DEHP Effects On Male Estrogen Effect And Apoptosis Of Liver Cells Rana China

In recent years, a large number of synthetic compounds have been released into the nature along with the rapid development of industry and agriculture.Many of compounds can disturb the normal growth and development in wild animal. Phthalic acid esters (PAEs) as industrial products are used universally in our daily life which can induce effect of estrogen. Di2-Ethyl Hexyl Phthalate is one of the PAEs. The report about whether it can induce estrogenic effect or apoptosis in hepatic cells of amphibians hasn’t been studied. So we take R. chensinensis as experimental anmial to understand the estrogenic effect and to investigate the effect of hepatocyte apoptosis of DEHP. The aim of this experiment is to reveal the mechanism of propagation and the hepatotoxicity in males.In this study, adult male frogs Rana chensinensis were exposed to different concentrations of10-7,10-6,10-5and10-4mol·L-1DEHP consecutively for20,30and40d respectively, then liver tissue was got from R.chensinensis at certain days. Hepatic cells are detected the expression of protein of Bax, Bcl-2, ER and Vtg by immunohistochemical technique, and apoptosis cells are tested by TUNEL. The results and the conclusions are listed as follows:1. The results of immunohistochemical technique of VTG show that VTG express in every treated group.The positive effect of VTG is yellow or brown in the cytoplasm.The results of the relative intensity of VTG show that the expression of VTG increased significantly in hepatic cells of R. chensinensis in DEHP treated group compared with the control group.In DEHP treated group, the expression of VTG is increased with the increasing concentration of DEHP, but not with the treated time.2. The positive effect of ER is yellow or brown in the cytoplasm and nuclear.The results of the relative intensity of ER show that the expression of ER increased significantly in hepatic cells of R. chensinensis in DEHP treated group compared with the control group.in DEHP treated group, the expression of ER is increased with the increasing concentration of DEHP. For the same concentration of DEHP treatment, the expression of ER did not significantly fluctuate with prolonged treatment. The change trend of ER is similar to the VTG. These results suggested that DEHP could caused the synthesis of VTG by dint of induce overexpression of ER in hepatocytes.3. The some of hepatic cells present deep black were detected by TUNEL technique. There are apoptotic cells in control group, AI is about3.43%. AI is20.37±2.13%in10-6mol·l-1DEHP treated for40d. AI is28.38±1.03%in10-4mol·L-1DEHP treated for40d, which is the highest. There exist dose-effect and time-effect relationship between DEHP and AI. Whereas, in10-5mol·L-1DEHP treated for40d, the AI is lowerer than treated for30d.4. Detecting the expression of Bax and Bcl-2by immunohistochemical technique in hepatic cells of R.chensinensis. The results show that the expression of Bax and Bcl-2are increasing with concentration along with the time. The expression of Bax is increasing with time, but decreasing with Bcl-2. Analyzing the linear regression relation between AI and the ratio of Bax/Bcl-2expression, the results show that significant positive correlation exist between AI and the ratio of Bax/Bcl-2expression in hepatic cell. It suggests that DEHP can increase the expression of Bax and decrease the expression of Bcl-2to induce the cell apoptosis.