| Objectives By observing the Bushen Tiaochong Prescription on expression of Changes in hormone levels, the change of the ultrastructure of the uterus and ovary and changes of ovarian, uterine related apoptosis and immune factors of Female rats which were because of cyclophosphamide the DOR. Explorer the prevention?improvement and adjustment effects of Bushen Tiao Chong Prescription on DOR, as well as the prevention of drug damage, to provide innovative ideas and measures for the prevention and treatment of DOR and DOR in Traditional Chinese Medicine.Methods 12 week old SD level SPF female rats were randomly divided into Normal control group, Model group, Bushen Tiaochong prescription high?middle?low dosage prevention group, estrogen prevention group, Bushen Tiaochong prescription high ?middle?low dosage simultaneous medication group, estrogen simultaneous medication group, Bushen Tiaochong prescription high ?middle?low dosage cure group, estrogen cure group. In addition to the control group, the other groups were given intraperitoneal injection 75mg/kg cyclophosphamide to prepared once model, Before making the model with Bushen Tiao Chong Prescription of high, medium or low dose or the combination of 0.075mg/kg estrogen to intervene. After and meanwhile the model was made, the Bushen Tiao Chong Prescription was used to adjust the high, medium or low dose or the combination of estrogen for treatment. To observe the behavioral changes of rats, estrous cycle, the change of the ultrastructure of the uterus and Ovary computing the index of uterus and Ovary. ELISA method was used to detect serum FSH, LH, INHB, AMH in rat serum; RIA method was used to detect serum E2 in rat serum. Ovarian related apoptosis factor(Bax), Caspase-3 protein, m RNA protein, RT-PCR protein, Bcl-2, were detected by immunohistochemistry and semi quantitative method. Fluorescence quantitative PCR method was used to detect the content of TNF-? alpha, m RNA gamma IFN-? in rat ovaries.Results 1 Effects on the rat's emotional cycle: The cycle of rats oestrous in blank control group?Low dose Bushen Tiao Chong Prescription treatment group?Low dose Bushen Tiao Chong Prescription simultaneous medication groups?conjugated estrogens groups were 4-5 days; Sixty percent of model groups the cycle of rats oestrous were extended to the day of execution, Forty percent were 6-8 days; All of the high dose Bushen Tiao Chong Prescription groups were 5-8 days; All of the middle dose Bushen Tiao Chong Prescription groups were 4-6 days; Low dose Bushen Tiao Chong Prescription prevention group were 4-6 days. 2 Effect of sex hormone level in rats: The level of E2?INHB?AMH in model group is significantly lower than other groups, besides the difference was statistically significant(P<0.05); Each groups the level of E2?INHB?AMH were significantly increased by given Bushen Tiaochong prescription of traditional Chinese medicine and combined with estrogen. The level of FSH and LH in model group were increased, compared with the other groups, the difference was statistically significant(P<0.05); Each groups the level of FSH and LH were significantly increased by given Bushen Tiaochong prescription of traditional Chinese medicine and combined with estrogen. The model group compared with the other groups the ratio of FSH/LH was increased, and the ratio is greater than 2, Each groups the ratio of FSH/LH were significantly decreased by given Bushen Tiaochong prescription of traditional Chinese medicine and combined with estrogen. 3 The changes of rats uterus, ovary index: The model group compared with the other groups index of uterus, ovary index was decreased, give each groups the uterus indexof Bushen Tiaochong prescription of traditional Chinese medicine and combined with estrogen, the ovary index can be increased, the overall comparison of the prevention group level is close to the normal level. 4 The changes of expression level in rat ovary?apoptosis related factor of uterus: By immunohistochemical method in model group ovarian, uterine Bax, Caspase-3 protein expression was significantly higher than that in other groups(P<0.05), each groups the level of Bax?Caspase-3 were decreased by given Bushen Tiaochong prescription of traditional Chinese medicine. Ovary model group, the expression level of Bcl-2 protein in uterus was significantly lower than other groups(P<0.05). Each groups the level of oarium Bax?Bcl-2?Caspase-3 m RNA were identical with the result of immunohistochemical. The overall comparison of prevention group was better than that of the same drug group, simultaneous administration group was better than the treatment grou. 5 The changes of expression level in rat related cell factor TNF-??IFN-? m RNA: The model group compared with the other groups the expression level TNF-??IFN-? m RNA in rats oarium was increased, besides the difference was statistically significant(P<0.05). each groups expression level of TNF-??IFN-? m RNA were decreased by given Bushen Tiaochong prescription.Conclusions 1 Cyclophosphamide 75mg/kg intraperitoneal single injection can cause damage to the ovary function in rats, prolong the emotional cycle, success rat ovarian reserve function decline model preparation. 2 The kidney nourishing prescription can regulate the endocrine level of DOR rats through the regulation of hypothalamus pituitary ovary axis. Improving ovarian reserve capacity in rats, the prevention group can protect the ovary and reduce the damage of the ovary before the ovary is damaged, reduce the damage to the ovaries. The mechanism of action is inferred to be the kidney regulating prescription to damage the ovarian antigen target cells by inhibiting the apoptosis factor. Baffling the development of the cell death, slowing down the rate of follicular atresia, promotes the ovarian tissue repair, restore ovarian function as a whole. 3 Bushen Tiao Chong Prescription as DOR treatment efficacy from high to low were low dose group, middle dose group, high dose group. The effect of low dose group and estrogen group was equivalent. 4 The prevention group was superior to the group in the same time. Simultaneous administration group was better than the treatment group. Treatment of tumor and other diseases with cyclophosphamide in clinical, can be given to the Bushen Tiao Chong Prescription to prevent, reducing the damage of chemotherapy drugs to the gonad. |
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