致肝损伤毒物四氯化碳体外染毒剂量的选择  

Selection of the appropriate exposure dose of carbon tetrachloride for inducing the liver injury in vitro

作　　者：张继东[1] 张健新[2] 王幼萍[2] 白剑英[2] 

机构地区：[1]山西省肿瘤医院放疗腹盆二病区,太原030013 [2]山西医科大学公共卫生学院环境卫生学教研窒

出　　处：《山西医科大学学报》2013年第4期263-266,共4页Journal of Shanxi Medical University

基　　金：山西省自然科学基金资助项目(2007011109);山西高校科技研究开发项目(200613014)

摘　　要：目的探讨四氯化碳(CCl4)致肝脏损伤体外实验模型的染毒剂量。方法以不同浓度CCl4作用于正常人二倍体肝细胞HL-7702细胞,使其终浓度分别为0(阴性对照),5,10,20,30 mmol/L和40 mmol/L。采用CCK8法检测不同浓度CCl4对肝细胞活性的影响,采用生化分析法检测染毒后肝细胞培养上清中谷草转氨酶(AST),谷丙转氨酶(ALT),乳酸脱氢酶(LDH),碱性磷酸酶(AKP)的水平。结果①染毒24 h后,肝细胞活性呈剂量依赖性降低,其中20-40 mmol/L CCl4对肝细胞活性有明显抑制作用,与对照组相比有统计学差异(P<0.05)。②染毒24 h后,随着CCl4染毒剂量的增加,各染毒组肝细胞培养上清中AST的活性有逐渐升高的趋势,其中40 mmol/L CCl4染毒组与阴性对照组比较有统计学差异(P<0.05);各染毒组肝细胞培养上清中ALT的活性逐渐增加,其中10-30 mmol/L CCl4染毒组与阴性对照组相比显著升高(P<0.05);各染毒组肝细胞培养上清中AKP和LDH的活性与阴性对照组相比较差异无统计学意义(P>0.05)。结论 10 mmol/L CCl4在对HL-7702肝细胞活性无明显影响的情况即可引起肝细胞培养上清中ALT的活性升高,对肝细胞膜造成一定的损伤,可作为其肝损伤机制研究的体外染毒剂量。Abstract： Objective To explore an appropriate exposure dose of carbon tetrachloride （CC14 ） for inducing liver injury models in vitro. Methods The normal human diploid strains HL-7702 cells were cultured in DMEM medium containing 10% fetal bovine serum （ FBS）, and treated by different concentrations of CC14 at the final concentrations of 0,5,10,20,30 mmol/L and 40 mmol/L, respective- ly. CCK8 assay was used to detect the cell viability, and the biochemical assay was used to detect levels of aspartate aminotransferase （ AST）, alanine aminotransferase （ ALT）, lactate dehydrogenase （ LDH ）, alkaline phosphatase （AKP） in the culture supernatant of liver cells after exposure for 24 h. Results ①After exposure to CCl4,the cell viability was inhibited in a dose-dependent manner, especial- ly 20 - 40 mmol/L CC14 （ vs negative controls, P 〈 0.05 ）. ②ith the increase of the dose of CCI4, AST activity in the culture superna- tant increased gradually. AST activity in 40 mmol/L CC14 group was significantly higher than that in negative control group （ P 〈 0.05 ）. After CCl4 exposure for 24 h, ALT activity in the culture supernatant increased significantly, especially 10 -30 mmol/L （vs negative control group,P 〈 0.05 ）. After CCl4 exposure for 24 h, LDH and AKP activity in the culture supernatant increased gradually, but no statistical significance was found compared with negative controls. Conclusion The 10 mmol/L CC14 can not inhibit the cell viability of HL-7702 liver cells ,but can increase the ALT activity in the culture supernatant of liver cells, which means a damage of the liver cells membrane. So it may be an appropriate dose for in vitro study on CCl4 toxicity.

关 键 词：四氯化碳(CCl4) 谷草转氨酶(AST) 谷丙转氨酶(ALT) CCK8法 HL-7702肝细胞 

分 类 号：R332[医药卫生—人体生理学]