Effects Of Cadmium On Apoptosis Mechanism Of Cock Sertoli-Germ Cell In Vitro Culture

Cadmium, a heavy metal of potential toxicity is extensively used in the industry and become one of the most important pollutants in the enviroment. A large amount of researches have indicated that Cadmium can lead to the functional decrease of reproduction in male animals and human and induce the nonreversible change such as apoptosis and necrosis of germ cells. Most of the studies focus on the rodent and human being, while only a few focus on the avian. This study took cock primary sertoli-germ cell as a model and added different concentration of CdCl2 into the culture medium. Mechanisms in cadmium-induced apoptosis of cock sertoli-germ cell were revealed through detecting the activation of the sertoli-germ cell, the capability of adherence, apoptosis, anti-oxidative function, DNA dammages, [Ca²⁺]i changes and gene expression. The results showed as follows:1. Determine the activation of cock sertoli-germ cell and the level of LDH by the tapan blue method ;MTT assay and dinitro-phenylhydrazine(DPNH) chromometry. The capability of adherence was observed. The result revealed that the activation of cock sertoli-germ cell cultured in vitro can be inhibited by cadmium and the capability of adherence as well as the sticky interaction between the spermatogenic cells and the support cells was influenced by Cadmium. IC50 of cadmium was 28.41±0.42μmol/L in 24h. Our result showed that cadmium had toxic effect to chock sertoli-germ cells.2.The apoptosis and necrosis percentage of the sertoli-germ cells were detected by AO/EB double fluorescence staining method. The results proved that low concentration cadmium can predominantly induce apoptosis, while high dose cadmium increased apoptosis and necrosis simultaneously.3.The effects of cadmium on DNA damage were detected by SCGE. The results showed that cadmium caused DNA damage, which exhibited dose-dependent effect. It revealed that the effects of cadmium on DNA damage of germ cells may be one of the mechanisms of damage of germ cells induced by cadmium.4.The result of detection on anti-oxidative function dedicated that cadmium lead to the decease of SOD activity and GSH-Px activity, the incease of MDA content and the anti-oxidative function, which lead to the oxidative stress. Possibly, this was one of the mechanisms of apoptosis induced by cadmium.5.CaMmRNA expression and [Ca²⁺]i were separatly detected by semiquantitative PCR and...