| Objective To investigate the expression of BMP-7 in ovarian of POF rats and the mechanism of Zuogui pill in preventing and treating ovarian function decline,and to provide theoretical basis for clinical application of Zuogui pill.Methods A total of 60 8-week-old SD rats with normal estrus cycle were screened and randomly divided into 6 groups:blank group,model group,bujiale group and Zuogui pill（high,medium and low）dose groups.The rat model of premature ovarian failure was established by intraperitoneal injection of cisplatin（6mg/kg）.The changes of external performance of rats in each group were observed daily.During the modeling period,vaginal exclined cells were continuously smeared to observe the estrous cycle changes of rats.After modeling,except for blank group and model group,the other groups were given different doses of Zuogui Wan and Bujiale for continuous intragastric administration for 4 weeks.After sampling,the contents of FSH,LH and E2in serum were determined by enzyme-linked immunosorbent assay.The changes of ovarian tissue morphology were observed by HE staining.The expression of BMP-7 and Cx37 in ovarian tissue was detected by immunohistochemistry.The experimental data were analyzed and processed by SPSS 19.0software.Results 1.General situation and changes of estrus cycle:compared with blank group,after modeling,the rats showed depression of spirit,dark hair color,less food and water intake,weight loss,and obvious disorder of estrus cycle in the rats with vaginal exhumed cells and smear removal;After drug intervention,the hair color of the rats gradually recovered luster,the mental state improved,the amount of food and drinking water gradually returned to normal,and the weight of the rats increased significantly.2.Changes of serum sex hormone levels:Compared with the blank group,the serum sex hormone levels of rats were in a state of disorder after modeling,and the contents of serum FSH and LH increased significantly.The content of serum E2decreased significantly.After drug treatment,the contents of FSH and LH in serum of rats decreased in different degrees compared with model group,and the contents of E2increased in different degrees（P<0.01）,and the effect of Bujiale group and Zuogui pill high-dose group was similar（P>0.05）.3.The changes of ovarian histomorphology:compared with the blank group,the ovarian tissue structure of the model group was unclear,the cortex was thinned,the number of follicles in each development stage was reduced,only primitive follicles were found,and the interstitial necrosis was obvious.After drug intervention,the ovarian tissue structure of rats in the treatment group was significantly improved compared with that in the model group,the number of follicles in each development stage was increased,and the hierarchical arrangement of granulosa cells was changed.4.Expression of BMP-7 and Cx37 protein in ovary of POF rats:BMP-7 was mainly expressed in oocytes.Compared with the blank group,the protein expressions of BMP-7 and CX37 in the ovaries of rats in the model group were significantly decreased,the differences were statistically significant（P<0.01）.After drug intervention,BMP-7 and Cx37 in all treatment groups were increased to varying degrees,and the changes were most obvious in Bujiole group and Zuogui pill high-dose group.Compared with model group,the difference was statistically significant（P<0.01）.Conclusion1.Zuogui pill can effectively regulate the general condition and serum hormone level of premature aging rats,reduce the contents of FSH and LH,increase the content of E2,and play a role in improving ovarian function.2.BMP-7 protein is mainly expressed in rat ovary oocytes.After cisplatin induced premature ovarian failure,the expression of BMP-7 protein in rat ovary is significantly reduced.3.Zuogui pill can up-regulate the expression of BMP-7 and Cx37 proteins in the ovary of premature aging rats,which may be achieved by promoting the early development of primitive follicles and improving the gap junction function. |
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