Time for preparing a mouse model of autoimmune premature ovarian failure / 中国组织工程研究

ABSTRACT

BACKGROUND: Studies have confirmed that anti-zona zona antibodies can accelerate the destruction and depletion of oocytes, thereby causing premature ovarian failure. OBJECTIVE: To investigate the time of establishing autoimmune premature ovarian failure model in BALB/c mice induced by zona pellucida 3 peptides. METHODS: Thirty healthy female BALB/c mice (7-8 weeks) were randomly divided into immune experimental group (20 mice) and the control group (10 mice). In the experimental group, the mice were given immunization injection starting at 0 week, 0.15 mL of immune reagent injected into the soles at both sides and the lower abdomen. The vaginal exfoliated cell smears were observed every morning to observe the changes in the estrous cycle of the mice. After 2 weeks of injection, 0.15 mL of immune enhancement agent was injected subcutaneously into the same site. On the first day of weeks 4 and 6, the immune agent or immune enhancement agent was injected alternatively. Blood samples were collected before each injection and the serum sex hormone level was measured by enzyme-linked immunosorbent assay. Finally, ovarian tissue and uterus morphology were observed. RESULTS AND CONCLUSION: There was no difference in serum follicle stimulating hormone and estrogen levels between the first immunization injection and 2 weeks after the injection. The serum estrogen level in the experimental group was significantly lower than that in the control group at 4 weeks after injection. The serum estrogen level of the experimental group was significantly lower than that of the control group at 6 weeks after injection (P < 0.05), and meanwhile the serum follicle stimulating hormone level was significantly higher than that of the control group (P < 0.01). The degree of ovarian interstitial fibrosis in the experimental group was more obvious than that in the control group. The ovarian volume decreased and the uterus atrophied in the experimental group. The number of primitive follicles, primary follicles and secondary follicles in mice was significantly lower in the experimental group than the control group, and the number of atresia follicles was significantly higher than the control group (P < 0.05). Therefore, 75 μg of zona pellucida 3 peptides can be used to establish an autoimmune premature ovarian failure disease model in BALB/c mice, and a good modelling effect can be achieved at 6 weeks after immunization.