叶酸对亚砷酸钠致Chang肝细胞毒性和氧化应激的影响

Effects of Folate on Sodium Arsenite-Induced Toxicity and Oxidative Stress in Chang Hepatocytes

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 folatc levels on inorganic arsenic-induced cytotoxicity and oxidative stress status. Methods Chang human hepatocytes were cultured with 20 μmol/L sodium arscnite （NaAsO2） in medium containing 0 μmol/L, 2.3μmol/L and 10μ mol/L of folate, respectively. After 24 h of treatment with NaAsO2, cell viability was analyzed with 3- （4,5-dimethylthiazol-2-yl）-2,5-diphenyhetrazolium bromide （MTT） method, apoptosis was assessed by Annexin V/PI double staining, levels of ROS were detected by 2＇ ,7＇ -dichlorodihydrofluorescein diacetate （DCFH-DA） probe, and intracellular reduced glutathione （GSH） and malondialdehyde （MDA） content were determined by DTNB method and formation of thiobarbituric acid reactive substances, respectively. Results The decrease of cell viability and the increase of intracellular ROS and MDA induced by NaAsO2 were significantly exacerbated by folate deficiency but attenuated by folate supplement （P〈0.05）. Moreover, the NaAsO2-induced elevation of intracellular GSH was attenuated by folate deficiency but aggravated by folate supplement （P〈O.05）. Apoptosis cells induced by NaAsO2 were significantly increased by folate deficiency （P〈0.05）. Conclusion Folate can attenuate the oxidative stress and cytotoxicity induced by inorganic arsenic in hepatocytes.