【摘要】 目的研究硝酸盐转运蛋白Sialin及编码基因SLC17A5在血管内皮细胞(HUVECs)上的表达和定位及其在血管内皮细胞缺氧复氧损伤后的表达变化。方法应用免疫荧光激光共聚焦成像实验验证Sialin在人脐静脉血管内皮细胞的表达及定位。以SCC-15及人成纤维细胞为参照,通过Western Blot及RT-PCR实验检测Sialin蛋白及编码基因SLC17A5在HUVECs上的表达。HUVECs随机分组,建立不同缺氧损伤时间的血管内皮细胞缺氧复氧损伤模型,通过Real-time PCR实验验证各组SLC17A5基因的表达变化,通过免疫印迹验证各组Sialin蛋白的表达变化。结果 Sialin蛋白的表达位于HUVECs的细胞质与细胞膜,且具有较高的表达水平。血管内皮细胞缺氧复氧损伤后,SLC17A5及Sialin蛋白与对照组相比表达水平均有明显下降(P<0.05),随缺氧时间增加,表达水平继续降低(P<0.05)。结论硝酸盐转运蛋白Sialin在血管内皮细胞的细胞膜及细胞质有明显表达。在血管内皮细胞缺氧复氧损伤中其表达水平与缺氧时间相关。更多还原

【Abstract】 Objective To investigate the expression and location of the nitrate transporter sialin in human vascular endothelial cells with hypoxia/reoxygenation injury(HRI). Methods The expression of SLC17 A5 mRNA and sialin was detected by RT-PCR and Western blot assays. The expression and location of the sialin in HUVEC(human umbilical vein endothelial cells) were observed by immunofluorescence assay with laser scanning confocal microscope. The expression of SLC17 A5 mRNA was examined by Realtime-PCR, and the expression of sialin protein determined by Western blot assay. Results Sialin expressed on cell membrane and in cytoplasm in HUVEC at a relatively high level. The expression of SLC17 A5 and sialin decreased with hypoxia time. Conclusions The nitrate transporter sialin was expressed on cell membrane and in cytoplasm of HUVEC and the expression was related to hypoxia time.更多还原