Effects Of Baoqing Granule On Sex Hormone Level And Bax,Bcl-2 And VEGF In Rats With Diminished Ovarian Reserve

Objective:To observe the effects of Baoqing Granule on sex hormones for diminished ovarian reserve（DOR）rats and investigate its potential mechanism.Methods:Fifty female SD rats with normal estrus cycle were included in the study.Ten rats were selected as the normal group randomly.The remaining rats were given a single intraperitoneal injection of cyclophosphamide 75mg/kg to induce the DOR model.Thirty-eight rats were divided randomly into the model group,progynova group,Baoqing Granule with high-dose and low-dose group.The rat models in corresponding groups were given saline,progynova（0.21mg/kg）,high-dose（9g/kg）and low-dose（3g/kg）of Baoqing Granule once a day for 3 weeks respectively.The ovarian index was calculated after completed gavaged;Ovarian tissues were taken for morphological observation via HE staining;ELISA was used to determine serum FSH,LH,E₂,AMH and VEGF levels in ovary tissue.The expression of Bcl-2,Bax and VEGF in ovarian tissues was determined by the immunohistochemical method.Results:1.Ovarian wet weight and ovarian index:Compared with the normal group,the ovarian wet weight and ovarian index of the rats in each group were reduced（P<0.05）;compared with the model group,the ovarian wet weight and ovarian index of the rats in each drug intervention group were both increased（P<0.05）;compared with Baoqing Granule high-dose group,the ovarian wet weight and ovarian index in Baoqing Granule low-dose group were reduced,while the ovarian wet weight and ovarian index in progynova group were significantly increased（P<0.05）.2.FSH level:Compared with the normal group,the FSH level of rats in each group receiving CTX were increased,especially model group increased significantly（P<0.05）;compared with the model group,the FSH level decreased in drug intervention groups（P<0.05）;Compared with progynova group,the FSH level in Baoqing low-dose group was still higher（P<0.05）;Compared with Baoqing high-dose group,progynova and Baoqing with low-dose were beneficial to improve FSH level（P>0.05）.3.LH level:Compared with the normal group,the LH level of the Baoqing low-dose group and model group increased significantly（P<0.05）,and the LH level of the Baoqing high-dose group and the progynova group also increased,but there was no statistically difference（P>0.05）;compared with model group,the LH level of each drug intervention group was reduced（P<0.05）;compared with progynova group,the LH level of the Baoqing low-dose group was still higher（P<0.05）;compared with Baoqing high-dose group,Baoqing low-dose group had a poor effect in improving LH level（P<0.05）.4.AMH level:Compared with the normal group,the AMH level of the Baoqing low-dose group and model group decreased significantly（P<0.05）,and the AMH level of Baoqing high-dose and progynova also decreased,but there was no statistical difference（P>0.05）;Compared with model group,the AMH level of each drug intervention groups increased（P<0.05）;compared with progynova group,the AMH level of Baoqing high-dose group increased,and the AMH level of the Baoqing low-dose group decreased,but there was no statistical difference（P>0.05）;compared with Baoqing high-dose group,Baoqing low-dose was not effective in improving AMH level（P<0.05）.5.E₂level:Compared with the normal group,the E₂level of each group was significantly reduced except for the progynova group（P<0.05）;compared with the model group,the E₂level of each drug intervention group were increased（P<0.05）;Compared with progynova group,the E₂level of Baoqing high/low-dose group did not improve significantly（P>0.05）;compared with Baoqing high-dose group,progynova was more effective in improving the E₂level（P<0.05）.6.VEGF level in ovarian tissue:Compared with the normal group,the VEGF level of the model group decreased with statistical significance（P<0.05）;compared with the model group,the VEGF level of each drug intervention group increased（P<0.05）;The remaining groups were compared with each other without significant differences（P>0.05）.7.VEGF protein expression level in ovarian tissue:VEGF protein expression can be seen in the follicles,corpus luteum and granulosa cells of rats in each group.Compared with the normal group,the VEGF protein expression of each group is down-regulated（P<0.05）;Compared with the model group,the expression of VEGF protein in each drug intervention group was up-regulated（P<0.05）;Compared with the progynova group,the expression of VEGF protein in the Baoqing low-dose group was down-regulated（P<0.05）;Compared with Baoqing high-dose group,the VEGF protein expression of Baoqing low-dose group was down-regulated（P<0.05）,and compared with progynova group,there was no significant difference（P>0.05）.8.Bax protein expression in ovarian tissue:Compared with the normal group,the Bax protein expression in the ovarian tissue of each group was up-regulated,and model group was up-regulated significantly（P<0.05）;compared with the model group,Bax protein expressions were all down-regulated in each drug intervention groups,and progynova group was down-regulated significantly（P<0.05）;compared with progynova group,the Bax protein expression in Baoqing high/low-dose groups were both up-regulated（P<0.05）;Baoqing high-dose and low-dose groups were not effective in down-regulating the expression of Bax protein（P<0.05）.9.Bcl-2 protein expression in ovarian tissue:Compared with the normal group,the expression of Bcl-2 protein in the model group and Baoqing low-dose group was down-regulated（P<0.05）;compared with the model group,Bcl-2 protein expression in each drug intervention groups were up-regulated（P<0.05）;compared with progynova group,Bcl-2 protein expression in Baoqing low-dose group was down-regulated（P<0.05）,but there was no significant difference between Baoqing high-dose group（P>0.05）;Compared with Baoqing high-dose group,the Bcl-2 protein expression in Baoqing low-dose group was down-regulated（P<0.05）.Conclusion:CTX had damaged on ovarian reserve,and aggravated follicular apoptosis,increased the expression of Bax protein in ovarian tissue,decreased the expression of VEGF and Bcl-2 protein,increased the levels of serum FSH and LH,and decreased the level of AMH,VEGF and E₂.Baoqing granules can improve the diminished ovarian reserve of CTX model rats.This protective effect may be achieved by reducing the expression of Bax and increasing the expression of VEGF and Bcl-2,thereby reducing serum FSH and LH levels and increasing AMH,E₂and VEGF levels.