Effect of disturbing endocrine drug, DEHP, on microcyst formation in the epithelial cells.

液泡H ^+-ATP酶（V-ATP酶）定位于中央液泡系统的细胞器如溶酶体、被膜囊泡和高尔基体中，在维持这些细胞器的酸性环境中起重要作用。研究了邻苯二甲酸二（2-乙基己基）酯（DEHP）对小鼠肝溶酶体的影响。小鼠口服给药2 - 3周后，通过免疫细胞化学分析确定，空泡H ^+-ATP酶减少。当小鼠随后被喂食正常饮食1周时，V-ATP酶水平恢复到正常值。北方印迹分析显示，随着DEHP处理，V-ATP酶A亚基mRNA逐渐降低。酶细胞化学染色显示酸性磷酸酶存在于正常动物和DEHP处理动物的溶酶体和晚期自噬体中。但DEHP处理后，含酸性磷酸酶的晚期自噬体数量明显增加。这些结果表明，DEHP导致肝脏溶酶体室中V-ATP酶显著降低，并且对DEHP的影响是可逆的，DEHP对蛋白质表达的影响可能在转录水平上发挥作用。因此，在本研究中，我们报告DEHP处理导致肝脏溶酶体室中V-ATP酶亚基A减少，这可能是由于无法降解多余的细胞器。

Vacuolar H^+-ATPase(V-ATPase) is localized in organelles of the central vacuolar system such as lysosomes, coated vesicles, and the Golgi apparatus, and it plays an important role in maintaining the acidic enviroment in these compartments. We investigated the effects of di(2-ethylhexyl) phthalate (DEHP) on mouse liver lysosomes. After 2-3 weeks of oral administration in mice, a reduction in vacuolar H^+-ATPase, as determined by immunocytochemical analysis. When the mice were subsequently fed a normal diet for 1 week, V-ATPase, levels recovered to normal values. According to Northern blot analysis, V-ATPase subunit A mRNA decreased gradually with DEHP treatment. Enzyme cytochemical staining showed acid phosphatase to be present in lysosomes and late autophagosomes in normal animals as well as in DEHP-treated animals. But the number of late autophagosomes containing acid phosphatase increased clearly after DEHP treatment. These results suggest that, DEHP causes marked V-ATPase reduction in the liver lysosomal compartment and the effect to DEHP is reversible, and the effect of DEHP on protein expression is likely to be exerted at the transcriptional level. Thus in this study, we report that DEHP treatment causes a reduction in V-ATPase subunit A in the liver lysosomal compartment, which may accent for the inability to degrade excess cell organelles.