The Study On Cadium Induced Apoptosis Of Mouse Granulosa Cells And Correlation Influence Factor

Cadmium is a contamination of environment and industry. It has obvious toxicityfor the reproductive of female mammalian, which could cause the ovary tissuepathological change, disturbed ovulation and fecundation, lead to sterility provisional,both of the accepters of estrogen and progesterone were affected by cadmium, and theexpression of genes as well.Reserch Aim: The effect of cadmium on the ovarian granulose cells wasdefecated and the mechanism was clarified. The theory of cadmium-induced thetoxicity on gonad was renewed. The theoretic evidence was provided for later study.Research methods:Prepubertal female mice as object injected with severaldose cadmium chloride after subcutaneous injected with PMSG.The follicular atresiawas detected by pathological method;the apoptosis of granulosa cells were detectedby TUNEL; the content of MDA and the activeness of SOD were detected byconventional method; the p53 protein expression was detected by immunityhistochemistry and the semi-quantitative RT-PCR.Research Results: The TUNEL result showed that the apoptosis percent ofgranulose cells in high, midst and low-dose groups are 14.524%, 11.5764%, 9.8762%respectively after 48 hours, which are higher than control groupsignificantly(0.5426%, P＜0.01). The apoptosis percent in control, high, midst andlow-dose groups grow up to 10.2064%, 21.4146%, 15.8864%, 12.4721% respectivelyafter 96 hours. The result showed that the apoptosis in diverse degree were occurredin every groups, the apoptosis index increase with dosage rising, there is extremesignificant difference (P＜0.01). The percent of atresic follicular increased withdosage rising (P＜0.05). To examine the content of MDA and the activeness of SODby conventional method, the content of MDA increased with dosage rising, there isextreme significant difference (P＜0.01) between testing group and control group inthe two time section of 48h and 96h. The activeness of SOD decreased with dosagerising, there is significant difference between control group and testing group on 48 hours, there are extreme significant difference (P＜0.01) between control group andtesting group except dosage of l mg/kg (P＜0.05) on 96 hours. The result ofexpression of immunity histochemistry and the semi-quantitative RT-PCR examiningp53 protein expression showed that p53 express significant difference between testinggroup and control group (P＜0.01) except dosage of 1mg/kg (p＜0.05) on 96 hoursnot by immunity histochemistry.Research Conclusion:1. Cadmium can accelerate the apoptosis of granulosecells and is related with dose. 2.Cadmium accelerate atresic follicular, and canincrease the constitution ratio of atresic follicular . 3. Cadmium can increase thecontent of MDA of granulose cells and can decrease the activeness of SOD. 4. Thep53 gene participated in the regulation of the apoptosis of granulose cells, cadmiumcan enhance the expression of p53 protein in granulose cells. 5. Apoptosis ofgranulose cells induced by cadmium is the result of free radical and regulative geneaffected together...