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目的探讨缺氧条件下,Rab11蛋白的表达变化及其在合体滋养细胞胞外体(syncytiotrophoblast exosome,STBEX)过度分泌中的作用。方法将传代培养的Be Wo细胞分为4组(n=24),1Be Wo常氧组:Be Wo细胞在普通培养基、常氧条件下培养;2Be Wo低氧组:Be Wo细胞在普通培养基、低氧条件下培养(92%N2、5%CO2、3%O2);3融合Be Wo常氧组:Be Wo细胞以佛司可林(Forskolin)合体化刺激48 h后,常氧条件下培养;4融合Be Wo低氧组:Be Wo细胞以Forskolin合体化刺激48 h后,低氧条件下培养(92%N2、5%CO2、3%O2)。收集各组细胞上清液并检测STBEX蛋白浓度,采用Western blot检测各组细胞HIF-1α、Rab11蛋白的表达。结果与融合Be Wo常氧组比较,融合Be Wo低氧组上清液中STBEX的蛋白浓度升高,差异具有统计学意义(P<0.05);与Be Wo常氧组比较,Be Wo低氧组上清液中STBEX蛋白浓度升高,差异具有统计学意义(P<0.05);Western blot检测结果:与常氧培养组相比,低氧培养组细胞中HIF-1α与Rab11蛋白表达升高,差异具有统计学意义(P<0.05)。结论缺氧条件下Rab11蛋白可能促进合体滋养细胞胞外体的过度分泌。
Objective To investigate the expression of Rab11 protein and its role in the over-secretion of syncytiotrophoblast exosome (STBEX) under hypoxic conditions. Methods Be Wo cells were subcultured into 4 groups (n = 24) and 1Be Wo normoxia group: Be Wo cells were cultured in normal medium under normoxia; 2Be Wo hypoxia group: Be Wo cells cultured in normal medium (92% N2, 5% CO2, and 3% O2). 3 Beow normoxia group: When Be Wo cells were stimulated by Forskolin for 48 h, 4 fusion Be Wo hypoxia group: Be Wo cells were stimulated with Forskolin for 48 h and cultured under hypoxic conditions (92% N2, 5% CO2, 3% O2). The supernatant of each group was collected and the concentration of STBEX protein was detected. The expression of HIF-1α and Rab11 protein in each group was detected by Western blot. Results Compared with Be Wo normoxia group, the protein concentration of STBEX in supernatants of Be Wo hypoxia group was significantly increased (P <0.05). Compared with Be Wo normoxia group, Be Wo hypoxia Compared with normoxia culture group, the expression of HIF-1α and Rab11 protein in hypoxic culture group was significantly higher than that in normoxia culture group (P <0.05) , The difference was statistically significant (P <0.05). Conclusions Rab11 protein may promote the extracellular secretion of syncytiotrophoblasts in hypoxia.