低氧预处理人脐带间充质干细胞治疗早发性卵巢功能不全小鼠的效果研究
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摘要
目的探索人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUCMSCs)及其经低氧预处理后对早发性卵巢功能不全(premature ovarian insufficiency,POI)小鼠的治疗作用。方法从脐带中分离、培养出hUCMSCs,利用白消安和环磷酰胺诱导形成小鼠POI模型,分别使用正常hUCMSCs和经低氧预处理的hUCMSCs对POI小鼠进行治疗。实验结束时,测量各组(n=15)小鼠血浆中雌二醇(estradiol,E2)、卵泡刺激素(follicle stimulating hormone,FSH)及抗苗勒管激素(anti Mullerian hormone,AMH)浓度,对卵巢进行组织病理学检查,计数卵泡数量。结果经过hUCMSCs和低氧预处理hUCMSCs治疗的小鼠血浆E2及AMH浓度,与模型对照组比较,呈升高趋势;且AMH浓度差异有统计学意义(P <0. 05);与模型对照组比较,hUCMSCs和低氧预处理hUCMSCs治疗组小鼠卵巢组织中生长卵泡明显增多、闭锁卵泡明显减少(P <0. 05);卵巢颜色红润,尺寸增大,几乎没有坏死点,卵泡大、透明度好,整体来看与正常对照组卵巢状态相似;且经低氧预处理hUCMSCs治疗的小鼠,其生长卵泡和闭锁卵泡数量更趋于正常。结论 hUCMSCs移植及hUCMSCs低氧预处理后移植治疗均能够有效恢复POI小鼠的激素浓度,促进卵巢功能的恢复。
Objective To investigate the therapeutic effect of hypoxia-preconditioned human umbilical cord mesenchymal stem cells(hUCMSCs) on premature ovarian insufficiency(POI) mice model. Methods hUCMSCs were isolated and cultured from umbilical cord,and POI mice model were induced by busulfan and cyclophosphamide. POI mice were treated with normal hUCMSCs and hypoxic pretreated hUCMSCs. At the end of the experiment,the levels of estradiol(E2),follicle stimulating hormone(FSH) and anti Mullerian hormone(AMH) in the plasma were measured,histopathological examination was performed on ovaries,and the number of follicles was counted. Results Plasma E2 and AMH levels in mice treated with hUCMSCs and hypoxic preconditioned hUCMSCs showed an increasing trend,with a significantly different in AMH levels(P < 0. 05) compared with the control group. The growth of follicles in the ovarian tissue of the hUCMSCs and hypoxic preconditioned hUCMSCs groups was significantly increased,and the atresia follicles were significantly decreased(P < 0. 05). The color of the ovary was ruddy,and the size was increased,with almost no necrosis. The follicles were large,with good transparency. The ovarian status of the normal control group was similar,and the number of growing follicles and atresia follicles in the treated groups became more normal. Conclusion hUCMSCs transplantation significantly improved the serum levels of high gonadotropin and low estrogen of POI mice,and promote follicular development.
Objective To investigate the therapeutic effect of hypoxia-preconditioned human umbilical cord mesenchymal stem cells(hUCMSCs) on premature ovarian insufficiency(POI) mice model. Methods hUCMSCs were isolated and cultured from umbilical cord,and POI mice model were induced by busulfan and cyclophosphamide. POI mice were treated with normal hUCMSCs and hypoxic pretreated hUCMSCs. At the end of the experiment,the levels of estradiol(E2),follicle stimulating hormone(FSH) and anti Mullerian hormone(AMH) in the plasma were measured,histopathological examination was performed on ovaries,and the number of follicles was counted. Results Plasma E2 and AMH levels in mice treated with hUCMSCs and hypoxic preconditioned hUCMSCs showed an increasing trend,with a significantly different in AMH levels(P < 0. 05) compared with the control group. The growth of follicles in the ovarian tissue of the hUCMSCs and hypoxic preconditioned hUCMSCs groups was significantly increased,and the atresia follicles were significantly decreased(P < 0. 05). The color of the ovary was ruddy,and the size was increased,with almost no necrosis. The follicles were large,with good transparency. The ovarian status of the normal control group was similar,and the number of growing follicles and atresia follicles in the treated groups became more normal. Conclusion hUCMSCs transplantation significantly improved the serum levels of high gonadotropin and low estrogen of POI mice,and promote follicular development.
引文
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[13]Lai D,Wang F,Yao X,et al.Human endometrial mesenchymal stem cells restore ovarian function through improving the renew al of germline stem cells in a mouse model of premature ovarian failure[J].J Transl M ed,2015,13:155.
