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  

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

作　　者：刘文[1] 高聆[2] 于春晓[1] 王佳[3] 范右飞[1] 张媛[1] 管庆波[1] 赵家军[1] 

机构地区：[1]山东大学附属省立医院内分泌科,山东省临床医学研究院内分泌代谢研究所,山东省内分泌代谢病临床医学中心 [2]山东大学附属省立医院中心实验室 [3]山东大学附属省立医院妇产科,济南250021

出　　处：《山东大学学报（医学版）》2011年第7期15-18,23,共5页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助项目(30971408);山东省自然科学基金资助项目(Y2008C113)

摘　　要：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 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 72 h with 5.5 mmol/L glucose,25 mmol/L glucose,25 mmol/L glucose plus 50 μmol/L NaHS or 50 μmol/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 5.5 mmol/L glucose,cell viability of 25 mmol/L glucose group decreased 28.37%（P〈0.05）.The 25 mmol/L glucose plus 50 μmol/L NaHS group rose 30.3%（P〈0.05） compared to the 25 mmol/L glucose group.There was no significant difference between the 50 μmol/L NaHS and the 5.5 mmol/L glucose group（P〈0.05）.Western blot showed that 25 mmol/L glucose could reduce protein expression of CSE（P〈0.05）,compared to 5.5 mmol/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.

关 键 词：硫氢化钠 高浓度葡萄糖 胱硫醚-Γ-裂解酶 原代人脐静脉内皮细胞 

分 类 号：R587.1[医药卫生—内分泌]