α7nAChR在氯化甲基汞神经毒性中的作用

为探讨环境染污染物氯化甲基汞(methylmercury chloride,Me Hg Cl)的神经毒性作用机制,采用MTT、免疫细胞荧光、dot Blot、qRT-PCR等实验技术检测Me Hg Cl对PC12细胞活性及其α7亚型烟碱型胆碱能受体(α7 nicotinic acetylcholine receptor,α7n AChR)蛋白和mRNA表达水平的影响。结果显示Me Hg Cl抑制PC12细胞活性,显著降低α7n AChR的蛋白和mRNA的表达水平,呈浓度依赖性。上述结果表明,Me Hg Cl对神经细胞毒性作用可能与抑制α7n AChR表达有关。     

In vitro evaluation of inorganic and methyl mercury mediated cytotoxic effect on neural cells derived from different animal species

To extend the current understanding of the mercury-mediated cytotoxic effect,five neural cell lines established from different animal species were comparatively analyzed using three different endpoint bioassays:thiazolyl blue tetrazolium bromide,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay(MTT),neutral red uptake assay(NRU),and Coomassie blue assay(CB).Following a 24-hr exposure to selected concentrations of mercury chloride(HgCl_2) and methylmercury(Ⅱ) chloride(MeHgCl),the cytotoxic effect on test cells was characterized by comparing their 50%inhibition concentration(IC_(50)) values.Experimental results indicated that both these forms of mercury were toxic to all the neural cells,but at very different degrees.The IC_(50)values of MeHgCl among these cell lines ranged from 1.15±0.22 to 10.31 ± 0.70 μmol/L while the IC_(50) values for HgCl_2 were much higher,ranging from 6.44 ± 0.36 to 160.97±19.63 μmol/L,indicating the more toxic nature of MeHgCl.The IC_(50) ratio between HgCl_2and MeHgCl ranged from 1.75 to 96.0,which confirms that organic mercury is much more toxic to these neural cells than inorganic mercury.Among these cell lines,HGST-BR and TriG44 derived from marine sea turtles showed a significantly high tolerance to HgCl_2 as compared to the three mammalian neural cells.Among these neural cells,SK-N-SH represented the most sensitive cells to both chemical forms of mercury.