【摘要】 目的 :探讨糖基化终产物 (AGEs)对人脐静脉内皮细胞巨噬细胞炎性蛋白 - 1αmRNA及蛋白表达的影响。方法 :将培养的人脐静脉内皮细胞 (HUVEC)用不同浓度 (10 0mg/L、2 0 0mg/L、4 0 0mg/L)的AGEs孵育 2 4h及同一浓度 (40 0mg/L)AGEs孵育 0h、12h、2 4h及 36h。采用原位杂交方法及Westernblot检测巨噬细胞炎性蛋白 - 1αmRNA及蛋白的表达水平。结果 :BSA组内皮细胞内巨噬细胞炎性蛋白 - 1α呈弱表达 ;10 0mg/L、2 0 0mg/L及 4 0 0mg/LAGEs孵育 2 4h后 ,各组内皮细胞内巨噬细胞炎性蛋白 - 1αmRNA表达的平均积分吸光度值分别为 18 76±3 17、2 6 5 8± 1 6 1及 34 2 3± 2 2 5 (BSA组为 13 83± 1 2 4 ,P <0 0 5 ) ;4 0 0mg/LAGEs孵育 12h、2 4h及 36h后 ,各组内皮细胞内巨噬细胞炎性蛋白 - 1αmRNA表达的平均积分吸光度值分别为 2 2 6 7± 1 4 6、34 2 3± 2 2 5及 4 2 2 8± 3 14 (0h组为 12 5 6± 1 2 4 ,P <0 0 5 )。 10 0mg/L、2 0 0mg/L及 4 0 0mg/LAGEs孵育 2 4h后 ,各组内皮细胞内巨噬细胞炎性蛋白 - 1α蛋白的表达量分别为BSA组的 1 34倍、1 87倍及 2 4 6倍 (P <0 0 5 ) ;4 0 0mg/LAGEs孵育 12h、2 4h及 36h后 ,各组内皮细胞内巨噬细胞炎性蛋白 - 1α蛋白的表达量分别为 0h组的 1 82倍?更多还原

【Abstract】 AIM: To investigate the effects of advanced glycation end products (AGEs) on protein and mRNA expression of macrophage inflammatory protein-1α (MIP-1α) in cultured human umbilical vein endothelial cells(HUVECs). METHODS: HUVECs were cultured with AGEs at different concentrations for 24 h and at a concentration of 400 mg/L for different time.The levels of mRNA and protein expression of MIP-1α in cultured HUVEC were detected by in situ hybridization and Western blot, respectively. RESULTS: In situ hybridization showed that after exposure of HUVECs to AGEs at different concentrations (100 mg/L, 200 mg/L, 400 mg/L) for 24 h, the average integrated optical density values (18.76±3.17, 26.58±1.61, 34.23±2.25) of MIP-1α mRNA expression in HUVECs were higher than that in control group (13.83±1.24, P<0.05). After exposure of HUVECs to AGEs at a concentration of 400 mg/L for 12 h, 24 h and 36 h, the average integrated optical density values of MIP-1α mRNA expression in HUVECs were 22.67±1.46, 34.23±2.25 and 42.28±3.14, higher than that in 0 h group (12.56±1.24, P<0.05). Western Blot showed that exposure of HUVECs to AGEs at different concentrations(100 mg/L, 200 mg/L, 400 mg/L) for 24 h resulted in a 1.34-fold, 1.87-fold and 2.46-fold increase in the expression of MIP-1α protein in HUVECs compared with BSA control group (P<0.05). Meanwhile, exposure of HUVECs to AGEs at a concentration of 400 mg/L for 12 h, 24 h and 36 h resulted in a 1.82-fold, 2.71-fold and 3.34-fold increase in MIP-1α protein expression in HUVECs compared with 0 h group (P<0.05). CONCLUSION: These data suggest that AGEs could induce a high expression of MIP-1α mRNA and protein in cultured HUVECs in a dose-dependent and time-dependent manner.更多还原