竹荪多糖对亚砷酸钠致人肝细胞毒性的拮抗作用  

Antagonism of dictyophora polysaccharide to sodium arsenite-induced human hepatocytes toxicity

作　　者：杨红艳[1] 罗鹏[1,2] 曾丹[1] 沈祥春[3] 胡婷[1] 肖智文 谢琅[1] 黄元庆[1] 

机构地区：[1]贵州医科大学公共卫生学院,贵州贵阳550000 [2]省部共建教育部重点实验室环境污染与疾病监控实验室 [3]贵州医科大学药学院

出　　处：《环境与健康杂志》2016年第6期493-497,共5页Journal of Environment and Health

基　　金：国家自然科学基金(81160336);贵州省科技厅农业攻关计划项目(黔科合NY字[2011]3055号);贵州省科技厅(黔科合J字[2010]2188)贵州省教育厅基金项目(黔省专合字[2010]39号)

摘　　要：目的探讨竹荪多糖对亚砷酸钠致人胎肝（L-02）细胞毒性的拮抗作用。方法将处于对数生长期的L-02细胞分别暴露于含不同浓度[0（对照）-80μmol/L]亚砷酸钠和[0（对照）-160μg/ml]竹荪多糖＋10μmol/L亚砷酸钠的培养液中暴露24 h,采用四甲基偶氮唑蓝（MTT）法测定细胞存活率。将处于对数生长期的L-02细胞分设对照（培养基）组、亚砷酸钠（10μmol/L）组、竹荪多糖（80μg/ml）组和联合处理（10μmol/L亚砷酸钠＋80μg/ml竹荪多糖）组,暴露24 h后,检测细胞培养液中丙二醛（MDA）、总超氧化物歧化酶（T-SOD）、过氧化氢酶（CAT）、还原性谷胱甘肽（GSH）、细胞色素C（Cyt-C）水平和天冬氨酸蛋白水解酶3（Caspase-3）的活力及B淋巴细胞瘤-2（Bcl-2）蛋白、Bcl-2相关X（Bax）蛋白的表达水平。结果与对照组比较,除2.5和5μmol/L亚砷酸钠染毒组外,10-80μmol/L亚砷酸钠染毒组L-02细胞的存活率均下降,差异有统计学意义（P〈0.05）;且随着亚砷酸钠染毒剂量的升高,L-02细胞的存活率呈下降趋势。与对照组比较,40、80和160μg/ml竹荪多糖联合处理组L-02细胞的存活率均升高,差异有统计学意义（P〈0.05）;其中,以80μg/ml竹荪多糖的拮抗效果最好。与对照组比较,亚砷酸钠组和联合处理组L-02细胞培养液中MDA和Cyt-C的水平升高,T-SOD、CAT及GSH的水平降低,差异均有统计学意义（P〈0.05）;而竹荪多糖组L-02细胞培养液中上述指标均无明显变化。与亚砷酸钠组比较,联合处理组和竹荪多糖组L-02细胞培养液中MDA和Cyt-C的水平降低,T-SOD、CAT及GSH的水平升高,差异均有统计学意义（P〈0.05）。与对照组比较,亚砷酸钠组和联合处理组L-02细胞胞内Caspase-3的活力升高,差异具有统计学意义（P〈0.05）;而竹荪多糖组L-02细胞细胞内Caspase-3的活力无明显变化。与亚砷酸钠组比较,联合处理组和竹荪多糖组L-02细胞内Caspase-3Objective To investigate the antagonism of dictyophora polysaccharide to human normal hepatocytes（L-02 cells）toxicity induced by sodium arsenite. Methods L-02 cells in logarithmic phase were cultured with sodium arsenite at the doses of 0（control）,2.5,5,10,20,40 and 80 μmol/L respectively and then they were co-treated with different concentrations of dictyophora polysaccharide [0（control）,5,10,20,40,80 and 160 μg/ml] combined with 10 μmol/L sodium arsenite for 24 h respectively. The cell livability was determined by MTT assay. L-02 cells in logarithmic phase were cultured with culture medium（control group）,10 μmol/L sodium arsenite（sodium arsenite group）,80 μg/ml dictyophora polysaccharide（dictyophora polysaccharide group） and 10 μmol/L sodium arsenite ＋80 μg/ml dictyophora polysaccharide（co-treated group） for 24 h respectively. Malondialdehyde（MDA）,total superoxide dismutase（T-SOD）,catalase（CAT）,glutathione（GSH） and cytochrome C（Cyt-C） in culture medium of each group were determined. The protein levels of aspartic protease 3（caspase-3）,B-cell lymphoma 2（Bcl-2） protein and Bcl-2-associated X（Bax） protein of L-02 cells in each group were determined. Results Compared with the control group,the livabilities of L-02 cells were significantly decreased in sodium arsenite groups except 2.5and 5 μmol/L sodium arsenite groups; The livability tendency of L-02 cells was decreased as the dose of sodium arsenite increased. Compared with the control group,the livabilities of L-02 cells were significantly increased in co-treated groups including 40,80 and 160 μg/ml dictyophora polysaccharide,especially at the concentration of 80 μg/ml. Compared with the control group,the levels of MDA and Cyt-C were increased and the levels of T-SOD,CAT and GSH were decreased in L-02 cells culture medium in sodium arsenite and co-treated groups（P〈0.05）;There was no significantly difference between control and dictyophora polysaccharide group in the indexes above. Comp

关 键 词：竹荪多糖 亚砷酸钠 线粒体 细胞凋亡 

分 类 号：R994.6[医药卫生—毒理学]