CSEsiRNA对EAhy926细胞的CSE及H_2S的表达影响  

The effects of CSEsiRNA on the expression of CSE and H_2S in EAhy926 cells

作　　者：孙璇君[1] 胡友洋[1] 陈志武[1] 

机构地区：[1]安徽医科大学药理学教研室,合肥230032

出　　处：《安徽医科大学学报》2012年第8期904-907,共4页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金项目(编号:81173596);安徽省自然科学基金项目(编号:11040606M196)

摘　　要：目的观察胱硫醚γ裂解酶(CSE)siRNA转染人脐静脉内皮EAhy926细胞后CSE的表达及对H2S的影响。方法体外培养EAhy926细胞,经siRNA转染以沉默CSE基因表达,分为5组进行培养:正常对照组(A组):只加入转染试剂;阴性对照组(B组):转染Neg-siRNA;转染CSEsiRNA1组(C组);转染CSEsiRNA2组(D组);转染CSEsiRNA3组(E组)。采用Lipofectamine 2000介导转染EAhy926细胞,在荧光显微镜下观察细胞以检测转染效率;Western blot法检测CSE蛋白表达;收集细胞检测H2S含量。结果观察转染效率为60%左右;A组CSE蛋白表达为(153.41±23.43),B组24 h CSE蛋白表达为(150.22±17.56),而D组24 h CSE蛋白表达降至(59.43±21.81),差异有统计学意义(P<0.01);并且A组H2S含量为(0.96±0.05),B组24 h H2S含量为(0.95±0.06),D组24 h H2S含量降至(0.54±0.08),差异有统计学意义(P<0.01)。结论 CSE-siRNA能够有效地抑制EAhy926中CSE的表达,并且CSE-siRNA2沉默作用最强。Objective To investigate the expression of cystathionine gamma lyase（CSE） after transfecting CSE small interfering RNA in human umbilical vein endothelial cell EAhy926 and the effect on H 2 S content.Methods EAhy926 cells were cultured in vitro,and were transfected by siRNA to knock down the expression of CSE.EAhy926 cells were divided into five groups： normal group（group A） ： only to add the transfection reagents;negative control group（group B） ： transfecting Neg-siRNA;transfecting CSEsiRNA1（group C）;transfecting CSEsiRNA2（group D）;transfecting CSEsiRNA3（group E）.Transfecting EAhy926 cells with Lipofectamine 2000,detected the transfecting efficiency under flourence microscope,tested CSE protein expression by Western blot,collected the cells to test the content of H 2 S.Results The transfection efficiency was about 60%.The expression of CSE protein in group A was 153.41 ± 23.43 and 150.22 ± 17.56 in group B after 24 h,whereras decreased to 59.43 ± 21.81 in group D after 24 h,there was statistic difference among them（P 〈 0.01）;and the content of H 2 S was 0.96 ± 0.05 in group A and 0.95 ± 0.06 in group B after 24 h,however decreased to 0.54 ± 0.08 in group D after 24 h,also had statistic difference among them（P 〈 0.01）.Conclusion CSEsiRNA can inhibit the expression of CSE in the EAhy926 cells effectively,and CSEsiRNA2 shows the strongest gene silence effect.

关 键 词：胱硫醚γ裂解酶 硫化氢 EAhy926 小干扰RNA 

分 类 号：R194.3[医药卫生—卫生事业管理] R977.4[医药卫生—公共卫生与预防医学]