高糖腹膜透析液对HPMECs中GLUT1、SGLT1、TGF-β 1、VEGF-A、CTGF表达的影响研究

Expression of GLUT1,SGLT1,TGFβ-1,VEGF-A,CTGF in human peritoneal microvascular endothelial cells cultured with high glucose peritoneal dialysis fluid

目的探讨高糖腹膜透析液对人腹膜微血管内皮细胞(HPMECs)中GLUT1、SGLT1、TGF-β1、VEGF-A、CTGF表达的影响。方法体外培养HPMECs并分为5组,N组用含葡萄糖的无酚红RPMI-1640培养基培养;P组用2.5%葡萄糖腹膜透析液培养;GN组用含50g/ml葡萄糖转运蛋白1(GLUT1)拮抗剂根皮素的2.5%葡萄糖腹膜透析液培养;SN组用含10g/ml钠-葡萄糖共转运载体1(SGLT1)拮抗剂根皮苷的2.5%葡萄糖腹膜透析液培养;GN+SN组用含50g/ml根皮素和含10g/ml根皮苷的2.5%葡萄糖腹膜透析液培养。分别采用RT-PCR法、Western blot法检测各组HPMECs GLUT1、SGLT1、细胞因子转化生长因子-β1(TGF-β1)、血管内皮生长因子-A(VEGF-A)、结缔组织生长因子(CTGF)的mRNA、蛋白表达水平。结果 5组HPMECs GLUT1、SGLT1、TGF-β1、VEGF-A、CTGF的mRNA、蛋白表达水平比较均有统计学差异(均P<0.05)。与N组比较,P组HPMECs GLUT1、SGLT1、TGF-β1、VEGF-A、CTGF的mRNA、蛋白表达水平均升高(均P<0.05);与P组比较,GN组、SN组、GN+SN组HPMECs GLUT1、SGLT1、TGF-β1、VEGF-A、CTGF的mRNA、蛋白表达水平均降低(均P<0.05)。结论高糖腹膜透析液或通过上调HPMECs中GLUT1、SGLT1的表达,进而上调TGF-β1、VEGF-A、CTGF的表达,从而起促腹膜纤维化作用。

Objective To investigate the effect of high glucose-based peritoneal dialysis fluid（PDF） on the expression of glucose transporters1（GLUT1）,Na＋/glucose cotransporters1（SGLT1）,transforming growth factor-β1（TGF-β1）,vascular endothelial growth factor-A（VEGF-A） and connective tissue growth factor（CTGF） in human peritoneal microvascular endothelial cells（HPMECs）. Methods The cultured HPMECs were divided into 5 groups：normal group（N）,peritoneal dialysis（containing 2.5% glucose） group（P）; GLUT1 antagonists group（GN）,SGLT1 antagonists group（SN）; GLUT1 antagonists＋ SGLT1 antagonists group（GN＋SN）. The mRNA and protein expression of GLUT1,SGLT1,TGF-β1,VEGF-A and CTGF in HPMECs were detected by RT-q PCR and Western blotting,respectively. Results There were significant differences in mRNA and protein expression of GLUT1,SGLT1,TGF-β1,VEGF-A and CTGF among 5 groups（P〈0.05）.Compared with group N,the mRNA and protein expression of GLUT1,SGLT1,TGF-β1,VEGF-A and CTGF in group P was increased significantly（P 〈0.05）. Compared with group P,the mRNA and protein expression of GLUT1,SGLT1,TGF-β1,VEGF-A and CTGF in groups GN,SN and GN＋SN was decreased significantly（P〈0.05）. Conclusion High glucose peritoneal dialysis fluid may up-regulate the expression of GLUT1,SGLT1,which may further induce the up-regulating expression of TGF-β1,VEGF-A and CTGF in human peritoneal microvascular endothelial cells,resulting in the peritoneal fibrosis.