摘要
目的观察低分子肝素及肝素结合型表皮生长因子（HB-EGF）对孕早期滋养细胞功能的影响。方法2011年2月至11月在中山大学附属第二医院进行体外培养孕早期绒毛组织中的滋养细胞。按照不同浓度的低分子肝素分为0．025U／ml组、0．25U／ml组、2．5U／ml组、25U／ml组、250U／ml组；按照低分子肝素、HB．EGF单独及联合用药，分为低分子肝素组（浓度0．25U／m1），HB-EGF组（浓度10μg／L），联合用药组（浓度0．25U／ml的低分子肝素＋10μg/L的HB-EGF）；将加入DMEM设为对照组。采用四甲基偶氮唑蓝比色法检测孕早期滋养细胞增殖能力，以平均吸光度（A）值表示；采用穿膜小室实验检测孕早期滋养细胞的侵袭能力，以穿透至膜下细胞数表示；并检测培养上清液中孕早期滋养细胞分泌的hCG水平以评价滋养细胞的分化能力。结果与对照组比较，低分子肝素浓度为0．025U／ml对孕早期滋养细胞增殖、侵袭能力影响不明显（P〉0．05）；低分子肝素浓度为0．25U／ml、2．5U／ml时，低分子肝素增加孕早期滋养细胞增殖、侵袭能力（P〈0．05）；低分子肝素浓度为25U／ml、250U／ml时，则低分子肝素显著抑制孕早期滋养细胞增殖、侵袭能力（P〈0．05）。与对照组A值（0．44±0．04）比较，低分子肝素组（A值为0．51±0．05）、HB-EGF组（A值为0．56±0．04）、联合用药组（A值为0．69±0．06）均显著增高（P〈0．05）。在侵袭试验中，低分子肝素组侵袭细胞数为（511±78）个，HB-EGF组为（669±67）个，联合用药组为（872±64）个，分别与对照组（405±67）个比较，差异均有统计学意义（P〈0．05）。低分子肝素组上清液中hCG水平为（7143±649）U／L，HB-EGF组为（11762±1059）U／L，联合用药组为（11015±1084）U／L，分别与对照组（8182±666）U／L比较，差异也均有统计学意义（P〈0．05）。结论低
Objective To evaluate the effects of low molecular weight heparin （LMWH） and heparin-binding epidermal growth factor （HB-EGF） on the biological function of human trophoblast in first trimester. Methods From Feb. 2011 to Nov. 2011, the trophoblast isolated from human first trimester chorionic villi was cultured in vitro. Based on variation of LMWH concentration, the trophoblast was classified into 0. 025 U/ml group, 0. 25 U/ml group, 2. 5 U/ml group, 25 U/ml group and 250 U/m1 group. In the mean time, based on treatment of heparin, the trophoblast was classified into LMWH group （0. 25 U/ml）, HB-EGF group （10 μg/L）, combination group （LMWH at 0. 25 U/ml ± HB-EGF at 10 μg/L） and add with DMEM as control group. Cell proliferation was assessed by the methyl thiazolyl tetrazolium （MTT） test, which was showed with the mean absorbance as A value. Cell invasion was measuredby transwell, which counted the number of cells migrated to the superficies inferia of filter membrane. Cell differentiation was assessed by the concentration of hCG secretion. Results Compared with control group, the trophoblast proliferation and invasion treated by LMWH at 0. 025 U/ml did not show significant difference （P 〉 0. 05）. When treated by LWMH at 0. 25 U/ml and 2.5 U/ml, trophoblast proliferation and invasion was increased significantly （P 〈 0.05 ）. When LMWH at 25 U/ml and 250 U/ml, it could inhibit trophoblast proliferation and invasion （ P 〈 0. 05 ）. When compared with A value of 0. 44 ± 0. 04 in control group, the increased A value were 0. 51 ±0. 05 in LMWH group, 0. 56 ± 0. 04 in HB-EGF group and 0. 69 ± 0. 06 in combination group （P 〈 0. 05 ）. In the transwell test, the cell number were 511 ± 78 in LMWH group, 669 ± 67 in HB-EGF group and 872 ± 64 in combination group, which were significantly higher than 405 ± 67 in control group （ P 〈 0.05 ）, respectively. And the hCG concentration were （ 7143 ± 649 ） U/L in LMWH group, （11 762 ± 1059） U/L in HB-EGF group and
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2013年第2期107-112,共6页
Chinese Journal of Obstetrics and Gynecology
基金
国家自然科学基金（81170625）,教育部高等学校博士学科点专项科研基金（20090171120075）
关键词
肝素
低分子量
胞间信号肽类和蛋白质类
滋养层
妊娠初期
细胞增殖
细胞分化
Heparin, low-molecular-weight
Intercellular signaling peptides and proteins
Trophoblasts
Pregnancy trimester, first
Cell proliferation
Cell differentiation
