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目的观察高糖环境下细胞内硫化氢的生成及硫化氢对高糖诱导的内皮细胞损伤的影响并探讨其机制。方法将人脐静脉内皮细胞分为4组:A组为正常糖浓度组(5.5 mmol/L),B组为高糖组(25 mmol/L),C组为高糖(25 mmol/L)+硫氢化钠(50μmol/L)组,D组为正常糖浓度(5.5 mmol/L)+硫氢化钠(50μmol/L)组,处理48 h后,测定细胞内硫化氢的含量,M TT检测细胞存活率,Western Blot检测细胞凋亡相关蛋白Bax、Bcl-2和Cleaved caspase-3的表达。结果①与A组相比,B组硫化氢生成量明显减少(P<0.05),与B组相比,C组硫化氢生成量明显增多(P<0.05);②与A组相比,B组的细胞存活率明显减少(P<0.05),而C组的细胞存活率较B组显著升高(P<0.05);③与A组相比,B组Bax表达显著升高(P<0.05),Bcl-2的表达显著降低(P<0.05),而Caspase-3活性明显增强(P<0.05);与B组相比,C组Bax水平显著降低(P<0.05),Bcl-2水平显著升高(P<0.05),Caspase-3活性显著下降(P<0.05)。与A组相比,D组各指标表达差异无统计学意义(P>0.05)。结论高糖对内皮细胞硫化氢的生成具有抑制作用;外源性硫化氢可保护高糖诱导的内皮细胞损伤,其机制可能与抑制细胞凋亡有关。
Objective To observe the formation of intracellular hydrogen sulfide under high glucose and the effect of hydrogen sulfide on endothelial cell injury induced by high glucose. Methods Human umbilical vein endothelial cells (HUVECs) were divided into four groups: normal glucose concentration group (5.5 mmol / L), high glucose group (25 mmol / L) + Sodium hydrosulfide (50μmol / L) group, D group was normal glucose concentration (5.5mmol / L) + sodium hydrosulfide (50μmol / L) group. After treated for 48 hours, the content of intracellular hydrogen sulfide was measured. Cell viability was detected by Western Blot. The expressions of Bax, Bcl-2 and Cleaved caspase-3 were detected by Western Blot. Results ① Compared with group A, the formation of hydrogen sulfide in group B was significantly reduced (P <0.05), and the formation of hydrogen sulfide in group C was significantly increased compared with group B (P <0.05); ② Compared with group A, (P <0.05), while the cell viability in group C was significantly higher than that in group B (P <0.05). Compared with group A, the expression of Bax in group B was significantly increased (P <0.05) (P <0.05), while the activity of Caspase-3 was significantly increased (P <0.05). Compared with B group, the Bax level in C group was significantly lower (P <0.05) (P <0.05), while the activity of Caspase-3 decreased significantly (P <0.05). Compared with group A, the expression of each index in group D was not statistically significant (P> 0.05). Conclusion High glucose can inhibit the formation of H2S in endothelial cells. Exogenous hydrogen sulfide can protect endothelial cells induced by high glucose, which may be related to the inhibition of apoptosis.