环磷酰胺对大鼠卵巢结构与功能的影响及机制  

Effects and mechanism of cyclophosphamide on the ovarian structure and function of rats with premature ovarian failure

作　　者：魏天琴 凌丽[1] 冯秀山[1] 张文倩[1] 熊正爱[1] WEI Tian-qin, LING Li, FENG Xiu-shan, ZHANG Wen-qian, XIONG Zheng-ai(Department of Gynecology and Obstetricsj Second Affiliated Hospitalj Chongcling Medical University, Chongqing 400010, Chin)

机构地区：[1]重庆医科大学附属第二医院妇产科,重庆400010

出　　处：《解放军医学杂志》2018年第3期195-200,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金面上项目(81671415)~~

摘　　要：目的探讨环磷酰胺(CTX)对卵巢结构与功能的影响及其相关机制。方法 50只SD雌鼠随机分为模型组和对照组。模型组大鼠首日以50mg/kg负荷剂量腹腔注射CTX溶液,随后以8mg/(kg·d)维持剂量连续腹腔注射CTX溶液14d建立早发性卵巢功能不全(POI)模型;对照组每日腹腔注射等量生理盐水。观察两组大鼠动情周期、性激素水平及脏器组织形态学改变等指标,判断建模效果;TUNEL法检测卵泡内颗粒细胞的凋亡情况,ELISA检测组织匀浆中相关炎症因子含量,并采用Western blotting检测凋亡相关蛋白的表达水平,初步探讨POI的发生机制。结果与对照组比较,模型组大鼠动情周期紊乱;血清抗苗勒管激素(AMH)、雌激素(E_2)水平明显降低(P<0.01),促卵泡激素(FSH)水平明显升高(P<0.01);各级卵泡数量明显减少(P<0.05);子宫内膜萎缩,子宫壁变薄,腺体稀少。与对照组比较,模型组大鼠卵泡内颗粒细胞凋亡增加,组织匀浆肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)及IL-6含量明显升高(P<0.01),血管内皮生长因子-α(VEGF-α)含量明显降低(P<0.01),凋亡相关蛋白Bax表达水平明显升高(P<0.01),Bcl-2表达水平明显降低(P<0.01)。结论 CTX导致大鼠卵巢结构、生殖及内分泌功能发生明显改变,其机制可能与局部微环境改变及Bax/Bcl-2表达失衡引起颗粒细胞凋亡增加有关。Objective To establish the premature ovarian insufficiency（POI） rat model with cyclophosphamide（CTX）, and investigate the effects and mechanism of CTX on the ovarian structure and function. Methods Fifty female Sprague-Dawley（SD） rats were randomly divided into model group and control group. Rats in model group received intraperitoneal injection of loading dose CTX（50 mg/kg）, and maintenance dose of 8 mg/（kg·d） for 14 days. Rats in the control group were given the same amount of saline during the same period. The modeling effects were judged by observation and comparison of estrous cycle, levels of sex hormone and morphological changes of organs between the two groups; the apoptosis of follicular granulose cell were detected by TUNEL, the contents of inflammatory cytokines in tissue homogenates were detected by ELISA, and the expressions of apoptosis related proteins were measured by Western blotting. Results Compared with control group, the estrous cycle in model group was disordered; the levels of serum anti-mullerian hormone（AMH） and estrogen（E_2） decreased significantly（P0.01） and those of follicle stimulating hormone（FSH） increased markedly（P0.01）; the quantity of follicles decreased obviously（P0.05）; the endometrium atrophied, the uterine wall thinned and the glands decreased in number. Compared with the control group, the apoptosis of follicular granulose cells increased in model group; the contents of TNF-α, IL-1β and IL-6 in the tissue homogenate increased significantly（P0.01）, the contents of vascular endothelial growth factor-α（VEGF-α） decreased markedly（P0.01）; the expression of apoptosis related protein Bax was up-regulated remarkably（P0.01）, while of Bcl-2 was down-regulated obviously（P0.01）. Conclusion CTX may induce the changes of ovary structure and endocrine function, and the mechanism may be related to the local microenvironment changes and the imbalance of Bax/Bcl-2 expression in regulating the apoptosis of granulose cells.

关 键 词：环磷酰胺 早发性卵巢功能不全 动情周期 BAX BCL-2 

分 类 号：R332[医药卫生—人体生理学]