硫氢化钠对高浓度葡萄糖环境下原代人脐静脉内皮细胞的影响

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5.5mmol/L葡萄糖、25mmol/L葡萄糖、25mmol/L葡萄糖+50μmol/L NaHS及50μmol/L NaHS处理72h，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，25mmol/L葡萄糖组的HUVECs存活率比5.5mmol/L葡萄糖组下降28.37%(P<0.05)，而50μmol/L NaHS和25mmol/L葡萄糖共同处理组的细胞存活率比25mmol/L葡萄糖组上升30.3%(P<0.05)，50μmol/L NaHS组与5.5mmol/L葡萄糖组无明显差异(P>0.05)。与5.5mmol/L葡萄糖组相比，以25mmol/L葡萄糖处理72h后能使CSE蛋白的表达下降(P<0.05)，而25mmol/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25mmol/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。

Effects of sodium hydrosulfide on primary human umbilical vein endothelial cells cultured with high glucose

Objective To observe the effect of sodium hydrosulfide on primary human umbilical vein endothelial cells (HUVECs) cultured with high glucose. Methods The 0.125% trypsin and 0.01% EDTA perfusion method was used to isolate HUVECs. Then, the cells were treated for another 72h with 5.5mmol/L glucose, 25mmol/L glucose, 25mmol/L glucose plus 50μmol/L NaHS or 50μmol/L NaHS. The MTT method was used for cells viability. The protein level of cystathionine gamma-lyase (CSE), a key synthase to produce H2S, was measured by Western blot. Results MTT showed that compared to 5.5mmol/L glucose, cell viability of 25mmol/L glucose group decreased 28.37%(P<0.05). The 25mmol/L glucose plus 50μmol/L NaHS group rose 30.3% (P<0.05) compared to the 25mmol/L glucose group. There was no significant difference between the 50μmol/L NaHS and the 5.5mmol/Lglucose group(P＞0.05). Western blot showed that 25mmol/L glucose could reduce protein expression of CSE(P＜0.05), compared to 5.5mmol/L glucose, and NaHS could ameliorate this phenomenon(P＜0.05). Conclusion High glucose can lower cells viability and decrease expression of CSE in HUVECs and NaHS can ameliorate such effects.