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目的观察硫氢化钠(NaHS,外源性的H2S供体)对高浓度葡萄糖环境下原代人脐静脉内皮细胞(HUVECs)的作用。方法以0.125%胰酶和0.01%EDTA灌注法获得HUVECs,分别用5.5 mmol/L葡萄糖、25 mmol/L葡萄糖、25 mmol/L葡萄糖+50μmol/L NaHS及50μmol/L NaHS处理72 h,采用MTT法检测各组细胞的存活率,Western blot法检测H2S生成酶胱硫醚-γ-裂解酶(cystathionine-γ-Iyase,CSE)的变化。结果 MTT检测表明,25 mmol/L葡萄糖组的HUVECs存活率比5.5 mmol/L葡萄糖组下降28.37%(P<0.05),而50μmol/L NaHS和25 mmol/L葡萄糖共同处理组的细胞存活率比25 mmol/L葡萄糖组上升30.3%(P<0.05),50μmol/L NaHS组与5.5 mmol/L葡萄糖组无明显差异(P>0.05)。与5.5 mmol/L葡萄糖组相比,以25 mmol/L葡萄糖处理72 h后能使CSE蛋白的表达下降(P<0.05),而25 mmol/L葡萄糖+50μmol/L NaHS则能改善这种损害作用(P<0.05),CSE蛋白的表达比25 mmol/L葡萄糖组增强。结论高浓度葡萄糖降低人原代脐静脉内皮细胞的生长和存活率,减少CSE蛋白的表达,而NaHS能够减轻高浓度葡萄糖对CSE的作用。
Objective To observe the effect of sodium hydrosulfide (NaHS) on the primary human umbilical vein endothelial cells (HUVECs) under high glucose concentration. Methods HUVECs were obtained by perfusion with 0.125% trypsin and 0.01% EDTA. The cells were treated with 5.5 mmol / L glucose, 25 mmol / L glucose, 25 mmol / L glucose, 50 μmol / L NaHS and 50 μmol / The survival rate of cells in each group was detected by Western blot. The changes of cystathionine-γ-Iyase (CSE) were detected by Western blot. Results MTT assay showed that the survival rate of HUVECs in 25 mmol / L glucose group decreased by 28.37% (P <0.05) compared with that in 5.5 mmol / L glucose group, while the survival rate of cells treated with 50 μmol / L NaHS and 25 mmol / L glucose group 25 mmol / L glucose group increased by 30.3% (P <0.05). There was no significant difference between 50 mmol / L NaHS group and 5.5 mmol / L glucose group (P> 0.05). Compared with 5.5 mmol / L glucose group, CSE protein expression decreased (P <0.05) at 25 mmol / L glucose for 72 h, while 25 mmol / L glucose + 50 μmol / L NaHS could improve this damage (P <0.05), and the expression of CSE protein was enhanced compared with 25 mmol / L glucose group. Conclusion High concentration glucose can reduce the growth and survival rate of human primary umbilical vein endothelial cells and reduce the expression of CSE protein, while NaHS can reduce the effect of high concentration glucose on CSE.