Comparative Study Of The Effect About Bushen Treatment And Shugan Treatment On Anovulatory Model Rats

Objective: Through the study of comparative Bushen method and Shugan method anovulation similarities and different of the impact of rats neurotransmitter and hormone-secreting pituitary adenoma explore the mechanism of the Bushen and Shugan treatment to ovulation disorder mechanism in order to in enrich reproductive theory in traditional Chinese medicine, promoting the development of Chinese Gynecology.Method: 204 healthy female Wistar rats of 3 months old ,weighting 180-250g were fed as healthy,randomly 170 of these rates were improved as the method of Zheng Jia run,Liang Wen bo inc. Tripterygium 30mg/kg/d (doses of medication for clinical adult 20 times of the amount of times), by intragastric administration, 57 days in a row.weighting every week,adjusting the delivery quantity). Modules of vaginal epithelial cell smears two consecutive cycles (a cycle of 4-5 day) estrous cycles become irregular, mainly for the cycle is not complete and cycle extended,suggesting that anovulatory rat model induced by tripterygium successfully. Intragastric administration of distilled water to the remaining 34 as a normal control group.204 healthy female SD rats of 9-day age from the same batch, weighing about about 15g, were adopted in our research. Randomly 170 of those rats were made as models by subcutaneous injection of testosterone propionate at once 1.25mg/(0.05ml/)at nucha. All these 204 rats were weaned when they were 22-day old, eating free, same conditions feeding. When they were 70-day old, their vaginals debouched. We began to make the smear of the vaginal epithelial cell smears two consecutive cycles (a cycle of 4-5) for the model group, continuous angle of vaginal epithelial cells suggesting anovulatory rat caused by androgen successfully. The remaining 34, injected neutral Camellia oil 0.05ml as a normal control group ,following the methods of Yu Jin etc.The two batchs of model rats were randomly divided into five groups: the model group, the Bushen Tiaojing Recipe high-dose group, the Bushen Tiaojing Recipe low-dose group, the Xiaoyao pill high-dose group, the Xiaoyao pill low-dose group, each group was made uP of 34 rats; the treatment groups were drenched with Bushen Tiaojing Recipe and Xiaoyao pill, model groups and normal groups were drenched with distilled water once a day. Treatments continued for 5 weeks, and made the smear of vaginal epithelial cells for 2 successive sexual cycle. Prohibited feeding them within 8 hours after administration, each group were decaPitated at proestrus and estrus periods, centrifugation to prepare for testing. Aseptic in the ice tray to remove the hypothalamus. The scope of the hypothalamus : rostral to the anterior, caudal to posterior of the mammillary bodies,the lateral hypothalamus to the lateral sulcus, dorsal to the level of anterior.The tissues includes the bottom side of hypothalamic, anterior hypothalamus and preoptic area. Then weight the tissue. Homogenate the left side of the hypothalamus adding 1 mL 1 mol/L HAC, 4℃4 000 r / min centrifugal 20 minutes, take out of the supernatantadd and add1ml NaOH(1 mol/L) to counteract it. Conserve them by liquid nitrogen inorder to detection. Sonserve the right side of the hypothalamus in the liquid nitrogen for later detection. Measuringβ-EP by radioimmunoassay and NE,DA,5-HT by fluoroimmunoassay.Results:1 Modeling results:1.1 Model Rat of Triptolide -induced ovulation disorders Compared with the normal group,β-EP,5-HT of the model group are both significantly increased(P<0.05,P<0.01),DA,NE are both significantly decreased(P<0.01,P<0.01)1.2 Model Rat of Androgen -induced ovulation disorders normal group,β-EP,5-HT of the model group are both significantly increased(P<0.01 , P<0.01),DA,NE are both significantly decreased(P<0.05,P<0.05)。 2 The impact of Bushen Tiaojing Recipe to neurotransmitter of the two kinds of ovulation disorder model rat2.1 Anovulatory rats induced by tripterygium fed with Bushen Tiaojing Recipe, compared with model group:①low dose in proestrusβ-EP is significantly decreased(P<0.05), but not to the normal levels(P<0.05); high-dose group in proestrus and post-estrous are both significantly decreased(both P<0.01)and to the normal levels(both P>0.05)②low dose and high-dose group in proestrus and post-estrous NE are both significantly increased(both P<0.01), and to the normal levels(both P>0.05)③low dose group in proestrus and post-estrous DA is not to the normal levels(both P<0.05), high-dose group in proestrus DA is significantly increased(P<0.01)and to the normal levels(P>0.05), high-dose group in post-estrous is not to the normal levels(both P<0.01).