谷玉祥, 万煜, 薛鹏. 碘乙腈致三种人源性肝细胞氧化应激作用的研究[J]. 环境与职业医学, 2016, 33(12): 1127-1132. DOI: 10.13213/j.cnki.jeom.2016.16541
引用本文: 谷玉祥, 万煜, 薛鹏. 碘乙腈致三种人源性肝细胞氧化应激作用的研究[J]. 环境与职业医学, 2016, 33(12): 1127-1132. DOI: 10.13213/j.cnki.jeom.2016.16541
GU Yu-xiang, WAN Yu, XUE Peng. Iodoacetonitrile Induced Oxidative Stress in Three Types of Human Hepatocytes[J]. Journal of Environmental and Occupational Medicine, 2016, 33(12): 1127-1132. DOI: 10.13213/j.cnki.jeom.2016.16541
Citation: GU Yu-xiang, WAN Yu, XUE Peng. Iodoacetonitrile Induced Oxidative Stress in Three Types of Human Hepatocytes[J]. Journal of Environmental and Occupational Medicine, 2016, 33(12): 1127-1132. DOI: 10.13213/j.cnki.jeom.2016.16541

碘乙腈致三种人源性肝细胞氧化应激作用的研究

Iodoacetonitrile Induced Oxidative Stress in Three Types of Human Hepatocytes

  • 摘要: 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0~100 μmol/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

    ①50 μmol/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;100 μmol/L组,3种肝细胞大量死亡。②HepG2细胞存活力在100 μmol/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20 μmol/L时,Chang liver细胞存活力明显降低(均P<0.05);LO2细胞存活力在10 μmol/L组明显上升(P<0.05),在50和100 μmol/L组明显下降(均P<0.05)。③在100 μmol/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。④ARE报告基因活性随着IAN浓度的增加而上升(r=0.944,P<0.05)。在5 μmol/L和10 μmol/L组,ARE报告基因的活性明显上升(均P<0.05)。⑤在20 μmol/L组,3种肝细胞内SOD活性明显下降(均P<0.05);而在1 μmol/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。

     

    Abstract: Objective

    To study the characteristics and mechanism of toxic effects of iodoacetonitrile (IAN) on human hepatocytes,and select suitable cells for examining the toxic effects of environmental pollutants.

    Methods

    After IAN exposure at various concentrations,human liver cancer cells (HepG2) and human normal liver cells (Chang Liver and LO2) were observed for the morphological changes under microscope and detected for cell viability with Cell Counting Kit-8(CCK-8) and intracellular level of reactive oxygen species (ROS),antioxidant response element (ARE) luciferase reporter gene activity,glutathione (GSH) content,and superoxide dismutase (SOD) activity simultaneously.

    Results

    ①The morphology of the three types of human hepatocytes in the 50 μmol/L groups showed round and shrinking cells with fuzzy cell edges and disappeared intercellular junctions;the 100 μmol/L groups showed massive apoptosis.② The viability of HepG2 decreased significantly in the 100 μmol/L group (P<0.05).The viability of Chang liver decreased with higher concentrations of IAN (r=-0.974,P<0.05) and compared with control group,the viability of Chang liver decreased significantly when the concentration of IAN was over 20 μmol/L (all Ps<0.05).The viability of LO2 increased in the 10 μmol/L group (P<0.05) while decreased in the 50 μmol/L and 100 μmol/L groups significantly (both Ps<0.05).③ In the 100 μmol/L groups,the levels of ROS of the three types of human hepatocytes were higher than that of the control group (all Ps<0.05).Among them,the intracellular ROS of HepG2 increased slightly.④ The ARE reporter gene activity varied depending on the concentration of IAN (r=0.944,P<0.05) and increased significantly in the 5 μmol/L groups and the 10 μmol/L groups (both Ps<0.05).⑤ The activities of SOD in the three types of human hepatocytes all decreased (all Ps<0.05) in the 20 μmol/L group,but the SOD in Chang liver cells increased (P<0.05) in the 1 μmol/L group.⑥ The changes of GSH in three human hepatocytes were not significant (all Ps>0.05).

    Conclusion

    The cytotoxicity of IAN on the three types of human hepatocytes can be caused by oxidative stress.Compared to HepG2,Chang Liver and LO2 are more sensitive to the toxicity of IAN and suitable to be the cell models for studying toxic effects of environmental pollutants.

     

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