目的:研究环磷酰胺(CTX)诱导的卵巢早衰(POF)小鼠模型的DNA损伤反应(DDR)。
方法:小鼠注射CTX建立POF模型。尸体,卵巢重量,发情周期,并记录卵巢的病理变化。测定血清中17β-雌二醇(E2)和卵泡刺激素(FSH)水平。Ki67,β-半乳糖苷酶(β-gal)的表达,p21,p53,γH2AX,免疫组织化学检测卵巢组织中的pATM。β-gal的表达,γH2AX,和pATM通过原代培养的颗粒细胞(GC)的免疫荧光染色进行分析。
结果:体重和卵巢重量下降,发情周期不稳定,FSH水平升高,而POF小鼠的E2水平与对照组相比降低。POF的病理结果显示闭锁卵泡增加,黄体,和原始卵泡的数量减少,次要,和三级卵泡。Ki67表达减少,β-gal,p21,p53,γH2AX,pATM表达在POF小鼠卵巢中升高。β-gal的表达,γH2AX,并且pATM在GC中以时间依赖性的方式随浓度增加而增加。
结论:总的来说,CTX诱导小鼠POF,由ATM-P53-P21的DDR通路介导。
OBJECTIVE: To investigate the DNA damage response (DDR) in a cyclophosphamide (CTX)-induced mouse model of premature ovarian failure (POF).
METHODS: The POF model was established by injecting mice with CTX. The body, ovarian weights, the estrus cycle, and pathological changes of the ovaries were recorded. The serum levels of 17 β-estradiol (E2) and follicle-stimulating hormone (FSH) were measured. The expression of Ki67, β-galactosidase (β-gal), p21, p53, γH2AX, and pATM in ovarian tissues was detected by immunohistochemistry. The expression of β-gal, γH2AX, and pATM was analyzed by immunofluorescence staining of primary cultured granulosa cells (GCs).
RESULTS: The body and ovarian weights decreased, the estrus cycles were erratic, and the FSH level increased, whereas the E2 level decreased in POF mice compared to controls. The pathological consequences of POF revealed an increase in atretic follicles, corpus luteum, and primordial follicles and a decrease in the number of primary, secondary, and tertiary follicles. Ki67 expression was reduced, β-gal, p21, p53, γH2AX, and pATM expression were elevated in the ovaries of POF mice. The expression of β-gal, γH2AX, and pATM increased in GCs with the concentration in a time-dependent manner.
CONCLUSIONS: In total, CTX induced POF in mice, which was mediated by the DDR pathway of ATM-P53-P21.