④low dose group in proestrus and post-estrous 5-HT are both significantly decreased(both P<0.01), and to the normal levels(P>0.05, P>0.01); high-dose group in proestrus and post-estrous are not to the normal levels(both P>0.05) (both P<0.01).2.2 Anovulatory rats induced by androgen fed with Bushen Tiaojing Recipe, compared with model group:①l ow dose in proestrusβ-EP is not to the normal levels(P<0.01); high-dose group in proestrus is significantly decreased(P<0.05)and to the normal levels(P>0.05)②h igh-dose group in proestrus NE is significantly increased(P<0.01), and to the normal levels(P>0.05) ,low dose group in proestrus n proestrus NE is significantly increased(P<0.05), but not to the normal levels(P<0.05)③low dose group in proestrus and post-estrous DA is not to the normal levels(bothP<0.05), high-dose group in proestrus DA is not to the normal levels(P<0.05), high-dose group in post-estrous significantly increased(P<0.05), and to the normal levels(P>0.05,);④low dose and high-dose group in proestrus and post-estrous 5-HT are both significantly decreased(P<0.01,P<0.05,both P<0.01), and to the normal levels(P>0.05, P>0.01, both P>0.05). 3 The impact of Xiaoyao pill to neurotransmitter of the two kinds of ovulation disorder model rat3.1 Anovulatory rats induced by tripterygium fed with Xiaoyao pill, compared with model group:①l ow dose in proestrus and post-estrusβ-EP is significantly decreased(P<0.01), but not to the normal levels(P<0.05); high-dose group in proestrus is significantly decreased(P<0.01)and to the normal levels(P>0.05)②l ow dose and high-dose group in proestrus and post-estrous NE are both significantly increased(bothP<0.01), and to the normal levels(bothP>0.05)③low dose group in proestrus and post-estrous DA is not to the normal levels(P < 0.01,P < 0.05), high-dose group in proestrus DA is significantly increased(P<0.01)and to the normal levels(P>0.05).④Low dose and high-dose group in proestrus and Post-estrous 5-HT are both significantly decreased(both P<0.01), and to the normal levels(both P>0.05).3.2 Anovulatory rats induced by androgen fed with Xiaoyao pill, compared with model group:①l ow dose in proestrusβ-EP isignificantly decreased (P<0.01),but not to the normal levels(P<0.05); high-dose group in proestrus and post-estrous are both significantly decreased(P<0.01,P<0.05)and to the normal levels(both P>0.05)②low dose in proestrus and post-estrous NE are both significantly increased(P<0.01,P<0.05), and to the normal levels(bothP>0.05)③low dose and high-dose group in proestrus and post-estrous DA is not to the normal levels(both P<0.05)), .④low dose and high-dose group in proestrus and post-estrous NE are both significantly decreased(P<0.01,P<0.05,both P<0.01), and to the normal levels(both P>0.05).4 The differences between the effect of Bushen Tiaojing Recipe and Xiaoyao pill to eurotransmitter of the two kinds of ovulation disorder model rat 4.1 Triptolide induced ovulation disorder model group Low dose of Bushen Tiaojing Recipe and Low dose of Xiaoyao pill in proestrusβ-EP have significant difference (P<0.05), high-dose of Bushen Tiaojing Recipe and high-dose of Xiaoyao pill in Post-estrousβ-EP have significant difference (P <0.05). High-dose of Bushen Tiaojing Recipe and high-dose of Xiaoyao pill in proestrus NE have significant difference (P < 0.05). Low dose and high-dose of Xiaoyao pill in Proestrus and post-estrous 5-HT have significant difference with high-dose of Bushen Tiaojing Recipe(P<0.05, P<0.01;均P<0.05). High-dose of Bushen Tiaojing Recipe and high-dose of Xiaoyao pill in Proestrus DA have significant difference (P<0.01),high-dose of Xiaoyao pill and high-dose of Bushen Tiaojing Recipe in Post-estrous DA have significant difference (P<0.05).4.2 Rogen induced ovulation disorder model group Low dose of Xiaoyao pill and Low dose of Bushen Tiaojing Recipe in proestrusβ-EP have significant difference (P<0.01). High-dose of Bushen Tiaojing Recipe and high-dose of Xiaoyao pill in proestrus NE have significant difference (P<0.05). Low dose a of Xiaoyao pill in post-estrous and Low dose of Bushen in post-estrou NE have significant difference (P<0.05). High-dose of Bushen Tiaojing Recipe and high-dose of Xiaoyao pill in Post-estrous have significant difference (P<0.05).Conclusions:(1) Neurotransmitter in the hypothalamus of the two kinds of ovulation disorder model rat both happens disorder.(2 )Bushen Tiaojing Recipe and Xiaoyao pill by regulating the levels of neurotransmitters, play a role in regulating gonadal axis,and treatment the ovulation disorders. Bushen Tiaojing Recipe and Xiaoyao pill have differents in regulating of hypothalamic neurotransmitters .