Research On Chemotherapy-induced The Change Of Activety And Expression Of Cytochrome P450 Aromatase In Mice Granulose Cells And Intervention Effect Of Siwuheji

BackgroundWith the widespread use of adjuvant chemotherapy in early breast cancer patients, the long-term effects of treatment are becoming increasingly important. Chemotherapy-induced amenorrhea, with resulting infertility and prolonged exposure to risks and symptoms of menopause, has bin receiving increasing attention in recent years.ObjectiveTo investigate the change of activity and expression of cytochrome P450 aromatase in mice granulose cells by cisplatin-induced, and its relation with the Smad sign transduction pathway; to explore intervention effect of Siwuheji and its mechanism, then to provide basis for clinical therapy. MethodBased on the animal model of chemotherapy-induced POF and primary culture of granulose cell from the ovaries of rats, by using the techniques of serological method, radio-immunologic, immumohistochemistry and electron microscope, study the mechanism of intervention effect of Siwuheji.Result1. Studay on the feasibility of chemotherapy-induced POF therapy with SiwuhejiICR mice were i.p. injected daily with cisplatin at doses of 3.0 mg/Kg body weigh daily for 7 days, and were supplemented Siwuheji for 14 days. Compared with the control group, the biological behavior character were changed, the level of GSH and the activity of GSH-Px, SOD and complex I were decreased, the level of MDA and the activity complex II were increased, the levels of RBC and HGB were increased on the 4th day, the levels of LY and PLT were increased on the 7th day. Those changes could be relieved by supplemented Siwuheji.2. Protective effects of SiWuHeJi on cisplatin induced ovarian dysfunction in miceICR mice were i.p. injected daily with cisplatin at doses of 3.0 mg/Kg body weigh daily for 7 days, and were supplemented Siwuheji for 14 days. Compared with the control group, serum levels of ovarian hormones were decreased and serum levels of FSH/LH radio were increased. Development of preantral follicle stopped, with the number of atretic follicle increased and local hyperplasia of stroma which have change of fribrosis. Those changes could be relieved by supplemented Siwuheji.3. Found a means for judgement the result of counting granulose cell cultured with CDDPUsing tow cell count, MTT and cck-8, contrasting the change of absorbance and absorbance radio at three time stage, it was made clear that MTT and CCK-8 were combined together was a precise means.4. Effects of Siwuheji on proliferation and secretory function of granulose cell cultured with CDDPWhile the granulose cells were cultured with CDDP, the numerical value of counting was lower and the levels of E2 and P were decreased. Those changes could be relieved by doped pharmacoserum of Siwuheji.5. Effects of Siwuheji on expression of aromatase and relative signal modulinExpression of aromatase and signal modulin of granulose cell in ICR mice injected with CDDP was decreased. The levels of expression could be increased by supplemented Siwuheji.ConclutionBased on the animal model and primary culture of granulose cell from the ovaries of rats, it was made clear that expression of aromatase and signal modulin of granulose cell was decreased by cisplatin-induced. It could be relived by doped Siwuheji. Furthermore, other changes induced by CDDP also could be relived. It is possible believed that Siwuheji could be useful for chemotherapy-induced POF.