缺氧再灌注对早孕细胞滋养层细胞生长和凋亡的影响  

Effects of hypoxia/reoxygenation injury on proliferation and apoptosis of human cytotrophoblast cells

作　　者：杨素婷[1] 刘媛[1] 张向丽[1] 李栋[2] 

机构地区：[1]山东大学齐鲁医院妇产科,济南250012 [2]山东大学齐鲁医院低温实验室,济南250012

出　　处：《山东大学学报（医学版）》2011年第12期132-135,138,共5页Journal of Shandong University:Health Sciences

基　　金：山东省博士基金资助项目(2007BS03020)

摘　　要：目的探讨缺氧再灌注对原代培养早孕细胞滋养层细胞生长和凋亡的影响。方法选取正常妊娠早孕(7～9周)绒毛,采用Percoll梯度离心法提取细胞滋养层细胞进行原代培养。将培养细胞随机分为20%氧浓度组(A组)、1%氧浓度组(B组)、缺氧再灌注组(将细胞放入1%氧浓度培养箱2 h后,移入20%氧浓度培养箱培养6 h,C组)。计数细胞数目,采用酶联免疫吸附法(ELISA法)测定肿瘤坏死因子-α(TNF-α)表达,TUNEL法测定细胞凋亡,RT-PCR法检测细胞促凋亡基因Bax、抗凋亡基因Bcl-2及TNF-α基因表达。结果①与A组比较,B组细胞滋养层细胞数目明显增加(P<0.05);与A、B组相比,C组细胞滋养层细胞数目明显减少(P<0.05);②与A组比较,B组细胞培养液中TNF-α浓度明显增加(P<0.05);与B组比较,C组培养液中TNF-α浓度明显增加(P<0.05);③与A组比较,B组凋亡细胞核数目明显增加(P<0.05);与B组相比,C组细胞凋亡明显增加(P<0.05)。结论①低氧促进早孕细胞滋养层细胞生长并诱导其凋亡;②缺氧再灌注抑制早孕细胞滋养层细胞生长并诱导其凋亡。Objective To investigate effects of hypoxia/re-oxygenation（H/R） on proliferation and apoptosis of human primary first trimester cytotrophoblast cells.Methods Cytotrophoblast cells were obtained from early gestational villous tissues by Percoll gradient centrifugation（7-9 weeks）.Control groups A and B were cultured at 1% and 20% O2 for 8 consecutive hours,respectively.The study group C was exposed to H/R（at 1% O2 for 2 hours followed by 20% O2 for 6 hours）.Cells numbers in all groups were calculated after 8 hours.The ELISA method was used to determine tumor necrosis factor-α（TNF-α）.The TUNEL method was used to determine apoptosis of cells.Expressions of Bax,Bcl-2 and TNF-α were analyzed by RT-PCR.Results ① The number of cytotrophoblast cells was obviously increased in group B compared with group A（P0.05）,and cell proliferation in group C was significantly reduced compared with groups A and B（P0.05）.② Concentration of TNF-α in culture medium was significantly increased in group B compared with group A（P0.05）,and it was obviously increased in group C compared with group B（P0.05）.③ Cytotrophoblast cell apoptosis was obviously increased in group B compared with group A（P0.05）,and that in group C was significantly increased when compared with group B（P0.05）.Conclusion Hypoxia can promote growth and increase apoptosis of early pregnant cytotrophoblast cells.H/R can inhibit proliferation and induce apoptosis of early gestational cytotrophoblasts.

关 键 词：缺氧再灌注 细胞滋养层细胞 细胞凋亡 肿瘤坏死因子-Α 

分 类 号：R714.2[医药卫生—妇产科学]