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卵泡刺激素与卵泡刺激素及黄体生成素共同干预对玻璃化冻存小鼠卵巢组织血管内皮生长因子表达的影响
陈杰 常青 杨延周裴秀英 黑常春于佳 孙苗 王燕蓉*
解剖学报 ›› 2015, Vol. 46 ›› Issue (6) : 819-823.
卵泡刺激素与卵泡刺激素及黄体生成素共同干预对玻璃化冻存小鼠卵巢组织血管内皮生长因子表达的影响
Effect of intervention of follicle-stimulating hormone/ follicle-stimulating hormone and luteinizing hormone on the expression of vascular endothelial growth factor in the vitrification mouse ovary
目的 通过在玻璃化冻存全程给予小鼠卵巢组织卵泡刺激素（FSH）及FSH和黄体生成素（LH）共同干预，观察冻融卵巢组织的形态学改变以及两种激素干预对冻存卵巢组织血管内皮生长因子（VEGF）表达的影响，寻找最佳的提高冻融卵巢组织卵泡存活及VEGF表达的激素干预方式。 方法 4周龄C57BL/6J小鼠卵巢组织分为新鲜对照组（CG），玻璃化冻存对照组（VCG），300IU/L FSH全程干预玻璃化冻存组（OG-FSH），以及150IU/L FSH+150IU/L LH全程干预玻璃化冻存组(OG-FSH+LH)，每组30个卵巢样本。通过常规组织学、Western blotting技术，观察并分析各组卵巢组织形态结构改变及VEGF蛋白表达量；通过荧光定量PCR技术(Real-time PCR)检测VEGF mRNA表达情况。 结果 OG-FSH+LH 组正常卵泡百分比最高，且显著高于OG-FSH组（P<0.05）；VEGF蛋白表达量在OG-FSH+LH组显著高于OG-FSH组（P<0.05）；荧光定量PCR检测结果表现为VEGF mRNA表达量在OG-FSH+LH组最高，其次为OG-FSH组，最低是VCG组（P<0.05）。 结论 玻璃化冻存全程添加FSH+LH的干预方式较单独FSH干预具有更高正常卵泡百分比和更佳的VEGF蛋白表达。
Objective To investigate the best way of hormone intervention for protecting the vitrification ovarian tissues via observation of the morphological changes of ovary and the expression of vascular endothelial growth factor (VEGF）after the intervention by follicle-stimulating hormone（FSH）/FSH+ luteinizing hormone (LH）in the whole process of vitrification. Methods Four-week-old C57BL/6J mice were divided into four groups:the fresh control group(CG),the vitrification control group(VCG), the vitrification group intervention by 300 IU/L FSH(OG-FSH) and the vitrification group intervention by 150 IU/L FSH+150 IU/L LH(OG-FSH+LH),30 ovary samples per group. The changes of morphological structure of ovary,the expression of VEGF protein and mRNA in every group were observed and analyzed using routine histology, Western blotting and Real-time PCR. Results The percentage of normal follicles was the highest in OG-FSH+LH group,and significantly higher in OG-FSH+LH group than in OG-FSH group (P<0.05). The expression of VEGF protein was higher in OG-FSH+LH group than OG-FSH group(P<0.05).The order of intensities of expression for VEGF mRNA was OG-FSH+LH group> in OG-FSH group>in VCG group (P<0.05). Conclusion The best intervention way is FSH+LH interpose in the whole process of vitrification, which is in favour of the expression of VEGF.
卵泡刺激素 / 黄体生成素 / 血管内皮生长因子 / 免疫印迹法 / 实时定量聚合酶链反应 / 小鼠
Follicle-stimulating hormone / Luteinizing hormone / Vascular endothelial growth factor / Western blotting / Real-time PCR / Mouse
［1］Meirow D,Ra`anani H,Biderman H.Ovarian tissue cryopreservation and transplantation:a realistic,effective technology for fertility preservation［J］.Human fertility Methods Mol Biol,2014,1154(9):455-743.
［2］Sun FY,Duan XCh,Zhang ZhN, et al.Improved the method of mouce embryos in vitro culture［J］.Acta Anatomica Sinica,2015,46(1):127-132.(in Chinese)
孙芳园,段新崇,张振南,等.小鼠胚胎体外培养方法的改良［J］.解剖学报,2015,46(1):127-132.
［3］De Vos M,Smitz J,Woodruff TK.Fertility preservation in women with cancer［J］. Lancet,2014,384(9950):1302-1310.
［4］Woodruff TK.The emergence of a new interdiscipline: oncofertility［J］.Cancer Treat Res,2007,138(7):3-11.
