吕晓丹，李莹，张璐，王学芹，马旭*，夏红飞*

(1.国家卫生计生委科学技术研究所遗传优生中心，北京　100081；2.北京协和医学院研究生院，北京　100730)

化疗药物致卵巢损伤的表型分析及可能机制

吕晓丹1，2，李莹1，张璐1，王学芹1，马旭1*，夏红飞1*

(1.国家卫生计生委科学技术研究所遗传优生中心，北京100081；2.北京协和医学院研究生院，北京100730)

【摘要】目的探索化疗药物对小鼠卵巢的影响及可能的机制。方法将60只ICR小鼠平均分为给药组和对照组。给药组小鼠一次性腹腔注射环磷酰胺120 mg/kg+白消安20 mg/kg，对照组小鼠注射同等体积的DMSO；通过观察小鼠动情周期的变化、性激素水平、卵巢组织形态及卵泡计数分析化疗药物对卵巢的影响；通过检测抗苗勒管激素(AMH)、Ki67表达水平等分析卵巢储备能力和细胞增殖能力的变化。结果给药组雌激素水平(203.00±24.91 pmol/L)比对照组(379.00±36.13 pmol/L)显著降低(P<0.01)；给药组脏器系数(0.0318)比对照组(0.0487)显著下降(P<0.01)；给药组可见卵巢萎缩、各级卵泡数目明显减少、卵泡闭锁增多；给药组颗粒细胞分泌的AMH明显减少、增殖细胞数目显著降低(P<0.01)。结论化疗药物能导致小鼠卵巢损伤，卵巢储备能力和卵巢细胞的增殖能力明显下降。

【关键词】化疗药物；卵巢损伤；表型分析；抗苗勒管激素；增殖

Methods： Sixty ICR mice were equally divided into two groups：the mice were administrated with 120 mg/kg cyclophosphamide and 20 mg/kg busulfan by intraperitoneal injection in CTX+BUS group；and the mice were given with the same quantity of DMSO in the control group. The function of ovary were checked by analyzing estrous cycle，serum gonadal hormone levels，morphology of ovarian tissues，changes of numbers of follicles.Ovarian reserve capacity and cell proliferation were verified by detecting the expression of anti-mullerian hormone and Ki67.

Results： Compared with the control group，the estrogen levels [(203.00±24.91) pmol/L vs. (379.00±36.13) pmol/L] were significantly decreased (P<0.01)；organ coefficient (0.0318 vs. 0.0487) was significantly decreased (P<0.01)；the numbers of follicles reduced and the numbers of atretic follicles increased；AMH levels secreted by granulosa cells significantly reduced and the numbers of proliferating cells were significantly decreased in mice injected with chemotherapy drugs of CTX+BUS group (P<0.01).

Conclusions： Chemotherapy drugs can lead to ovarian damage in mice，and the ovarian reserve capacity and ovarian cell proliferation significantly decreased.

(JReprodMed2016，25(6)：540-544)

化疗药物在抑制肿瘤的同时会产生一定的毒副作用。长期的化疗通常会对女性性腺造成损伤，出现原始卵泡减少等一系列卵巢功能过早衰竭的现象[1-2]。通常卵巢早衰(POF)是指妇女在40岁之前出现闭经、雌激素缺乏(E2<92 pmol/L)、卵泡刺激素(FSH)和黄体生成素(LH)水平升高(FSH>40 U/L，LH>30 U/L)为特征的一种疾病[3-4]。一般人群中发病率为1%～3%，在继发性闭经妇女中占4%～18%，在20岁以前的发病率约为0.01%[5]。目前，如何在化疗的同时保护妇女的卵巢功能、改善其生活质量成为妇产科学及生殖医学研究的重要内容。因此，本实验旨在研究对卵巢毒性较强的化疗药物环磷酰胺(cyclophosphamide，CTX)和白消安(Busulfan，BUS)对小鼠卵巢功能的损伤，为深入探索POF的发病机制和治疗方法提供动物模型和实验依据。