Experimental Study On The Effect Of Chaihu Shugan Powder On Ovulation Induction In PCOS Rats

Objective:To establish polycystic ovary syndrome rat model using the Letrozole,being intervened by Chaihu Shugan Powder.By observing the ovarian tissue morphology of rats,counting the number of mature follicles and luteum,testing the serum levels of sex hormones and measuring the integral optical density(IOD)of GnRH in hypothalamus,the research aims to clarify the intervention role and study the effective mechanism of Chaihu Shugan Powder in promoting the growth of follicle and ovulation due to further enrich the theoretical basis of treating polycystic ovary syndrome by soothing the Liver,as well as provid peri mental evidence for the treatment of PCOS ovulation disorders from the liver.Methods:The subjects were 32 finale Sprague-Dawley rats of SPF grade in age 6 weeks,which randomly and averagely divided into 2 groups,such as control group(n=12)and model group(n=20).The polycystic ovary syndrome animal mode is induced by Letrozole.After the modeling,the remaining rats were randomly divided into the Chaihu Shugan Powder group and the model control group(n=8)according to body weight,Chinese herbal grou was treated with Chaihu Shugan Powder,and the other two groups were filled with disinfect drinking water,1 times a day for 28 days.24 hours later after the last administration,rats were intraperitoneally injected 10%Chloral hydrate after weighing body weight.Then we opened the abdominal cavity,obtaine blood samples from abdominal aorta and cut out all ovarian tissues and hypothalamus.the sex hormone LH,FSH,T,and E2 in serum were tested by ELISA.The wet weight of bilateral ovariesin rats was weighed,and the organ coefficient of ovaries was calculated.the ovarian tissue morphology of rats were observed by hematoxylin eosin staining and then the number of mature follicles and corpus luteum were calculated.The expression level ofthe GnRH in hypothalamus were detected by Immunohistochemistry.Results:1 The comparison of control group and the successed animal model group:21 days after the modeling,the model group rested on anestrus for long time;Compared with the control group,rats in the model group gain weight markedly(P<0.05);H-E staining:in the model group,the follicles showed polycystic degeneration,no mature follicles,no oocytes and cumulus,and granular layer reduced to 1～3 layers.While follicles in different stages of development could be seen in the control group,also there were many granule cell layers,mostly were 8～9 layers.2 The difference of each group after treatment of drug intervention:(1)Body weight and organ coefficient of ovay:After 4 weeks of drug intervention,Chaihu Shugan Powder group resulted in a lower body weight and organ coefficient than those of the model control group,the difference was statistically significant(P<0.05).(2)The number of mature follicle and corpus luteum:H-E:the pathologic alterations of the ovary of Chaihu Shugan Powder group were significantly improved.The number of mature follicle and corpus luteum increased significantly(P<0.05).(3)Comparison of LH,FSH,T and E2 levels in serum:the T and LH in the model group were significantly higher than that in the normal group(P<0.05),and the FSH was lower than that of the normal group(P<0.05).The levels of LH and T in the Chaihu Shugan Powder group decreased(P<0.05)and the FSH level increased,but it was still lower than that of the normal group,the difference was statistically significant(P<0.05).The level of E2 were similar in each group,and there was no difference(P>0.05).(4)The results of immunohistochemistry:compared with the normal group,the IOD of the model group increased significantly(P<0.01).Compared with the model group,the IOD of the group of Chaihu Shugan Powder group was lower than that in the model group,and the difference was statistically significant(P<0.05).Conclusions:1 Chaihu Shugan powder has the effect of inducing ovulation in polycystic ovary syndrome rats.2 The promoting ovulation effect of Chaihu Shugan San is probably achieved by down regulating the expression level of GnRH in hypothalamus and improving the endocrine function of hypothalamic-pituitary-ovarianaxis.