肿瘤坏死因子α与γ干扰素诱导人绒毛膜滋养层细胞凋亡的研究  

Apoptosis of Cultured Human Trophoblasts Induced by Tumor Necrosis Factor α and Interferon γ

作　　者：邢福祺[1] 孔令红[1] 陈士岭[1] 刘忠[1] 陈东红[1] 

机构地区：[1]第一军医大学附属南方医院妇产科,广州510515

出　　处：《中华妇产科杂志》1999年第8期476-478,I012,共3页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的　探讨肿瘤坏死因子α（ Ｔ Ｎ Ｆα） 与γ干扰素（ Ｉ Ｆ Ｎγ） 诱导妊娠早期人绒毛膜滋养层细胞凋亡的作用及其机理。方法　建立人妊娠早期滋养层细胞体外培养体系； Ｔ Ｎ Ｆα与 Ｉ Ｆ Ｎγ联合作用３６ 小时后，利用台盼蓝染色记数活细胞法检测其细胞毒性。在光镜和透射电镜下观察滋养层细胞凋亡的形态学变化；用琼脂糖凝胶电泳对 Ｄ Ｎ Ａ 断裂片段进行分析；采用流式细胞仪测定滋养层细胞的凋亡指数；粘附式细胞仪检测 Ｔ Ｎ Ｆα作用下滋养层细胞内氧自由基的变化。结果　 Ｔ Ｎ Ｆα在１ ０００ 、３ ０００ 和５ ０００ Ｕ／ｍｌ 浓度时，活细胞数显著减少（ Ｐ＜ ０ ．０５） ；而在与 Ｉ Ｆ Ｎγ共同作用下， Ｔ Ｎ Ｆα在５００ 、１ ０００ 、３ ０００ 和５ ０００ Ｕ／ｍｌ 时，活细胞数均显著减少（ Ｐ＜ ０ ．０５） ；电镜下观察到染色体边集、核固缩、凋亡小体； Ｄ Ｎ Ａ 琼脂糖凝胶电泳有“梯形条带”。细胞凋亡指数具有 Ｔ Ｎ Ｆα浓度的依赖性；在 Ｔ Ｎ Ｆα的作用下，绒毛膜滋养层细胞内氧自由基升高。结论　 Ｔ Ｎ Ｆα与 Ｉ Ｆ Ｎγ能够诱导妊娠早期人绒毛膜滋养层细胞凋亡；氧自由基可能参与了 Ｔ Ｎ Ｆα诱导滋养层细胞凋亡的过程。Objective To investigate the potential role of tumor necrosis factor α (TNF α) and interferon γ (IFN γ) in inducing apoptosis of human trophoblast during early pregnancy and its mechanism. Methods Human trophoblast of early gestation were cultured in vitro. Having cultured with TNF α and IFN γ for 36 hours, their cytotoxic activity was determined by viable cell calculation. Apoptotic morphology was observed under light and electronic microscope. DNA fragment pattern and apoptotic index were analysed by agarose gel electrophoresis and flow cytometer assay (FCM) respectively. Adherent cell analysis and sorting interactive laser cytometer was used to examine the changes of cellular reactive oxygen species (ROS) caused by TNF α. Results TNF α alone significantly decreased the number of viable cells at the concentration of 1 000,3 000 and 5 000 U/ml ( P < 0.05). While combined with IFN γ, TNF α decreased the number of viable cells more markedly. Electronic microscopic examination showed chromatin aggregating beneath the nuclear envelope, nuclear contraction and apoptotic bodies. Agarose gel electophoresis of fragmented DNA showed a ladder like pattern. FCM showed that the apoptotic index of trophoblasts was TNF α concentration dependent. Cellular ROS was increased by TNF α. Conclusion TNF α and IFN γ could induce apoptosis of human trophoblasts during early pregnancy, partly through the mechanism of ROS.

关 键 词：滋养层 绒毛膜 滋养细胞凋亡 Γ干扰素 TNFΑ 

分 类 号：R714.56[医药卫生—妇产科学]