亚硫酸钠对人正常二倍体肝细胞内二酰甘油酰基转移酶1和载脂蛋白E mRNA和蛋白表达的影响  

Effects of sodium sulfite on mRNA and protein expression of diacylglycerol acyltrartsferase 1 and apolipoprotein E of HL-7702 cells

作　　者：王幼萍[1] 白剑英[1] 雷佩玉[1] 闫丹丹[1] 梁瑞峰[1] 赵五红[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,山西太原030001

出　　处：《环境与健康杂志》2013年第1期5-8,F0003,共5页Journal of Environment and Health

基　　金：国家自然科学基金(30972445);山西省回国留学人员科研资助项目(2009-53);山西高校科技研究开发项目(200613014)

摘　　要：目的通过研究亚硫酸钠对人正常二倍体肝(HL-7702)细胞内二酰甘油酰基转移酶1(DGAT1)和载脂蛋白E(apoE)mRNA和蛋白表达的影响,探讨亚硫酸钠诱发肝细胞脂肪积累效应及作用途径。方法将处于对数生长期的HL-7702细胞株分别暴露于终浓度为0(阴性对照)、10、2.5、0.5、0.1 mmol/L的亚硫酸钠溶液,并设立阳性对照(1.0 mmol/L油酸)组。分别于染毒24、48 h时,采用油红O染色法观察肝细胞内脂滴沉积,采用免疫沉淀和Western Blot法检测细胞内apoE和DGAT1蛋白的表达水平,采用qRT-PCR法检测肝细胞内apoE和DGAT1 mRNA的表达水平。结果与阴性对照组比较,仅10 mmol/L亚硫酸钠染毒24 h时HL-7702肝细胞内DGAT1 mRNA表达水平明显增加,差异有统计学意义(P<0.05)。与阴性对照组比较,染毒24h、48 h时,10 mmol/L亚硫酸钠染毒组HL-7702肝细胞内DGAT1蛋白表达水平以及各浓度亚硫酸钠染毒组HL-7702肝细胞内apoE蛋白表达水平均明显增加,差异有统计学意义(P<0.05)。阳性对照组可见明显的脂滴沉积,但亚硫酸钠染毒组均未见明显脂滴。结论一定剂量的亚硫酸钠可引起肝细胞内DGAT1、apoE蛋白表达增高,导致肝细胞内甘油三酯分泌增加,从而引起肝细胞的脂肪蓄积。Objective To study the effect of sodium sulfite on mRNA and protein expression of diacylglycerol acyltrartsferase 1 and apolipoprotein E of HL-7702, and discuss the mechanism of sodium sulfite induced hepatic fat accumulation. Methods The HL-7702 cell line in logarithmic growth phase were treated with 0,10,2.5,0.5,0.1 mmol/L Na2SO3 and positive control group was set, for 12 and 24 h. Oil red O was used to detect bullous fat deposition in hepatocytes, immunoprecipitation and Western Blot was used to detect the expression of apolipoprotein （apoE） and diacylglycerol acyhransferase 1 （DGAT1） protein in hepatocytes,qRT-PCR was used to detect the mRNA level of apoE and DGAT1 in hepatocytes. Results Compared with the control, DGAT1 mRNA level in HL-7702 hepatocyte increased significantly in 10 mmol/L sodium sulfite group exposed for 24 h （P〈0.05）. Compared with control, the expression of DGAT1 protein increased significantly in 10 mmol/L sodium sulfite group exposed for 24 and 48 h, and the expression of apoE protein increased significantly in each sodium sulfite group exposed for 24 and 48 h（P〈0.05）. Positive control showed obvious lipid droplets, but Na2SO3-treated groups did not show significant lipid droplets. Conclusion Certain dose of sodium sulfite can promote the expression of DGAT1 and apoE protein in hepatocytes and the secretion of triglyceride, and induce lipid accumulation in hepatocytes.

关 键 词：亚硫酸钠 载脂蛋白E 二酰甘油酰基转移酶1 免疫沉淀 

分 类 号：R994.6[医药卫生—毒理学]