ABSTRACT
Objective To investigate the effect of Jinghou Zengzhi Recipe(JHZZR)on Fas apoptosis pathway in ovarian granulosa cells of rats with controlled ovarian hyperstimulation(COH)and to investigate the possible mechanism of the effect of JHZZR on improving oocyte quality. Methods To establish the model of COH,30 rats were randomly divided into the blank group,the positive group and the TCM group.The expression of Fas,Fasl,Caspase8,Caspase3 in ovarian was detected by qPCR andWestern Blot assay,respectively. Results The expression of Fas,Fasl,Caspase8,Caspase3 was lowered in the blank group and the TCM group. Conclusion JHZZR can inhibit Fas signalling,suggesting that Fas pathway may be one of the mechanisms under-lying that JHZZR improves oocyte quality.
ABSTRACT
Objective To observe the effects of serum containing Jinghou Zengzhi Recipe (JHZZ) on the expression of growth differentiation factor 9 (GDF9) and Bim in cumulus-oocyte complexes (COCs) and cumulus cells (CCs) of controlled ovarian hyperstimulation (COH) rats,and to investigate its curative effect and mechanism.Methods The rat COH ovarian model was prepared for collecting COCs and CCs,which were respectively co-cultured in vitro with the blank serum of female COH SD rat and serum containing JHZZ,and the each group was divided into 3 subgroups:the blank serum group (blank group),serum containing JHZZ group (control group) and serum containing JHZZ plus GDF9 receptor blocker group (experimental group),total 6 subgroups.COCs and CCs were collected after 24 h.The expression levels of GDF9 and Bim mRNA were detected by real-time quantitative PCR.The GDF9 protein expression levels were detected by Western blot.Results (1) The expression level of GDF9 mRNA in control group COCs was obviously higher than that in the control group and experiment group COCs (P＜0.05);the Bim mRNA expression level in COCs of control group and experiment group was significantly lower than that in COCs of blank group (P＜ 0.05);the GDF9 protein expression level in the control group COCs was significantly higher than that in the blank group COCs (P＜0.05),and also higher than that in the experiment group COCs,but the difference was not statistically significant (P＞0.05).(2)The comparison results of GDF9 mRNA expression level in CCs among 3 groups were consistent with those in COCs;the Bim mRNA expression level in CCs of control group and experimental group was significantly lower than that in the blank group (P＜ 0.05);the Bim mRNA expression level in the control group CCs was significantly lower than that in the experimental group CCs (P＜0.05);the GDF9 protein expression level in the control group CCs was significantly higher than that in the blank group and experimental group CCs (P＜0.05).(3)The GDF9 mRNA expression level in the control group COCs was significantly higher than that in CCs (P＜0.05),and the other inter-group comparisons between COSs and CCs had no statistical difference (P＞0.05).Conclusion Serum containing JHZZ can increase the GDF9 expression level in COCs and CCs,maintain the low expression of Bim in COCs and CCs,inhibit the ovarian cells apoptosis and promote the follicular development;COCs is more conducive to the expression of GDF9 mRNA compared with CCs eliminating oocyte.
ABSTRACT
Objective To assess the effects of Jinghou Zengzhi Recipe(JHZZR),a Chinese prescription with the action of tonifying Qi and blood ,on the ovary apoptosis and expression of Bim in ovarian granulosa cells (GCs) of controlled ovarian hyperstimulation(COH) rats , and analyze the possible therapeutic mechanism. Methods A model of COH rats were prepaered and 30 female SD rats were randomly divided into 5 groups , including control group,positive control group,low,medium and high concentration group in six rats in each group. The apoptosis index(AI)in ovarian GCs were detected by TUNEL ,and the expression of Bim by qPCR. Results The AI of ovarian GCs in high and medium concentration group were obviously lower(P 0.05)comparing with control group. The mRNA levels of Bim were lower(P0.05)during the three concentration groups in Bim mRNA. Conclusion JHZZR can inhibit the ovarian GCs apoptosis of COH rats through decreasing the expression of Bim mRNA ,which improve the quality of ovarian follicle.
ABSTRACT
Objective To assess the effects of Jinghou Zengzhi Recipe ,a Chinese prescription with the action of tonifying Qi and blood,on the expressions of Bcl-2,Bax and Caspase3 in ovarian granulosa cells of con-trolled ovarian hyperstimulation(COH)mice. Methods A model of COH mice was prepared and 30 female KM mice were divided into blank group,model group and treatment group. The expressions of Bcl-2,Bax and Cas-pase3 were detected by Real-time Quantitative PCR and Western Blot. Results The mRNA and protein levels of Bax and Caspase3 were lower(P<0.05),while the protein levels of Bcl-2 were significantly higher(P<0.05)in the treatment group than those in the model group. There was no significant differences between the treatment group and blank group in the mRNA and proteins. Conclusion Jinghou Zengzhi Recipe can inhibit the ovarian granulo-sa cellular apoptosis of COH mice through increasing the expression of Bcl-2 protein as well as decreasing the ex-pressions of Bax and Caspase3 mRNA and proteins to nearly the natural level,which improves the quality of ovari-an follicle.