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

近年来，人们认为内分泌干扰物（对羟基苯甲酸酯、g-六氯环己烷、DDT、二恶英、壬基酚、阿特拉津、双酚a、DEHP等）对心理或神经通路有直接影响。我们建立了pc12细胞培养体系，研究其凋亡机制。PC12细胞的行为类似于神经元细胞，该系统显示出一些凋亡的指标。本研究采用western blotting和RT-PCR方法研究γ-六氯环己烷对PC12细胞凋亡的影响。虽然高浓度γ-HCH对细胞活力和凋亡没有影响，但DNA片段化略有增强，凋亡因子；添加低γ-HCH后，Bax、Bad、细胞色素c和caspase-3均有升高的趋势。然而，这些变化在统计学上并不显著。结果表明，γ-HCH不影响PC12细胞系系统的凋亡，尽管γ-HCH在其他细胞系中有诱导凋亡的报道。对羟基苯甲酸（对羟基苯甲酸酯）的烷基酯也进行了研究。为了了解对羟基苯甲酸酯与胚胎和婴儿分化的关系，我们研究了对羟基苯甲酸酯对血清剥夺诱导的PC12细胞凋亡的影响。此外，采用ELISA和RT-PCR方法检测caspase-3、Bcl-2家族等凋亡相关因子。结果，我们发现对羟基苯甲酸甲酯促进了细胞凋亡，而这种增强可能是由促凋亡蛋白Bad的增加引起的，尽管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.