化疗药物致卵巢损伤的表型分析及可能机制
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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120mg/kg+白消安20mg/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±24.91pmol/L)比对照组(379.00±36.13pmol/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。
Objective: To investigate the effect and possible mechanism of chemotherapy drugs induced ovarian damage in mice.Methods: Sixty ICR mice were equally divided into two groups:the mice were administrated with 120mg/kg cyclophosphamide and 20mg/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.
Objective: To investigate the effect and possible mechanism of chemotherapy drugs induced ovarian damage in mice.Methods: Sixty ICR mice were equally divided into two groups:the mice were administrated with 120mg/kg cyclophosphamide and 20mg/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.
引文
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[2]刘超英,王哲海,陈黎.肿瘤患者性腺机能的损伤与防治[J].生殖医学杂志,2001,10:242-246.
[3]Goswami D,Conway GS.Premature ovarian failure[J].Hum Reprod Update,2005,11:196-202.
[4]付莉,赵怡璇,李守柔.卵巢早衰实验动物模型的建立[J].生殖医学杂志,2006,15:179-183.
[5]Zargar AH,Salahuddin M,Wani AI,et al.Pregnancy in premature ovarian failure:apossible role of estrogen plus progesterone treatment[J].J Assoc Physicians India,2000,48:213-215.
[6]Wang F,Wang L,Yao X,et al.Human amniotic epithelial cells can differentiate into granulosa cells and restore folliculogenesis in a mouse model of chemotherapy-induced premature ovarian failure[J].Stem Cell Res Ther,2013,4:124.doi:10.1186/scrt335.
[7]Tilly JL.Ovarian follicle counts-not as simple as 1,2,3[J].Reprod Bio Endocrinol,2003,1:1-11.
[8]Visser JA,Durlinger AL,Peters IJ,et al.Increased oocyte degeneration and follicular atresia during the estrous cycle in anti-mullerrian hormone null mice[J].Endorinology,2007,148:2301-2308.
[9]Sowers MR,Eyvazzadeh AD,Mcconnell D,et al.Antimullerian hormone and inhibin B in the definition of ovarianaging and the menopause transition[J].J Clin Endocrinol Metab,2008,93:3478-3483.
[10]Gerdes J,Schwab U,Lemke H,et al.Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation[J].Int J Cancer,1983,31:13-20.
[11]Hemminiki K,Xu G,Angelini S,et al.XPD exon 10and 23polymorphisms and DNA repair in human skin in situ[J].Carcinogenesis,2001,22:1185-1188.
[12]Luisi S,Florio P,Reis FM,et al.Inhibins in female and male reproductive physiology:role in gametogenesis,conception,implantation and early pregnancy[J].Hum Reprod Update,2005,11:123-135.
[13]钱警语,陈秀娟.抗苗勒管激素与女性生育功能关系研究进展[J].生殖与避孕,2013,33:473-478.
[14]王新禹,梁前进.环磷酰胺的毒副作用机制及应对措施[J].药学进展,2006,30:452-456.
[15]Shiraishi A,Sakumi K,Sekiguchi M.Increased susceptibility to chemotherapeutic alkylating agents of mice deficient in DNA repair methyltransferase[J].Carcinogenesis,2000,21:1879-1883.
[16]孙德明,李根平,陈振文,等.实验动物从业人员上岗培训教材[M].北京:中国农业大学出版社,2011:136-141.
[17]俞凌.人脐带间充质干细胞治疗卵巢早衰的初步实验研究[D].北京:解放军总医院军事进修学院,2012:1-87.