测VK2对缺氧条件下星形胶质细胞内活性氧(reactive oxygen species,ROS)的含量和超氧化物产生的作用。用实时聚合酶链式反应(real time polymerase chain reaction,RT-PCR)检测VK2处理的缺氧星形胶质细胞内生长停滞特异性基因6(growth arrest-specific gene 6,Gas6)和抗凝血因子蛋白质S(protein S)的m RNA表达量变化。结果:在0.5μmol/L和1μmol/L浓度下VK2显著提高缺氧条件下星形胶质细胞的活性。VK2使缺氧星形胶质细胞内ROS浓度降低及Gas 6和protein S的基因表达量升高。结论:在缺氧环境下,VK2可以提高星形胶质细胞的活性和降低ROS和超氧化物的产生,而Gas6和protein S可能参与此作用过程。

Objective:To explore the role of vitamin K2(VK2)inprotecting astrocyte growth in vitro under hypoxic conditions.Methods:The astrocytes from cerebral cortex of newborn SD rats were cultured in plates.The3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay was performed to assess the effect of VK2on cell viability in hypoxic conditions.Also2’,7’-dichlorofluorescin diacetate(DCF-DA)and dihydroethidium(DHE)stainings were used to detect intracellular radical oxygen species(ROS)production and superoxide in hypoxic astrocytes when treated with or without VK2.Real time polymerase chain reaction(RT-PCR)was used to detect mRNA expression of growth arrest-specific gene6(Gas6)and protein S in hypoxic astrocytes when treated with or without VK2.Results:The proliferation of astrocytes under hypoxic conditions were less reduced when treated with0.5μmol/L and1μmol/L VK2.The VK2also significantly reduced ROS concentration and increased the mRNA expression of Gas6and protein S in hypoxia astrocytes.Conclusion:VK2can promote the astrocytes viability under hypoxic conditions.The protection role may be through reducing the ROS as well as superoxide generation,Gas6and protein S are invdved in the effect.