【摘要】 目的:研究低分子肝素对低氧诱导的早孕细胞滋养细胞凋亡的影响。方法:选取正常早孕(8～10周)绒毛,使用Percoll梯度离心法提取细胞滋养细胞在1%和20%氧分压下分别进行原代培养。两种氧条件下均设对照组与研究组。研究组培养液中加入不同浓度低分子肝素(终浓度为0.1U/ml、1U/ml、10U/ml),未加低分子肝素组作为对照组,将培养孔板分别置入1%和20%氧分压培养箱中培养120h。使用TUNEL法检测细胞凋亡,RT-PCR法检测凋亡基因表达。结果:(1)与20%氧分压条件下对照组相比,在1%氧分压下对照组的细胞滋养细胞的生长明显增加,差异有统计学意义(P<0.05);(2)与对照组比较,1%和20%氧分压下研究组,细胞滋养细胞的生长均明显增加,差异有统计学意义(P<0.05);(3)与20%氧分压条件对照组相比,在1%氧分压下,对照组的细胞滋养细胞的凋亡明显增加,有显著差异(P<0.01);(4)在1%氧分压下,与对照组相比,研究组细胞滋养细胞的凋亡明显减少,差异有统计学意义(P<0.05)。结论:低氧能促进早孕细胞滋养细胞生长,并诱导细胞滋养细胞凋亡;低分子肝素抑制低氧诱导的早孕细胞滋养细胞凋亡并促进其生长。更多还原

【Abstract】 Objective:To investigate the effect of low molecular weight heparin(LMWH) on apoptosis of human primary first trimester cytotrophoblasts induced by hypoxia.Methods:Cytotrophoblast cells were obtained by Percoll gradient centrifugation from early gestational villous tissue(8～10 weeks) and incubated at 1% or 20% oxygen tention respectively.There were control group and study group at two oxygen tention.Different concentrations of LMWH(final concentration of heparin:0.1U/ml,1U/ml,10U/ml) was added to medium of the study group.The control group were added medium without LMWH.The cells were incubated at 1% or 20% oxygen tention for 5 days.Cells number were calculated every 24 hours.TUNEL method was used to determine the growth and apoptosis state of cells.The expressions of Bax and bcl-2 were analyzed by RT-PCR.ANOVA methods and t-test were used for statistic analysis.Results:(1) The number of cytotrophoblasts of the control group increased more obviously at 1% than 20% oxygen tention(P<0.05).(2)LMWH promoted the growth of cytotrophoblasts at 1% and 20% oxygen tension in the study group compared with the control group(P<0.05).(3)Cytotrophoblast apoptosis of the control group increased obviously at 1% compared with 20% oxygen tension(P<0.01).(4)Under the 1% oxygen tention,cytotrophoblast apoptosis reduced obviously at 1% oxygen tension in the study group compared with the control group(P<0.05).Conclusions:Hypoxia can promote the growth of early gestational cytotrophoblasts,but hypoxia can increase the apoptosis of early pregnant cytotrophoblast cells;LMWH can inhibit the apoptosis of cytotrophoblasts induced by hypoxia and promote the growth of early gestational cytotrophoblasts.更多还原