A study on the effect of static stretch on the expression of NOS in glomerular endothelial cells

静态拉伸对肾小球内皮细胞NOS表达影响的研究

• 批准号: 09671180
• 负责人: KAWAMURA Tetsuya
• 金额: $ 1.66万
• 依托单位: Jikei University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671180/
• 关键词: glomerular endothelial cell; static stretch; mecharical strain; nitric oxide; endothelial NOS; vascular endothelial cell; cyclic GMP; brady kinin; MCP-1; 糸球体硬化; マクロファージ; メサンギウム細胞; 細胞外基質; ICAM-1; M-CSF; glomerular endothelial cell; static stretch; mecharical strain; nitric oxide; endothelial NOS; vascular endothelial cell; cyclic GMP; brady kinin; MCP-1; 糸球体硬化; マクロファージ; メサンギウム細胞; 細胞外基質; ICAM-1; M-CSF

In the present study, we examined the effect of static stretch to glomerular endothelial cells (GEC) and vascular endothelial cells (VEC) on the expression of nitric oxide synthase (NOS) and the production of nitric oxide (NO) by these cells upon the stimulation by bradykinin.Northern blot analysis and RT-PCR revealed that mRNA levels for endothelial NOS (eNOS) were not significantly different between the cells with and without exposure to 30-40 % static stretch for up to 48hr. In contrast, it was clearly demonstrated by Western blot analysis that the expression of eNOS protein was down-regulated in VEC exposed to static stretch for 24 hours in a stretch-degree dependent manner. Moreover, 10^<-6> M bradykinin markedly increased cGMP contents in untreated VEC, whereas this effect was completely abolished when VEC were exposed to 30% stretch for 24 hours prior to the stimulation by bradykinin. These results indicate that the expression of eNOS protein may be down-regulated in VEC when they are exposed to a high degree of mechanical stretch, leading to a compromised production of NO in response to eNOS agonists. We are in the process of examining the effect of static stretch to GEC on the production of NO upon the stimulation by bradykinin, in order to elucidate a role of a high degree of mechanical stretch to GEC in the regulation of glomerular microcirculation.

In the present study, we examined the effect of static stretch to glomerular endothelial cells (GEC) and vascular endothelial cells (VEC) on the expression of nitric oxide synthase (NOS) and the production of nitric oxide (NO) by these cells upon the stimulation by bradykinin.Northern blot analysis and RT-PCR revealed that mRNA levels for endothelial NOS (eNOS) were not significantly different between the有或没有暴露于30-40％静态伸展的细胞可容纳48小时。相比之下，通过蛋白质印迹分析清楚地证明了eNOS蛋白的表达在VEC中下调，以静态延伸的静态伸展24小时，以伸展度的依赖性方式下调。此外，在未处理的VEC中，10^<-6> mbradykinin显着增加了CGMP含量，而当VEC暴露于Bradykinin刺激前24小时之前，该效果完全消除了30％的延伸。这些结果表明，当ENOS蛋白暴露于高度机械伸展时，ENOS蛋白的表达可能会下调，从而导致对eNOS激动剂的响应，导致NO的产生受损。我们正在研究静态拉伸对GEC对Bradykinin刺激的产生的影响，以阐明高度机械伸展对GEC在肾小球微循环调节中的作用。