| Objective:This experiment based on pre study, to further explore to Bushen Huoxue legislation new regression kidney Pill on induced by Tripterygium Glycosides follicular developmental disorder model rats ovarian tissue related factors and molecular mechanism of the effects of protein, to evaluation method of tonifying kidney and activating blood on ovarian local environmental intervention effect, for clinical application of the method provides objective basis.Method:76 normal estrous cycle of female SD rats were selected and randomly selected 18 rats as normal control group given distilled water by gavage. The remaining 58 only according to the previous study, exploring the dose. Tripterygium Glycosides 75mg/(kg.30d) irrigation stomach replication follicular development deficits in animal models. After the end of the modeling with the machine from each of the eight experimental animals (16) to verify the modeling is successful. Successful model rats were randomly divided into 5 groups, namely, model blank group, Chinese medicine high dose group, low dose of Chinese medicine group, Ding Kundan group, western medicine group, each group of 10.Blank model group and normal control group were given distilled water by gavage; Chinese medicine groups give new regression kidney shot high, low dose treatment; western medicine group given acid estradiol acetate and methyl hydroxy progesterone sequential therapy; Dingkun Dan group given Dingkun Dan treatment; gastric perfusion during vaginal exfoliated cells of the examination.Blood samples were collected from the femoral artery after 2 weeks of continuous filling of stomach, and the content of serum FSH and E2 were detected;After blood collection, the rats were killed, open speed and the ovaries were removed and weighed, side immediately placed in 10% neutral formalin preserved, prepared for ovarian sections stained with hematoxylin and eosin (he) staining, the other side implanted with liquid nitrogen preservation were measured of IGF-1R, FOXO3a, HIF-la protein relative expression quantity assay (WB) and IGF-1R, FOXO3a, HIF-lamRNA relative expression was detected by polymerase chain reaction (PCR).Results:(1) Compared with the normal control group, the rats in the model group were prolonged, the weight change rate, the wet weight of ovary, the ovary index decreased, the number of growing follicles at all levels, the decrease of corpus corpus, the number of atresia follicles increased.(2) Compared with the normal control group, the serum FSH level of the model blank group was increased, the E2 content was decreased, and the difference was statistically significant (P<0.05).Compared with that of the blank model group, high dose of Chinese medicine group, Dingkun Dan FSH group content decreased significantly, E2 levels were significantly increased, the difference is statistically significant (P< 0.05), and the high dose of Chinese medicine group is the best.(3) Compared with the normal control group, the relative expression of FOXO3amRNA in the model blank group was increased, and the difference was statistically significant (P<0.05).With that of the blank model group comparison, the treatment group FOXO3amRNA relative expression amount decreased, with Chinese medicine high dose group decreased significantly, and the difference is statistically significant (P<0.05); other drug groups was no statistical significance (P>0.05).(4) Compared with the normal control group, the relative expression of IGF-1RmRNA in model blank group had a decreasing trend, but the difference was not statistically significant (P> 0.05).Compared with model blank group, the relative expression of IGF-1RmRNA in high dose group was higher than that in traditional Chinese medicine group, but the difference was not statistically significant (P>0.05).(5) There was no significant difference in the relative expression of HIF-lamRNA (P> 0.05) in the ovarian tissue of the rats in each group.(6) Compared with the normal control group, the relative expression of HIF-la protein and IGF-1R protein in the model blank group were decreased, and the relative expression of FOXO3a protein was increased, and the difference was statistically significant (P<0.05).With that of the blank model group compared each drug treatment group of IGF-1R protein expression increased, FOXO3a protein relatively decreased the expression of, which in the role of the high dose Chinese traditional medicine group the most obvious (P< 0.05), and the effect is better than the western medicine group, Dingkun Dan group; each drug treatment group differences between no statistical significance (P> 0.05).Each drug treatment group, the expression of HIF-1? protein relative expression amount compared with the model control group were increased, which in the western medicine group, Dingkun Dan group and traditional Chinese medicine high dose group most obvious (P< 0.05), low dose of Chinese medicine group, the difference of no statistical significance (P>0.05); there was no statistically significant difference between the treatment groups.Conclusion:(1) Kidney nourishing kidney and promoting blood circulation traditional Chinese medicine compound kidney pill can effectively shorten the model rats, increase the body weight, ovary wet weight and ovary index, improve serum hormone level, restore the number of follicles and the number of corpus corpus.(2) Induced by Tripterygium Glycosides follicular developmental disorder model rats ovarian tissue existence of transcription factor FOXO3a expression was elevated, HIF-la, IGF-1R expression decreased imbalance; In the model rat of ovarian tissue, Bushen Huoxue compound Newjiaguishen may be through increased IGF-1R level, up regulation of HIF-1? expression and down-regulation of the expression of Pro apoptotic factor FOXO3a, thereby inhibiting excessive follicular atresia and promote follicular growth and development. |
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