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[16]Sun L,Li D,Song K,et al.Exosomes derived from human umbilical cord mesenchymalstem cells protect against cisplatin-induced ovarian granulosa cell stress and apoptosis in vitro[J].Sci Rep,2017,7(1):2552.
[17]Han K H,Kim A K,Kim M H,et al.Enhancement of angiogenic effects by hypoxia-preconditioned human umbilical cord-derived mesenchymal stem cells in a mouse model of hindlimb ischemia[J].Cell Biol Int,2016,40(1):27-35.
[18]Choi J R,Yong K W,Wan Safwani W K Z.Effect of hypoxia on human adipose-derived mesenchymal stem cells and its potential clinical applications[J].Cell M ol Life Sci,2017,74(14):2587-2600.
[2]Shelling A N.Premature ovarianfalure[J].Reproduction,2010,140(5):633-641.
[3]Thakur M,Feldman G,Puscheck E E.Primary ovarian insufficiency in classic galactosemia:current understanding and future research opportunities[J].J Assist Reprod Genet,2018,35(1):3-16.
[4]Jankowska K.Premature ovarian failure[J].Prz Menopauzalny,2017,16(2):51-56.
[5]Fazeli Z,Abedindo A,Omrani M D,et al.Mesenchymal stem cells(M SCs)therapy for recovery of fertility:a systematic review[J].Stem Cell Rev,2018,14(1):1-12.
[6]Paladino F V,Peixoto-Cruz J S,Santacruz-Perez C,et al.Comparison betw een isolation protocols highlights intrinsic variability of human umbilical cord mesenchymal cells[J].Cell Tissue Bank,2016,17(1):123-136.
[7]Buravkova L B,Andreeva E R,Gogvadze V,et al.Mesenchymal stem cells and hypoxia:w here are w e[J].M itochondrion,2014,19 Pt A:105-112.
[8]Li J,Mao Q,He J,et al.Human umbilical cord mesenchymal stem cells improve the reserve function of perimenopausal ovary via a paracrine mechanism[J].Stem Cell Res Ther,2017,8(1):55.
[9]Kovanci E,Schutt A K.Premature ovarian failure:clinical presentation and treatment[J].Obstet Gynecol Clin North Am,2015,42(1):153-161.
[10]Xiang J,Jiang T,Zhang W,et al.Human umbilical cordderived mesenchymal stem cells enhanced HK-2 cell autophagy through M icroRNA-145 by inhibiting the PI3K/AKT/mTOR signaling pathw ay[J].Exp Cell Res,2019,378(2):198-205.
[11]Song D,Zhong Y,Qian C,et al.Human umbilical cord mesenchymal stem cells therapy in cyclophosphamide-induced premature ovarian failure rat model[J].Biomed Res Int,2016,2016∶2517514.
[12]Liu S,Wang J,Han R.Therapeutic effect of transplanted umbilical cord mesenchymal stem cells in a cynomolgus monkey model of multiple sclerosis[J].Am J Transl Res,2019,11(4):2516-2531.
[13]Lai D,Wang F,Yao X,et al.Human endometrial mesenchymal stem cells restore ovarian function through improving the renew al of germline stem cells in a mouse model of premature ovarian failure[J].J Transl M ed,2015,13:155.
[14]Deryabin P,Griukova A,Shatrova A,et al.Optimization of lentiviral transduction parameters and its application for CRISPR-based secretome modification of human endometrial mesenchymal stem cells[J].Cell Cycle,2019,18(6-7):742-758.
[15]Shi Q,Gao J,Jiang Y,et al.Differentiation of human umbilical cord Wharton's jelly-derived mesenchymal stem cells into endometrial cells[J].Stem Cell Res Ther,2017,8(1):246.
[16]Sun L,Li D,Song K,et al.Exosomes derived from human umbilical cord mesenchymalstem cells protect against cisplatin-induced ovarian granulosa cell stress and apoptosis in vitro[J].Sci Rep,2017,7(1):2552.
[17]Han K H,Kim A K,Kim M H,et al.Enhancement of angiogenic effects by hypoxia-preconditioned human umbilical cord-derived mesenchymal stem cells in a mouse model of hindlimb ischemia[J].Cell Biol Int,2016,40(1):27-35.
[18]Choi J R,Yong K W,Wan Safwani W K Z.Effect of hypoxia on human adipose-derived mesenchymal stem cells and its potential clinical applications[J].Cell M ol Life Sci,2017,74(14):2587-2600.