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目的观察不同浓度叶酸对无机砷所致肝细胞毒性和氧化应激状态的影响。方法分别以无叶酸(0μmol/L)、正常叶酸(2.3μmol/L)、高叶酸(10μmol/L)培养液和20μmol/L亚砷酸钠(NaAsO2)对Chang肝细胞进行处理。染砷24h后,以3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞活力,以流式细胞术AnnexinV/PI双染法检测细胞凋亡情况,以2’,7’-二乙酰二氯荧光素(DCFH-DA)测定细胞内活性氧(ROS)水平,分别用DTNB法和硫代巴比妥酸比色法测定细胞内还原型谷胱甘肽(GSH)和丙二醛(MDA)含量。结果NaAsO2所致细胞活力降低和细胞内ROS、MDA相对水平的升高在无叶酸培养条件下显著加剧(P<0.05),在高叶酸条件下显著减轻(P<0.05);NaAsO2诱导的细胞内GSH相对含量升高在无叶酸条件下显著减弱(P<0.05),在高叶酸环境下显著增强(P<0.05);NaAsO2诱导的凋亡细胞在无叶酸条件下显著增多(P<0.05)。结论叶酸可改善无机砷所致氧化应激,减轻无机砷的肝细胞毒性。
Objective To observe the effects of different concentrations of folic acid on hepatotoxicity and oxidative stress induced by inorganic arsenic. Methods Chang hepatocytes were treated with folic acid (0μmol / L), normal folic acid (2.3μmol / L), folic acid (10μmol / L) and 20μmol / L sodium arsenite (NaAsO2) respectively. The cells were stained with 3- (4,5-dimethylthiazol-2) -2,5-diphenyltetrazolium bromide (MTT) for 24 h after arsenic exposure. Flow cytometry was performed using AnnexinV / Cell apoptosis was detected by staining method. The level of intracellular reactive oxygen species (ROS) was determined by 2 ’, 7’-diacetyl dichlorofluorescein (DCFH-DA). The cell viability was measured by DTNB method and thiobarbituric acid colorimetry The levels of intracellular reduced glutathione (GSH) and malondialdehyde (MDA) were determined. Results The decrease of cell viability induced by NaAsO2 and the relative increase of intracellular ROS and MDA were significantly increased (P <0.05) in the absence of folic acid and significantly decreased in the condition of high folic acid (P <0.05). NaAsO2 induced intracellular The increase of relative content of GSH was significantly attenuated in the absence of folic acid (P <0.05) and significantly increased in the folic acid (P <0.05). The apoptotic cells induced by NaAsO2 were significantly increased in the absence of folic acid (P <0.05). Conclusion Folic acid can improve the oxidative stress caused by inorganic arsenic and reduce the hepatotoxicity of inorganic arsenic.