Study on the effects of endocrine disrupter on an organism using the indicators in apoptosis

近年来，研究表明，环境中的内分泌干扰物（对羟基苯甲酸酯、六六六、滴滴涕、二恶英、壬基酚、阿特拉津、双酚A、DEHP等）有直接影响心理或神经通路的作用。我们建立了PC-12细胞培养体系，以研究其凋亡机制. PC 12细胞具有神经元样细胞的功能，该系统具有凋亡的特征，本研究采用Western印迹分析和RT-PCR方法研究了γ-六氯环己烷对PC 12细胞凋亡的影响。高浓度的γ-HCH对细胞活力和凋亡无影响，但在低浓度时，细胞DNA断裂程度略有增加，凋亡因子Bax、Bad、细胞色素c和caspase-3有增加的趋势。然而，这些变化没有统计学意义。结论是，γ-HCH在PC 12细胞系系统中不影响细胞凋亡，尽管据报道γ-HCH在其他细胞系中可诱导细胞凋亡。为了解对羟基苯甲酸酯与胚胎和婴儿分化的关系，研究了对羟基苯甲酸酯对血清剥夺诱导的PC 12细胞凋亡的影响。采用ELISA和RT-PCR方法检测凋亡相关因子caspase-3和Bcl-2家族的表达。其结果是，我们发现，对羟基苯甲酸甲酯增强细胞凋亡，和增强被认为是由增加坏，促凋亡蛋白，虽然对羟基苯甲酸丁酯不影响细胞凋亡。

Recently it is suggested that the endocrine disrupters (parabens, g-hexachlorocyclohexane (HCH), DDT, dioxin, nonylphenol, atrazine, bisphenol-A, DEHP etc.) have effect that affects directly to the psychological or neurological pathway. We developed culture system of PC 12 cells to study about the mechanism of apoptosis. The PC12 cell acts like a neuronal cell and this system shows some indicators of apoptosis.In this study, the effects of γ-hexachlorocyclohexane on apoptosis in PC12 cells were investigated using western blotting analysis and RT-PCR method. Although γ-HCH at high concentration did not effect on cell viability and apoptosis, DNA fragmentation was slightly enhanced, and apoptotic factors; Bax, Bad, cytochrome c and caspase-3 showed tendency of increase by addition of low γ-HCH to the cell medium. However these changes were not statistically significant. It was concluded that γ-HCH did not affect on apoptosis in the PC12 cell line system, although γ-HCH has been reported to induce apoptosis in other cell lines.The alkyl esters of p-hydroxybenzoic acid (parabens) were also investigated. To understand the relationship between parabens and differentiation in embryos and infants, the effects of parabens on apoptosis induced by serum deprivation in PC12 cells were investigated. In addition, apoptosis-related factors such as caspase-3 and the Bcl-2 family were assayed by ELISA and RT-PCR methods. As a result, we found that methylparaben enhanced apoptosis, and the enhancement was suggested to be caused by an increase of Bad, a proapoptotic protein, although butylparaben did not affect apoptosis.