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。
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（P0.05）.（2）LMWH promoted the growth of cytotrophoblasts at 1% and 20% oxygen tension in the study group compared with the control group（P0.05）.（3）Cytotrophoblast apoptosis of the control group increased obviously at 1% compared with 20% oxygen tension（P0.01）.（4）Under the 1% oxygen tention,cytotrophoblast apoptosis reduced obviously at 1% oxygen tension in the study group compared with the control group（P0.05）.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.
出处
《现代妇产科进展》
CSCD
北大核心
2010年第5期350-354,共5页
Progress in Obstetrics and Gynecology
基金
山东省博士基金资助项目(No:2007BS03020
2007BS03052)
