目的观察硫化氢对高糖诱导的血管内皮细胞炎症反应的影响并探讨其机制。方法将人脐静脉内皮细胞(HuVEcs)分为4组:A组为正常糖浓度组(5.5mmol/L),B组为高糖组(16.7mmol/L),C组为高糖(16.7mmol/L)+硫氢化钠(100ixmol/L)组,D组为正常糖浓度(5.5mmol/L)+硫氢化钠(100ixmol/L)组,处理24h后,采用RT—PCR、WesternBlot和电泳迁移率实验(EMSA)检测核因子-KB(NF—KB)的表达和活性;处理48h后,检测环氧合酶-2(COX-2)的表达。结果①与A组相比,B组COX-2蛋白表达量明显增加(P〈0.01),cox-2mRNA表达量明显增加(P〈0.01);与B组相比,c组COX-2蛋白表达量明显降低(P〈0.05),cOx-2mRNA表达量明显降低(P〈0.01)。②与A组相比,B组细胞质内NF—KB蛋白表达量明显降低(JP〈0.05);与B组相比,c组细胞质内NF—KB蛋白表达量明显增加(P〈0.01);与A组相比,B、c、D组NF—KBmRNA表达差异无统计学意义(P〉0.05)。③与A组相比,B组NF—KB活性增加;与B组相比,c组NF—KB活性降低。D组各指标差异无统计学意义(P〉0.05)。结论硫化氢可以抑制高糖诱导的内皮细胞的炎症反应,其机制可能与抑制COX-2的表达有关。
Objective To evaluate the effects of hydrogen sulfide ( H2 S ) on high glucose-induced inflammation of endothelia cells and to explore the possible mechanisms. Methods Human umbilical vein endothelial cells (HUVECs) were randomly divided into 4 groups: group A (5.5 mmol/L glucose), group B ( 16.7 mmol/L glucose), group C ( 16.7 mmol/L glucose + 100 μmol/L NaI-IS) and group D (5.5 mmol/L glucose + 100 p.mol/L NaI-IS). After treating for 24 h, the expression of nuclear factor-KB (NF-KB) was determined by Western blot, mRNA expression was examined by RT-PCR, and the activation was detected by electromobility shift assay (EMSA). After treating for 48 h, protein and mRNA expressions of cyclooxygenase-2 (COX-2) were determined by Western blot and RT-PCR, respectively. Results (1) Compared with that of group A, COX-2 protein expression was distinctly increased( P 〈 0.01 ) and the mRNA level was also increased in group B (P 〈 0.01 ). However, COX-2 protein expression was distinctly decreased( P 〈 0.05 ) as well as mRNA level(P 〈0.01 ) in group C when compared with that of group B. (2) Compared with that of group A, cytoplasmic NF-KB protein expression was obviously decreased in group B ( P 〈 0.05 ), but NariS could stabilize the cytoplasmic NF-KB protein level ( P 〈 0.01 ). There were no significant differences between group A to group D inNF-KB mRNA expression. (3) High glucose increased NF-KB activation in group B. Compared with that of group B, decrease in NF-KB activation was observed in group C. There were no differences between group A and group D in NF- KB and COX-2 expressions and NF-KB activity. Condusion H2S can protect against high glucose-induced inflammation in human umbilical vein endothelial cells, which may be associated with the suppression of COX-2 expression.