| Objective: To establish a rat model of premature ovarian failure combined with chronic psychological stress by intragastric administration of Tripterygium wilfordii glycosides and chronic unpredictable mild stimulation in rats,and to explore the effect of Guiluo’s Anzang Decoction on ovarian reserve function in rats with premature ovarian failure combined with chronic psychological stress and its mechanism.Methods: Through open field experiment and vaginal exfoliative cell smear observation,rats with similar behavioral scores and normal estrous cycle were selected from the purchased female SD rats to enter the experiment.Six of them were randomly selected as the blank group and fed freely.A daily dose of 10 ml · kg-1 of physiological saline was administered by gavage for 35 days.The remaining rats were included in the model preparation group and administered a daily dose of 75 mg · kg-1 of Tripterygium wilfordii polyglycoside tablet suspension for14 consecutive days to establish a rat model of premature ovarian failure.After the completion of modeling,30 rats in the model preparation group were selected and randomly divided into model group,positive group(estradiol valerate tablets),high,medium,and low dose groups of Guiluoshi Anzang Tang,a total of 5 groups,with 6 rats in each group.The rats in each group were given corresponding therapeutic drugs and simultaneously subjected to chronic unpredictable mild stimulation.The intervention time was 21 days.After the intervention,the behavioral scores of rats were recorded through open field test(OFT),and the pathological morphology of ovarian tissue was observed by HE staining;Serum levels of follicle stimulating hormone(FSH),estradiol(E2),and anti muscleus detonator hormone(AMH)were detected by enzyme linked immunosorbent assay(ELISA);Real-time PCR was used to detect the expression levels of PI3 K,AKT,FOXO3 a,and Caspase3 m RNA in ovarian tissue;Western blot was used to detect the expression levels of PI3 K,AKT,FOXO3 a,and Caspase3 proteins in ovarian tissue.Result:1.Ovarian histomorphology observation: Compared with the blank group,the number of follicles and corpus luteum at all levels in the model group and the treatment group decreased,while the number of atresia follicles increased.Compared with the model group,the ovarian tissue morphology in the treatment group was improved to varying degrees.2.Serum FSH,E2,and AMH levels: Compared with the blank group,FSH levels in the model group increased(P<0.05),while E2,and AMH levels decreased(all P<0.05).Compared with the model group,the FSH level in each treatment group decreased to varying degrees,with the most significant statistical difference between the positive group and the high-dose group of traditional Chinese medicine(all P<0.05);E2 and AMH levels increased to varying degrees,with the most significant difference between the positive group and the high-dose group of traditional Chinese medicine(both P<0.05).3.Expression of PI3 K,AKT,FOXO3 a,Caspase3 m RNA and protein in ovarian tissue: In this experiment,there was no significant difference in the expression of PI3 K,AKT m RNA and protein in ovarian tissue of rats in each group(all P>0.05).Compared with the blank group,the expression of FOXO3 am m RNA and protein in the model group decreased(P<0.05),while the expression of Caspase3 m RNA and protein increased(P<0.05).Compared with the model group,the expression of FOXO3 am m RNA and protein in each treatment group increased to varying degrees,with the most significant difference between the positive group and the high-dose group of traditional Chinese medicine(both P<0.05).Compared with the model group,the expression of caspase3 m RNA and protein in each treatment group decreased to varying degrees,with the most significant differences in the positive group,the high-dose group of traditional Chinese medicine(all P<0.05).4.Open field experiment results: In this experiment,there was no significant difference in the total horizontal movement distance of rats in each group(all P>0.05).Compared with the blank group,the number of times the model group crossed the central grid decreased(P<0.05).Compared with the model group,the number of times each treatment group crossed the central grid increased to varying degrees,with the most significant difference in the high-dose group of traditional Chinese medicine(P<0.05).Conclusion:1.A rat model of premature ovarian failure combined with chronic psychological stress can be established by intragastric administration of Tripterygium wilfordii polyglycoside tablets combined with chronic unpredictable mild stimulation.2.Guiluoshi Anzang Decoction can improve the ovarian reserve function and anxiety like behavior in chronic psychological stress model rats with premature ovarian failure.3.Guiluoshi Anzang Decoction can increase the expression level of FOXO3 am m RNA and protein in ovarian tissue of rats with premature ovarian failure and chronic psychological stress;Reduce the expression level of Caspase3 m RNA and protein in ovarian tissue,and inhibit apoptosis in ovarian tissue. |
PDF Full Text Request