Experimental Study On The Effect Of Tonifying Kidney And Resolving Phlegm On Follicular Development In The Obese PCOS Rats

Objectives:In this study,Letrozole and high-fat were used to induced the model of obese polycystic ovary in rats,and the effects of tonifying kidney and resolving phlegm?TKRP?method on serum sex hormones,ovarian morphology and expression of Smad3,Cyp19al and CCND2mRNA in granulosa cells of ovarian tissue in obese PCOS female rats were observed.Investigate the mechanism of TKRP method in regulating follicular development of PCOS,so as to provide a scientific basis for the treatment of obese PCOS follicular developmental disorders.Methods:The 7-to 8-week-old SD female rats were selected to prepare obese PCOS model by letrozole and high-fat oral gavage.According to the rat vaginal exfoliated cytology smear and changes of body weight,successful models were selected.According to the experimental requirements,the rats were divided into model group,high dose group of traditional Chinese medicine,middle dose group of traditional Chinesemedicine,low dose group of traditional Chinese medicine,Diane-35 group,and blank control group.Twenty-one days after intervention with the TKRP herbs,the changes of serum FSH,E₂,LH and T were detected by radioimmunoassay,the pathological changes of ovarian tissue were observed by using hematoxylin-eosin staining,the mRNA expressions of Smad3,Cyp19al,and CCND2 in ovarian granulosa cells were measured by Real-time PCR.Immunohistochemical method was used to detect the expression of Ki-67 in granulosa cells of rat ovaries.Results:?1?Compared with the model control group,the body mass growth rates of the high-,middle-dose group of traditional Chinese medicine and the Diane-35 group decreased,the difference was statistically significant?P<0.05?;?2?Compared with the model group,the number of primary follicles,sinusoidal follicles,and cystic follicles in the traditional Chinese medicine and Diane-35 group decreased,and the number of mature follicles and corpus luteum increased,and the difference was statistically significant?P<0.05?;?3?Compared with the model group,serum FSH,E₂ in high-,middle-dose group of traditional Chinese medicine and Diane-35 group increased,and the difference was statistically significant?P<0.05?.The T content was decreased in high-,middle-dose group of traditional Chinese medicine and western medicine group.The decrease of western medicine group was obvious,and the LH content in the middle dose group of traditional Chinese medicine and Diane-35 group were decreased.The difference was statistically significant?P<0.05?;?4?Compared with the blank control group,the expression of Smad3,Cyp19al and CCND2 mRNA in the ovarian granulosa cells of the model groups decreased?P<0.05?;Compared with the model groups,the expression of Smad3,Cyp19al,and CCND2 mRNA in ovarian granulosa cells was increased in the high and middle dose groups?P<0.05?.?5?Compared with the blank control group,the expression of Ki-67 in the granulosa cells of the ovarian tissue of the model groups decreased?P<0.05?;Compared with the model groups,the expression of Ki-67 in the granulosa cells of the ovarian tissue of the middle dose group of the Chinese medicine and the positive control groups elevated?P<0.05?.Conclusions:?1?TKRP method can suppress polycystic ovaries of obese PCOS rats and improve follicular developmental disorder.?2?TKRP method can correct the state of sex hormone disorders in the body and regulate ovarian function by reducing the levels of T and LH and increasing the levels of FSH and E₂.?3?TKRP method is possibly via promoting the TGF?/Smad3 signal transduction of ovary granulosa cells,thereby enhancing the expression of Cyp19al mRNA and promoting the conversion of androgens into estrogens to maintain the body's sex hormone homeostasis;and it maybe through regulating the TGF?/Smad3 signal transduction of ovary granulosa cells,thereby enhancing the expression of CCND2,promoting granulosa cell proliferation and follicular development and maturation.