Objective: To compare the methods of establishing animal models of premature ovarian failure (POF) induced by cyclophosphamide (CPF) in rabbits under different injection methods, and to find the best way of modeling, so as to provide more suitable experimental models for further research on POF?
Methods: Twenty one rabbits were randomly divided into four groups. Three rabbits were in the normal control group without any treatment (group A); Six rats in model group I were injected with 50 mg/kg cyclophosphamide solution for 1 day (group B); 6 rats in model group II were injected with 50 mg/kg for 2 days (group C); Six rats in three groups of models were injected with 50 mg/kg on the first day, and then injected with 8 mg/kg continuously for 14 days (Group D)? Weigh the weight and ovary weight of rabbits in each group, describe the weight change within 14 days after injection, and analyze the ovary index; The morphology of follicles was observed by HE staining, and the number of follicles at all levels was calculated under light microscope; TUNEL method was used to observe and analyze the apoptosis; Analysis of serum estradiol (E2) changes by ELISA?
Result: Body weight: B? There was no significant change in body weight in group C (P>0.05); The growth of rabbits in group D was restricted (P<0.05), and the mortality was high? Ovarian index: Ovarian index in group C was significantly lower than that in group A (P<0.05)? Follicles at all levels: B? The abnormal rate of primordial follicles and primary follicles in group C increased (P<0.017), and the abnormal rate of primordial follicles in group C increased more significantly (P<0.017), but="" there="" was="" no="" significant="" difference="" between="" the="" two="" groups="" in="" abnormal="" rate="" of="" primary="" follicles="" p="">0.017)? A? B? C Three groups of secondary? There was no significant difference in the abnormal rate of sinus follicles (P>0.05)? Apoptosis rate: Apoptosis mainly occurs on granulosa cells in follicles? The apoptosis rate of group C was significantly higher than that of group A (P<0.05), and the apoptosis rate of group C was higher than that of group B (P<0.05)? Serum E2 value: In group B, the E2 secretion reached the peak on the 7th day and gradually decreased after the 7th day, and the E2 secretion was significantly higher on the 7th day than on the 35th day (P<0.05); The E2 value of group C continued to decline, and was significantly higher on day 0 than on day 35 (P<0.05)?
Conclusion: 50 mg/kg continuous injection for 2 days is the best way to establish a rabbit model of premature ovarian failure induced by cyclophosphamide?