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白剑英, 雷佩玉, 王幼萍, 闫丹丹, 梁瑞峰, 王祝欣. 四氯化碳对HL-7702肝细胞甘油三酯含量影响[J]. 中国公共卫生, 2014, 30(1): 86-88. DOI: 10.11847/zgggws2014-30-01-26
引用本文: 白剑英, 雷佩玉, 王幼萍, 闫丹丹, 梁瑞峰, 王祝欣. 四氯化碳对HL-7702肝细胞甘油三酯含量影响[J]. 中国公共卫生, 2014, 30(1): 86-88. DOI: 10.11847/zgggws2014-30-01-26
BAI Jian-ying, LEI Pei-yu, WANG You-ping.et al, . Effects of carbon tetrachloride on triglyceride content in HL-7702 liver cells[J]. Chinese Journal of Public Health, 2014, 30(1): 86-88. DOI: 10.11847/zgggws2014-30-01-26
Citation: BAI Jian-ying, LEI Pei-yu, WANG You-ping.et al, . Effects of carbon tetrachloride on triglyceride content in HL-7702 liver cells[J]. Chinese Journal of Public Health, 2014, 30(1): 86-88. DOI: 10.11847/zgggws2014-30-01-26

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

  • 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。

     

    Abstract: Objective To observe the effects of carbon tetrachloride (CCl4)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 CCl4 and positive control (1 mmol/L oleic acid).After 24 and 48 hours,TG content was detected with glycerol-phosphate oxidase-peroxidase (GPO-POD)assay.Western blott was used to detect secretion of VLDL and protein expressions of microsomal triglyceride transfer protein (MTP),diacylglycerol acyltransferase 2(CDGAT2),and carnitine palmitoyl transferase IA(CPTIA)in hepatocytes.ApoB100 secretion was detected with enzyme-linked immunosorbent assay (ELISA).Results After 24 hours' exposure,CCl4 increased TG level significantly(3.2353±1.1168 mmol/g prot for CCl4 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 CCl4 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 CCl4 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 CCl4 may increase TG level in hepatocytes by increasing DGAT2 expression,and enhance the secretion of ApoB100 by increasing MTP expression in normal hepatocytes.

     

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