空气负氧离子抑制肝癌细胞的实验研究(精选).docVIP

空气负氧离子抑制肝癌细胞的实验研究 Study of Negative oxygen ions inhibiting liver cancer cells in vitro 　　TAO Ming-zhang1，LI Hui2,CHEN Shao-zhou3,LI Hai-tao4 1.China on Negative Air Ions and Ozone Reseach Society，Guangzhou 510300，China;2. Center of Respiration and Sleep，Nanfang Hospital Affiliated Nanfang Medical College，Guangzhou 510516，China;3. Department of Psychiatry，Hongkong Queen Mary Hospital，Hongkong 999077，China;4. Department of Neurology，First Hospital Affiliated SUN YAT-SEN University，Guangzhou 510080，China 【Abstract】Objective：To investigate the affect on the biologic activity of hepatoma Hepa1-6 cell line of negative oxygen ions in vitro. Methods Dividing the cell into three groups: negative control group, air control 　　group and treatment group，randomly.Dividing the treatment group into 1h、2h and 4h group according to action time of negative oxygen ions, all the cells in each group were detected apoptosis and cell cycle with flow cytometry method, the phospho- P38MAPK expression was detected. 　　by Westernblot assay. Results：The apoptosis rate increased, the proportion of S stage cell decreased and the expression of phospho- P38MAPK increase obviously vs negative and air control group, the difference show significance P 0.05 . 　　Conclusion：The negative oxygen ions can induce hepatoma Hepa1-6 cell apoptosis through arresting the cell cycle, such effect was mediated by P38MAPK pathway. 　　Key words: Negative oxygen ions; Negative air ions; Hepatoma; P38MAPK 　　被誉为空气维生素的空气负氧离子对人体健康十分有益。研究表明，它能有效调整大脑皮层的功能，有镇静、催眠及降血压作用;它能使支气管平滑肌松弛，解除其痉挛，有效缓解哮喘;它还可以降低血脂水平，提高机体免疫力及抗肿瘤作用等[1-4]。目前大多数研究集中于流行病学方面的研究，而对于抗肿瘤和及其作用机理的研究很少。本实验就空气负氧离子对体外肝癌细胞的影响进行了系列研究。 　　1. 材料与方法 　　1.1 材料 　　肝癌细胞株Hepa1-6 上海艾研生物科技有限公司 ;碘化丙啶 美国Sigma 公司 ;TUNEL细胞凋亡检测试剂盒 德国Boehringer Mannheim公司 ;鼠抗人单克隆抗体P38MAPK和P38MAPK 特异性抑制剂SB203580 美国On-cogene公司 ，SB203580 使用终浓度为50μmol/L;森肽基生态级负氧离子生成机 济南新活电器公司 ;Model IC-2000型空气负离子检测器 瑞典，空气负离子检测装置 。 　　1.2 方法 　　1.2.1 实验分组 　　把处于对数生长期的肝癌细胞Hepa1-6随机分成3组, 即空白对照组、空气对照组及负氧离子治疗组。治疗组根据空气负离子作用时间分成1h组、