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[目的]了解碘乙腈(IAN)对多种人源性肝细胞的毒性作用特征,初步探索其毒性作用机制,并选取适合用于研究环境污染物毒性效应的细胞模型。[方法]将人肝癌细胞(HepG2)、人正常肝细胞(Chang liver和LO2)行0~100μmol/L不同浓度的IAN染毒,镜下观察各染毒剂量下细胞的形态结构变化,并用细胞增殖毒性检测试剂盒(CCK-8)检测细胞存活力,同时测定细胞内活性氧(ROS)水平、抗氧化反应元件(ARE)报告基因活性、谷胱甘肽(GSH)水平、超氧化物歧化酶(SOD)活性。[结果]150μmol/L染毒组,3种肝细胞逐渐变圆皱缩、边界模糊不清、细胞间连接消失;100μmol/L组,3种肝细胞大量死亡。2 HepG2细胞存活力在100μmol/L组明显下降(P<0.05);Chang liver细胞存活力随着IAN浓度的升高而降低(r=-0.974,P<0.05),当IAN浓度大于20μmol/L时,Chang liver细胞存活力明显降低(均P<0.05);LO2细胞存活力在10μmol/L组明显上升(P<0.05),在50和100μmol/L组明显下降(均P<0.05)。3在100μmol/L组,3种肝细胞内ROS水平明显上升(均P<0.05),其中HepG2上升幅度较小。4 ARE报告基因活性随着IAN浓度的增加而上升(r=0.944,P<0.05)。在5μmol/L和10μmol/L组,ARE报告基因的活性明显上升(均P<0.05)。5在20μmol/L组,3种肝细胞内SOD活性明显下降(均P<0.05);而在1μmol/L组,Chang liver细胞内SOD活性明显上升(P<0.05)。6各浓度组3种肝细胞的细胞内GSH水平变化,均无统计学意义(均P>0.05)。[结论]IAN可在3种人源性肝细胞内诱导氧化应激,进而造成细胞损伤。与HepG2相比,Chang liver和LO2对IAN诱导的细胞毒性和氧化应激效应更敏感,更适合作为研究环境污染物毒性效应的细胞模型。
[Objective] To understand the toxicity of iodine-acetonitrile (IAN) on a variety of human hepatocytes and to explore the mechanism of its toxicity, and to select a suitable cell model for studying the toxic effects of environmental pollutants. [Method] The human hepatocellular carcinoma cells (HepG2) and human normal liver cells (Chang liver and LO2) were exposed to 0 ~ 100μmol / L different concentrations of IAN. The morphological changes of the cells were observed under microscope. The cell viability was measured by proliferation assay kit (CCK-8). The level of reactive oxygen species (ROS), the activity of reporter gene, glutathione (GSH), superoxide dismutase Enzyme (SOD) activity. [Results] The three kinds of hepatocytes gradually became round and wrinkled in the 150 μmol / L group, the boundaries were blurred and the intercellular connections disappeared. A large number of three kinds of hepatocytes died in the group of 100 μmol / L. The viability of HepG2 cells in 100 μmol / L group decreased significantly (P <0.05). The viability of Chang liver cells decreased with the increase of IAN concentration (r = -0.974, P <0.05). When IAN concentration was more than 20 μmol / L (P <0.05). The viability of LO2 cells was significantly increased in 10μmol / L group (P <0.05), but significantly decreased in 50μmol / 100μmol / L group (all P <0.05). 3 in 100μmol / L group, 3 kinds of hepatocytes ROS levels were significantly increased (all P <0.05), of which HepG2 increased less. The 4 ARE reporter gene activity increased with increasing IAN concentration (r = 0.944, P <0.05). In 5μmol / L and 10μmol / L groups, the activity of ARE reporter gene was significantly increased (all P <0.05). 5 in 20 μmol / L group, the activity of SOD in 3 kinds of hepatocytes decreased significantly (all P <0.05); while in 1 μmol / L group, the activity of SOD in Chang liver cells increased obviously (P <0.05). There was no significant difference in intracellular GSH levels among three kinds of hepatocytes in each concentration group (all P> 0.05). [Conclusion] IAN can induce oxidative stress in 3 kinds of human hepatocytes, leading to cell injury. Compared with HepG2, Chang liver and LO2 are more sensitive to IAN-induced cytotoxicity and oxidative stress, which is more suitable as a cellular model for studying the toxic effects of environmental pollutants.