Effects Of Xinjiaguishen Pill On P-Akt, XIAP And Apaf-1 In Rats With Follicular Developmental Disorders Induced By Tripterygium Glycosides

OBJECTIVE Based on the rat model of follicular maldevelopment induced by GTW,The experiment is to study the expression of p-Akt.XlAP.Apaf-land PCNA.as well as the relationship between XIAP.Apaf-1 and the P13K/Akt signaling pathway on these animal models treated by Compound Herbal of Xinjiaguishenwan.and also to explore the molecular mechanism of Compound Herbal of Xinjiaguishenwan with the function of Tonifying Kidney and Activating Blood on promoting the development follicle from two aspects of cell proliferation and apoptosis.METHODS Reference to the previous experimental study.we builded rat models of follicular maldevelopment required in the experiment by given intra-gastric administration with GTW 75mg/(kg-d) for 30 days.Then these rats were randomly divided into 5 groups.namely:the model control group.two different Chinese herb dosages(high and low)groups treated by herbal compound of Xinjiaguishenwan.the western medicine control group and the Dingkundan group, together with the normal control group which randomly selected before.there were 6 groups in total and each group consisting of 10 rats. The two Chinese herb groups were treated with two different dosages of herbal compound of Xinjiaduishenwan through intra-gastric administration;The western medicine group were treated with estrogen and progestin sequentially;The Dingkundan group were given Dingkundan water pill solution;The two control groups were both daily given distilled water by gastric perfusion.All groups were continuously treated for 2 weeks.During the treatment,Vaginal exfoliated cells smears was taken.Femoral artery blood was collected to detect the concentration of serum "E2""FSH"with RIA,ovaries of each rat was taken out from the abdominal cavity to scale the weight of the ovaries and calculate the ovarian index.Count the primordial, atscondary, antral, resia follicles and corpus luteum by optical microscope on the stained paraffin sections of ovarian with HE.Observing the gene and protein expression of "p-Akt""XIAP""Apaf-r'and "PCNA" in ovaries of the rats with Western blotting and RT-PCR respectively.Analysing the correlation between "p-Akt”and "XIAP"" Apaf-1'RESULTS GTW 75mg/(kgd) gavaged for 30 days can result in follicular maldevelopment on the child-bearing period femal SD rats model. ? Compared with the normal group, the oestrous cycle of model rats were prolonged, disordered or even disappeared.the weight.ovarian index and serum E2 level were declined.serum FSH were increased.The percentage of growing follicles and the number of corpus luteum decreased, the percentage of atresia follicles increased. Ovarian PCNA, p-Akt. XIAP levels were decreased, Apaf-1 expression increased, the differences were statistically significant(P<0.01 or P<0.05).?Compared with the model control group, the oestrous cycle of the Chinese medicine groups and the western medicine group were shortened.the weight.ovarian index and serum E2 level were increased,serum FSH were decreased.The percentage of growing follicles and the number of corpus luteum were increased, the percentage of atresia follicles were declined. Ovarian PCNA. p-Akt. XIAP levels were increased. Apaf-1 expression decreased.the differences were statistically significant of all above indictors(P<0.01or P<0.05).The results of high dosage of Herbal compound group and the western medicine group were more obvious than low dosage.?p-Akt showed a significant positive correlation on PCNA(i=0.763,P=0.00);XIAP was positively correlated with p-Akt(r=0.664,P=0.00);Apaf-1 was negatively correlated with p-Akt(r= 0.663.P=0.00).CONCLUSION GTW probably inhibit the activity of PI3K/Akt signaling pathway through down-regulating the expression of p-Akt which is a key molecule of that signaling pathway to influnce the expression of XIAP and Apaf-1 in every rat ovary.thus block the normal growth, development and maturity of follicles. However,the Herbal of Xinjiaguishenwan with the functions Tonifying Kidney and Circulating Blood may repair the activity of PI3K/Akt signaling pathway via up-regulating the expression of p-Akt,besides Up-regulating the anti-apoptotic factor XIAP, down-regulating Pro-apoptotic factor Apaf-1 to relieve the inhibitory effect of GTW.therefore promoting the normal development of follicles, restoring the disordered function of the ovaries.