［5］Treves R,Grynberg M,Parco SI, et al.Female fertility preservation in cancer pationts:an instrumental tool for the envisioning a postdisease life［J］.Future Oncol,2014,10(6):869-874.
［6］Duska LR, Dizon DS. Improving quality of life in female cancer survivors: current status and future question［J］.Future Oncol,2014,10(6):1015-1026.
［7］Aubard Y,Piver P,Cogni Y,et al.Orthotopic and heterotopic autografts of frozen-thawed ovarian cortex in sheep［J］.Hum Reprod,1999,14(8):2149-2154.
［8］Friedman O, Orvieto R, Fisch B, et al. Possible improvements in human ovarian grafting by various host and graft treatments［J］.Hum Reprod,2012,27(2):474-482.
［9］Gamzatova Z,Komlichenko E,Kostareva A, et al. Autotransplantation of cryopreserved ovarian tissue—effective method of fertility preservation in cancer patients［J］.Gynecol Endocrinol, 2014,30(1):43-47.
［10］Shikanov A,Zhang Z,Xu M,et al.Fibrin encapsulation and vascular endothelial growth factor delivery promotes ovarian graft survival in mice［J］.Tissue Eng PartA,2011,17(2324):3095-3104.
［11］Gartrell BA, Tsao CK, Galsky MD. The follicle-stimulating hormone receptor: a novel target in genitourinary malignancies［J］.Urol Oncol,2013,31(8):1403-1407.
［12］Labied S,Delforge Y,Munaut C,et al.Isoform 111 of vascular endothelial growth factor (VEGF111) improves angiogenesis of ovarian tissue xenotransplantation［J］.Transplantation,2013,95(3):426-433.
［13］Wang YR,Chang Q,Sun J,et al. Effects of HMG on revascularization and follicular survival in autotransplant mouse ovarian tissue ［J］.Reprod BioMed Online,2012,24(6):646-653.
［14］Rime H,Nquyen T,Bobe J,et al.Prochloraz-induced oocyte maturation in rainbow trout (Oncorhynchus mykiss).A molecular and functional analysis［J］.Toxicol Sci,2010,118(1):61-70.
［15］Li XY, Kang X, Deng QCh,et al.Combination of a GnRH agonist with an antagonist prevents flare-up effects and protects primordial ovarian follicles in the rat ovary from cisplatin-induced toxicity: a controlled experimental animal study［J］. Reprod Biol Endocrinol,2013,11(16):1-8.
［16］Wang YR,Chang Q, Sun J,et al.Effects of HMG on revascularization and follicular survival in autotransplant mouse ovarian tissue ［J］.Reprod BioMed Online,2012,24(6):646-653.
［17］Li B, Liang HY, Zhou X, et al. Research progress of the expression of gonads FSH and LH receptor［J］. Chnese Journal of Veterinary Science,2009,29(6):816-820. (in Chinese)
李波,梁鸿雁,周虚,等.性腺外FSH受体和LH受体表达研究进展［J］.中国兽医学报,2009,29(6):816-820.
［18］Andersen CY, Ezcurra D.Human steroidogenesis: implications for controlled ovarian stimulation with exogenous gonadotropins［J］. Reprod Biol Endocrinol,2014,115(12):128-129.
［19］Filicori M,Cognigni GE, Samara A, et al.The use of LH activity to drive folliculogenesis: exploring uncharted territories in ovulation induction［J］.Hum Reprod Update,2002,8(6):543-557.
［20］Jeppesen JV, Kristensen SG, Nielsen ME,et al.LH-receptor gene expression in human granulosa and cumulus cells from antral and preovulatory follicles［J］.Clin Endocrinol Metab,2012,97(11):1524-1531.
［21］Jiang YQ, Deng YH.VEGF family and its receptor with retinal neovascularization［J］.International Journal of Ophthalmology,2006,6(5):1113-1116. (in Chinese)
蒋瑶祁,邓耀辉.VEGF家族及其受体与视网膜新生血管形成［J］.国际眼科杂志，2006,6(5):1113-1116.
［22］Shibuya M. Structure and function of VEGF/VEGF-recepter system involve in angiogenesis［J］.Cell Struct Funct，2005,26(1):25-35.
［23］Han H, Silverman F, Santucci T, et al. Vascular endothelial growth factor expression in stage Inon-small cell lung cancer correlates with neoangiongenesis and a poor prognosis［J］.Ann Surg Oncol，2015,8(1):72-79.
［24］Duncan WC, Driesche S, Fraser HM. Inhibition of vascular endothelial growth factor in the primate ovary up-regulates hypoxia-inducible factor-1α in the follicle and corpus luteum［J］. Endocrinology, 2008, 149(7):3313-3320.
促性腺激素干预在卵巢玻璃化冻存中的抗凋亡及促移植后血管重建的作用
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