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目的探讨低氧对子痫前期滋养细胞中可溶性血管内皮生长因子受体-1(sFlt-1)和缺氧抑制因子-1α(HIF-1α)的表达及滋养细胞凋亡和侵袭能力的影响。方法取正常妊娠及子痫前期孕妇终止妊娠后的胎盘绒毛组织,分离培养滋养细胞。分为:对照组(正常孕妇):21%O2浓度培养;子痫前期组:分别行21%O2浓度(21%O2组)和1%O2浓度(1%O2组)培养。采用细胞免疫荧光检测滋养细胞密度,实时定量-多聚酶链反应(real tima-PCR)和蛋白印记(Western Blot)法检测滋养细胞中sFlt-1和HIF-1αmRNA及蛋白的表达,TUNNEL法检测滋养细胞调亡,Transwell测定滋养细胞的侵袭能力。结果子痫前期21%O2组、1%O2组与对照组sFlt-1mRNA的表达分别为(2.87±1.94)ng/ml、(3.51±1.25)ng/ml和(1.93±0.77)ng/ml,HIF-1αmRNA的表达分别为(2.79±0.64)ng/ml、(1.77±0.23)ng/ml和(3.63±0.38),三组比较,差异均有统计学意义(P均<0.01)。21%O2组、1%O2组与对照组sFlt-1蛋白分别为(4.99±1.77)μg/ml、(7.02±1.23)μg/ml和(3.83±0.63)μg/ml,HIF-1α蛋白分别为(3.83±0.63)μg/ml、(3.06±0.25)μg/ml和(7.73±1.02)μg/ml,三组比较,差异均有统计学意义(P均<0.01)。21%O2组、1%O2组与对照组72h滋养细胞凋亡分别为(15.81±3.93)个、(20.18±3.47)个和(6.64±1.37)个,侵袭数目分别为(10.05±1.92)个、(18.65±4.17)个和(2.22±0.43)个,三组比较,差异均有统计学意义(P均<0.01)。结论子痫前期和正常妊娠孕妇滋养细胞均对低氧敏感,低氧对滋养细胞sFlt-1和HIF-1α的表达有影响,且低氧条件下促使滋养细胞凋亡及侵袭能力强化。
Objective To investigate the effects of hypoxia on the expression of soluble vascular endothelial growth factor receptor-1 (sFlt-1) and hypoxia-inducible factor-1α (HIF-1α) and the apoptosis and invasion of trophoblast cells in preeclampsia. Methods Placental chorionic villi of pregnant women and pregnant women with preeclampsia after termination of pregnancy were isolated and cultured. Divided into: control group (normal pregnant women): 21% O2 concentration culture; preeclampsia group: 21% O2 concentration (21% O2 group) and 1% O2 concentration (1% O2 group) were cultured. Trophoblast density was detected by immunofluorescence staining. The mRNA and protein expressions of sFlt-1 and HIF-1α in trophoblasts were detected by real-time quantitative polymerase chain reaction (RT-PCR) and Western Blot. TUNNEL assay was used to detect the number of trophoblasts Transplanted, Transwell assay trophoblast invasiveness. Results The expressions of sFlt-1mRNA in 21% O2 group, 1% O2 group and control group were (2.87 ± 1.94) ng / ml, (3.51 ± 1.25) ng / ml and (1.93 ± 0.77) The expression of HIF-1αmRNA was (2.79 ± 0.64) ng / ml, (1.77 ± 0.23) ng / ml and (3.63 ± 0.38) respectively. There were significant differences among the three groups (all P <0.01). The protein levels of sFlt-1 in 21% O2 group, 1% O2 group and control group were (4.99 ± 1.77) μg / ml, (7.02 ± 1.23) μg / ml and (3.83 ± 0.63) μg / ml, (3.06 ± 0.25) μg / ml and (7.73 ± 1.02) μg / ml respectively. There were significant differences among the three groups (all P <0.01). The number of apoptotic cells in 21% O2 group, 1% O2 group and control group at 72 hours were (15.81 ± 3.93), (20.18 ± 3.47) and (6.64 ± 1.37), respectively, and the number of invasiveness was (10.05 ± 1.92) , (18.65 ± 4.17) and (2.22 ± 0.43) respectively. There were significant differences among the three groups (all P <0.01). Conclusion The trophoblastic cells of preeclampsia and normal pregnant women are both sensitive to hypoxia. Hypoxia has effects on the expression of sFlt-1 and HIF-1α in trophoblast cells, and promote the apoptosis and invasion of trophoblast cells under hypoxic conditions.