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...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。展开更多
Background Hydrogen sulfide （H2S） plays an important role in the smooth muscle cell relaxation and thereby participates in the development of hypertension. Cystathionine γ-lyase is the key enzyme in the endogenous ...Background Hydrogen sulfide （H2S） plays an important role in the smooth muscle cell relaxation and thereby participates in the development of hypertension. Cystathionine γ-lyase is the key enzyme in the endogenous production of H2S. Up to now, the reports on the relationship between the polymorphisms of cystathionine γ-lyase gene （CTH） and essential hypertension （EH） are limited. This study was designed to assess their underlying relationship. Methods A total of 503 hypertensive patients and 490 age-, gender- and area-matched normotensive controls were enrolled in this study. Based on the FASTSNP, a web server to identify putative functional single nucleotide polymorphisms （SNPs） of genes, we selected two SNPs, rs482843 and rs1021737, in the CTH gene for genotyping. Genotyping was performed by the polymerase chain reaction and restriction fragment length polymorphism method （PCR-RFLP）. The frequencies of the alleles and genotypes between cases and controls were compared by the chi-square test. The program Haplo.stats was used to investigate the relationship between the haplotypes and EH. Results These two SNPs were in Hardy-Weinberg Equilibrium in both cases and controls. The genotype distribution and allele frequencies of them did not significantly differ between cases and controls （all P〉0.05）. In the stepwise logistic regression analysis we failed to observe their association with hypertension. In addition, none of the four estimated haplotypes or diplotypes significantly increased or decreased the risk of hypertension before or after adjustment for several known risk factors. Conclusions The present study suggests that the SNPs rs482843 and rs1021737 of the CTH gene were not associated with essential hypertension in the Northern Chinese Han population. However, replications in other populations and further functional studies are still necessary to clarify the role of the CTH gene in the pathogenesis of EH.展开更多
目的探讨是否可通过使用钙敏感受体(CaSR)激动剂增强胱硫醚-γ-裂解酶(CSE)的表达促使内源性硫化氢(H_2S)的含量增加,从而抑制肺动脉平滑肌细胞(PASMC)的增殖、迁移。方法以人肺动脉平滑肌细胞(HPASMC)为研究对象,将样本设置为空白对照...目的探讨是否可通过使用钙敏感受体(CaSR)激动剂增强胱硫醚-γ-裂解酶(CSE)的表达促使内源性硫化氢(H_2S)的含量增加,从而抑制肺动脉平滑肌细胞(PASMC)的增殖、迁移。方法以人肺动脉平滑肌细胞(HPASMC)为研究对象,将样本设置为空白对照组(使用PBS液干预)、GdCl_3组(使用CaSR激动剂GdCl_3干预)、NPS2390组(使用CaSR抑制剂NPS2390干预),在样本中加药孵育后,检测比较各组细胞增殖、迁移情况和细胞中Ca SR、CSE、H_2S的水平。结果三组细胞增殖、迁移情况和细胞中CaSR、CSE、H_2S的水平比较差异有统计学意义(P<0.05)。GdCl_3组细胞CaSR、CSE的表达和H2S的含量高于对照组,MTT实验吸光度(OD值)低于对照组,细胞迁移数少于对照组,差异有统计学意义(P<0.05);NPS2390组细胞CaSR的表达和H2S的水平均低于对照组,MTT实验吸光度(OD值)高于对照组,细胞迁移数多于对照组,差异有统计学意义(P<0.05)。结论 Ca SR激动剂可通过增强CSE的表达增加H_2S的含量,进而抑制PASMC的增殖及迁移。展开更多
文摘目的观察胱硫醚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。
基金This work was supported by the grants of the National Basic Research Program of China （No. 2006CB503805） and the Beijing Natural Science Foundation （No. 7061006）.
文摘Background Hydrogen sulfide （H2S） plays an important role in the smooth muscle cell relaxation and thereby participates in the development of hypertension. Cystathionine γ-lyase is the key enzyme in the endogenous production of H2S. Up to now, the reports on the relationship between the polymorphisms of cystathionine γ-lyase gene （CTH） and essential hypertension （EH） are limited. This study was designed to assess their underlying relationship. Methods A total of 503 hypertensive patients and 490 age-, gender- and area-matched normotensive controls were enrolled in this study. Based on the FASTSNP, a web server to identify putative functional single nucleotide polymorphisms （SNPs） of genes, we selected two SNPs, rs482843 and rs1021737, in the CTH gene for genotyping. Genotyping was performed by the polymerase chain reaction and restriction fragment length polymorphism method （PCR-RFLP）. The frequencies of the alleles and genotypes between cases and controls were compared by the chi-square test. The program Haplo.stats was used to investigate the relationship between the haplotypes and EH. Results These two SNPs were in Hardy-Weinberg Equilibrium in both cases and controls. The genotype distribution and allele frequencies of them did not significantly differ between cases and controls （all P〉0.05）. In the stepwise logistic regression analysis we failed to observe their association with hypertension. In addition, none of the four estimated haplotypes or diplotypes significantly increased or decreased the risk of hypertension before or after adjustment for several known risk factors. Conclusions The present study suggests that the SNPs rs482843 and rs1021737 of the CTH gene were not associated with essential hypertension in the Northern Chinese Han population. However, replications in other populations and further functional studies are still necessary to clarify the role of the CTH gene in the pathogenesis of EH.
文摘目的探讨是否可通过使用钙敏感受体(CaSR)激动剂增强胱硫醚-γ-裂解酶(CSE)的表达促使内源性硫化氢(H_2S)的含量增加,从而抑制肺动脉平滑肌细胞(PASMC)的增殖、迁移。方法以人肺动脉平滑肌细胞(HPASMC)为研究对象,将样本设置为空白对照组(使用PBS液干预)、GdCl_3组(使用CaSR激动剂GdCl_3干预)、NPS2390组(使用CaSR抑制剂NPS2390干预),在样本中加药孵育后,检测比较各组细胞增殖、迁移情况和细胞中Ca SR、CSE、H_2S的水平。结果三组细胞增殖、迁移情况和细胞中CaSR、CSE、H_2S的水平比较差异有统计学意义(P<0.05)。GdCl_3组细胞CaSR、CSE的表达和H2S的含量高于对照组,MTT实验吸光度(OD值)低于对照组,细胞迁移数少于对照组,差异有统计学意义(P<0.05);NPS2390组细胞CaSR的表达和H2S的水平均低于对照组,MTT实验吸光度(OD值)高于对照组,细胞迁移数多于对照组,差异有统计学意义(P<0.05)。结论 Ca SR激动剂可通过增强CSE的表达增加H_2S的含量,进而抑制PASMC的增殖及迁移。
