亚硫酸钠引起肝细胞内活性氧增高对p53蛋白表达的影响  

Effect of increased reactive oxygen species in hepatocytes induced by sodium suifite on p53 expression

作　　者：雷佩玉[1] 刘鑫[2] 赵春燕[1] 仪慧兰[2] 王幼萍[1] 梁瑞峰[1] 白剑英[1] 

机构地区：[1]山西医科大学公共卫生学院环境卫生学教研室,太原030001 [2]山西大学生命科学院,太原030006

出　　处：《解剖学杂志》2012年第4期420-422,F0003,共4页Chinese Journal of Anatomy

基　　金：国家自然科学基金（30972445）;山西省自然科学基金（2007011109）第1作者E-mail：[e-mail]

摘　　要：目的：探讨亚硫酸钠引起肝细胞内活性氧水平变化与p53蛋白表达之间的关系。方法：将人正常二倍体肝细胞株HL-7702培养于含10％小牛血清的DMEM培养基中，待生长至适宜密度时用不同浓度亚硫酸钠染毒4h。采用Fluo-3AM钙离子荧光指示剂法测定肝细胞内Ca^2＋。水平变化，二氯荧光黄双乙酸酯荧光标记法测定肝细胞内活性氧水平，蛋白杂交法和免疫细胞化学法检测肝细胞内p53蛋白表达情况，RT-PCR法检测肝细胞内p53基因mRNA表达水平的变化。结果：一定浓度的亚硫酸钠可引起肝细胞内Ca^2＋水平增加，并伴随着活性氧水平增加和肝细胞内p53蛋白表达水平明显增加，但p53基因mRNA表达水平无明显变化。结论：亚硫酸钠可通过增加肝细胞内Ca^2＋水平引起肝细胞内活性氧水平和p53蛋白水平增加，这种影响可能发生在蛋白质翻译或翻译后水平。Objective： To study the possible relationship between reactive oxygen species （ROS） production and p53 protein expression of hepatocytes induced by sodium sulfite. Methods： The HL-7702 hepatocytes were cultured in DMEM medium containing 10% new born bovine serum. After 4h, intracellular calcium level was demonstrated using Fluo-3 acetomethoxyester and the generation of ROS was probed with 2＇, 7＇-dichlorofluorescein diacetate （DCFH DA） fluorescence. The expression of p53 protein was detected by immunohistochemistry and Western blotting. The mRNA expression of p53 was demonstrated by RT-PCR. Results： Na2 SO2 increased Ca^2＋ level inside hepatocytes with an increase of ROS. The expression of p53 protein was also promoted by Na2SO3 treatment, and no significant changes in mRNA level of p53 were found. Conclusion： Na2SO3 can promote ROS production with an increase in p53 protein expression, which may be due to the increase of Ca^2＋ , and result in liver injury.

关 键 词：亚硫酸钠 钙离子浓度 活性氧 P53 

分 类 号：R735.34[医药卫生—肿瘤]