| Objectives 1 To investigate the effect of Bushen Tiaochong Recipe induced ovarian reserve senescence gene P53,ovarian tissue dysfunction rat gonadotropin releasing hormone receptor GnRHR on cyclophosphamide;2 To observe the expression of P53 and GnRHR in poor ovarian reserve function in ovary of rats,and provide a theoretical basis and ideas for TCM clinical treatment of ovarian reserve low function.Methods 12 week old clean female SD rats were randomly divided into the blank group,the model group,the model group,the group of tonifying kidney and punching square high,middle and low dosage groups,after making the model group,the high,middle and low dose group of tonifying kidney and punching prescription,western medicine group,10 rats in each group.Except for the normal feeding of the blank group,the rats in the rest of the rats were injected with cyclophosphamide 1 times daily at the dose of 75mg/kg,and the rats were prepared for 1 days?the end of the 14 d model?to prepare the rat ovarian reserve function model.At the same time,the model group was given the dosage of 24,12 and6g/kg in the dose of Tonifying the kidney and punching the stomach to prevent the stomach.After the model,the group was given the dosage of 24,12,6g/kg of the tonifying kidney and punching prescription after the model 14 days,and the two groups were given36 d after continuous irrigation,and the western medicine group was given the combination of estrogen 0.075mg/kg to estrogen.Gavage was given with the method of gushing kidney and regulating Chong decoction.The data obtained from the test are all analyzed by SPSS21.0 statistical tools.The data obtained are represented by mean+standard deviation????±s?.The changes of general behavior and estrus cycle were observed in rats.The levels of serum follicular stimulating hormone?FSH?,luteinizing hormone?LH?,serum estradiol?E2?and anti Muller hormone?AMH?were detected by enzyme linked immunosorbent assay?Elisa?.Immunohistochemistry and real-time fluorescein quantitative PCR method were used to detect the aging of ovarian tissue in rats Due to the protein and gene content of P53 and gonadotropin releasing hormone receptor GnRHR,the effect of tonifying kidney and punching Recipe on P53 and GnRHR in rats with cyclophosphamide induced hypoplastic ovarian reserve was discussed.Results 1 The effect on the general condition of the rats:the rats in the blank group had good function in every aspect.The model group began to be depilatory,poor spirit,the number of urination and the decrease of the number of stools from the fourth days of model building.A slight improvement,the loss of hair loss,the first decrease in weight,and the first decrease in weight,and the most obvious recovery in the high dose group of the tonifying kidney and punching prescription,and the expression of the model group in the early stage of the model group.After the drug was given to the stomach,the body weight first descended and then gradually showed a stable state,the mental activity,a small amount of hair removal,drinking water,and stool.The highest level of recovery was in the high dose group.2 Influence on the estrous cycle of rats:the estrous cycle in the blank group was in the normal range,about 45d during the whole experiment.The estrous cycle of the model group extended to 710d,even the disorder,the epithelial cells in the vaginal smear were more in the estrus interval,and the estrous cycle of each group was compared with the blank group.The difference was statistically significant?P<0.05?.Compared with the western medicine group,the difference in the estrous cycle of the kidney tonifying and punching prescription group was statistically significant?P<0.05?,and the difference between the middle and low dose groups of the kidney tonifying and punching prescription after the model group was statistically significant?P<0.05?.3 The effect of serum sex hormones in rats:compared with the model group,the serum levels of FSH,LH,AMH and E2 in each group were statistically significant?P<0.01?.Compared with the blank group,the serum level of FSH and LH in the model group was significantly higher,the level of serum AMH and E2 decreased significantly?P<0.05?,and the low dose of tonifying kidney and punching prescription in the model group was low.The serum levels of FSH,E2 and AMH were statistically significant?P<0.05?.There was significant difference in serum E2 and AMH levels in the middle dose group of the invigorating group after making the model group?P<0.05?,and the difference of serum in the low dose group was statistically significant?P<0.05?,and the difference of serum E2 and AMH levels in the group of invigorating kidney and tonifying the kidney in the low dose group was compared with the western medicine group.There was statistical significance?P<0.05?;after modeling,the serum level of the low dose group was statistically significant?P<0.05?.4 The effect of the ovaries on the ovaries of rats:the ovarian structure of the rats in the blank group was clear.The follicles of the beginning,the preantral,sinus and atresia follicles were seen at all levels.The follicles developed well and the volume was larger.The follicles at all levels in the model group were less,the follicle developed and atrophied,the volume was small,and the number of the granules in the follicle was few.Compared with the blank group,there was no significant difference in the development of follicular development in each group.Compared with the model group,the quality of follicular development in each group was better.Compared with the western medicine group,the follicular development was poor and the volume slightly atrophied,and the quality of the follicular development in the middle and low dose group after the model group was slightly worse.5 The effect on the expression of P53,GnRHR protein and mRNA in the rat ovary:compared with the model group,the expression of P53 protein and mRNA decreased obviously,the expression of GnRHR protein and mRNA increased obviously,the difference was statistically significant?P<0.01?.Compared with the blank group,the expression of P53 protein and mRNA in the ovary increased obviously,GnRHR in the ovary.GnRHR The expression of protein and mRNA was significantly decreased?P<0.05?,and the difference of P53,GnRHR protein and mRNA expression in the low dose group of tonifying kidney and Chong Chong recipe group was statistically significant?P<0.05?.Compared with the western medicine group,the difference of P53,GnRHR egg white and mRNA expression in the low dose group of tonifying kidney and Chong Chong recipe group was statistically significant?P<0.05?.There was a significant difference in the expression of P53 protein and mRNA between the rats in the middle dose group of the Bushen tune and the Chong Decoction group?P<0.05?.Conclusions 1 Cyclophosphamide 75mg/kg intraperitoneal injection 1 times a day,a total of 1 days,can lead to rats' estrous cycle prolonged or disorder,the wet weight and body mass of the ovary and uterus are reduced,the serum level is low female high,the rat ovarian reserve function is successfully prepared.2 The recipe for Bushen and Chong Chong can effectively prevent and treat the low ovarian reserve function,in which the prevention and treatment effect of the high dose group of tonifying kidney and punching prescription is the best,which is equivalent to the combination of estrogen group.3Chinese medicine can better protect the ovary,and significantly reduce the damage of antitumor drugs to the reproductive system. |
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