四氯化碳对HL-7702肝细胞甘油三酯含量影响  

Effects of carbon tetrachloride on triglyceride content in HL-7702 liver cells

作　　者：白剑英[1] 雷佩玉[1] 王幼萍[1] 闫丹丹[1] 梁瑞峰[1] 王祝欣[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：《中国公共卫生》2014年第1期86-88,共3页Chinese Journal of Public Health

基　　金：山西省回国留学人员科研资助(2009-53);山西高校科技研究开发资助(200613014)

摘　　要：目的探讨四氯化碳(CCl4)对人正常二倍体HL-7702肝细胞内甘油三酯含量(TG)和极低密度脂蛋白(VLDL)分泌影响。方法实验设阴性对照组、CCl4组(10 mmol/L)和阳性对照组(1 mmol/L油酸),染毒结束后检测TG含量,采用磷钨酸盐沉淀和蛋白印迹法检测VLDL含量,用蛋白印记法检测甘油三酯转移蛋白(MTP),二脂酰甘油酰基转移酶2(DGAT2)和肉碱酰基转移酶(CPT1A)蛋白表达水平,用酶联免疫法检测ApoB100含量。结果染毒24 h后,CCl4组细胞内TG含量[(3.235 3±1.116 8)mmol/g prot]明显高于阴性对照组[(1.221 8±0.527 2)mmol/g prot](P<0.05);VLDL分泌水平无明显改变;细胞内MTP、DGAT2蛋白表达量明显增加;染毒24、48 h后,CCl4处理组肝细胞培养上清中ApoB100蛋白含量分别为231.79、263.33 ng/mL,较阴性对照组(135.12、143.59 ng/mL)明显增加(P<0.05)。结论四氯化碳可能通过促进DGAT2蛋白表达而引起肝细胞内TG含量增加,或通过MTP途径促进ApoB100分泌,但对VLDL分泌无影响。Objective To observe the effects of carbon tetrachloride （ CCI4 ） on triglyceride （TG） level and the change of very low density lipoprotein （VLDL） secretion in hepatocytes. Methods Human diploid liver cells （ HL- 7702）, cultured in Dulbecco＇ s modified Eagle＇ s medium （DMEM） containing 10% fetal bovine serum, were divided into 3 groups：negative control, 10 mmol/L CCI4 and positive control （ 1 mmol/L oleic acid）. After 24 and 48 hours, TG content was detected with glycerol-phosphate oxidaseperoxidase （GPO-POD） assay. Western blott was used to detect se cretion of VLDL and protein expressions of microsomal triglyceride transfer protein （MTP）, diacylglycerol acyltrans ferase 2 （CDGAT2）, and carnitine palmitoyl transferase IA （CPTIA）in hepatocytes. ApoB 100 secretion was detected with enzyme-linked immunosorbent assay （ELISA）. Results After 24 hours＇ exposure,CClg increased TG level significant ly（3. 2353 ± 1. 1168 mmol/g prot for CC14 group vs 1. 2218 ± 0. 5272 mmol/g prot for negative control group ）, （P 〈 0. 05 ）. VLDL secretion was not changed. After exposure of 24 hours and 48 hours, the protein levels of MTP and DGAT2 in the CCI4 treated group were higher than those of negative control group;the expression of CPT1 in 48 hours exposure group was significantly lower than that of the control. The ApoB100 protein levels in culture medium of CC14 treated groups （231.79,263.33 ng/mL for 24 hours and 48 hours）were significantly higher in comparison with the negative control group （135.12,143.59 ng/mL）（P 〈 0. 05）. Conclusion CCI4 may increase TG level in hepatocytes by increasing DGAT2 ex- pression, and enhance the secretion of ApoB100 by increasing MTP expression in normal hepatocytes.

关 键 词：四氯化碳 甘油三酯(TG) 极低密度脂蛋白(VLDL) 甘油三酯转移蛋白(MTP) 二脂酰甘油酰基转移酶2(DGAT2) 载脂蛋白B100 

分 类 号：R12[医药卫生—环境卫生学]