低分子肝素钙对小鼠胚胎着床的干预作用及机制研究
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(1. 江苏省连云港第一人民医院,江苏连云港 222000; 2. 海南省妇幼保健院生殖医学中心,海口 570206)

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R-33

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Effect and mechanism of low molecular weight heparin calcium on mouse embryo implantation
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(1. The First People’s Hospital of Jiangsu Lianyungang, Lianyungang 222000, China;2. Reproductive Medicine Center, Hainan Maternal and Child Health Care Center, Haikou 570206)

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    摘要:

    目的 探讨低分子肝素钙(Low molecular weight heparin calcium, LMWHC)对小鼠胚胎着床的干预作用及机制?方法 构建小鼠妊娠模型,腹腔注射LMWHC 干预,观察LMWHC 对小鼠胚胎着床的影响?分离培养小鼠滋养层细胞,用不同浓度LMWHC 干预细胞,采用Transwell 小室检测细胞侵袭力,采用ELISA?Western blot和RT-PCR 法检测细胞上清基质金属蛋白酶-2 (matrix metalloproteinases-2, MMP-2) 和金属蛋白酶抑制因子-2(tissue inhibitor of metalloproteinase-2,TIMP-2) 的蛋白和mRNA 水平的表达情况?结果 妊娠小鼠腹腔注射LMWHC 后,妊娠D6 和D9 的小鼠胚胎着床数明显增多( P <0. 05);用1 ×102 IU/ L?1 ×103 IU/ L 和1 ×104 IU/ L 浓度的LMWHC 干预滋养层细胞24 h 后,滋养层细胞侵袭细胞数分别为(84. 83 ± 14. 15)个?(162. 48 ± 25. 49)个?(98. 73 ±12. 77)个,与未用LMWHC 干预细胞的(65. 27 ±13. 92)个比较,差异有显著性( P < 0. 05),滋养层细胞上清液中MMP-2 表达水平较未用LMWHC 干预的细胞均升高( P < 0. 05),滋养层细胞上清液中TIMP-2 表达水平较未用LMWHC 干预的细胞降低( P < 0. 05)?结论 LMWHC 可促进小鼠胚胎着床,其机制可能与LMWHC 通过影响MMP-2 和TIMP-2 表达而增强滋养层细胞侵袭力有关?

    Abstract:

    Objective To explore the effect and mechanism of low molecular weight heparin calcium (LMWHC)on mouse embryo implantation. Methods A mouse pregnancy model was established and LMWC was injected intraperitoneally. The effects of LMWHC on mouse embryo implantation were then observed. Mouse trophoblasts were isolated, cultured, and treated with various concentrations of LMWHC. Transwell chambers were used to examine cell invasiveness. ELISAs, western blotting, and RT-PCR was used to detect the expression levels of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2). Results After intraperitoneal injection of LMWHC into pregnant mice, the numbers of embryo implantations at days 6 and 9 of pregnancy were increased significantly ( P <0. 05).At 24 h after LMWHC injection at 1 × 102 , 1 × 103 , and 1 × 104 IU/ L, invading trophoblast numbers were (84. 83 ±14. 15), (162. 48 ± 25. 49), and (98. 73 ± 12. 77) respectively, significantly different from the cells without LMWHC treatment (65. 27 ± 13. 92) ( P <0. 05). The level of MMP-2 in the culture supernatant of trophoblasts was higher than that in the culture supernatant of cells without LMWHC treatment ( P < 0. 05), while the level of TIMP-2 was lower than that in cells without LMWHC treatment ( P <0. 05). Conclusions LMWHC promotes mouse embryo implantation, and its mechanism may be related to enhancement of the invasiveness of trophoblasts by affecting the expression of MMP-2 and TIMP-2.

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杨文,杨艳,吴晓燕,王芳,孙倩,高原,张欢欢,冯文,马宁.低分子肝素钙对小鼠胚胎着床的干预作用及机制研究[J].中国比较医学杂志,2018,28(12):61~67.

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  • 收稿日期:2018-05-10
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  • 在线发布日期: 2019-01-08
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