高盐培养基对人脐静脉内皮细胞株eNOS表达的影响及其机制

发布时间：2018-10-11 11:09

【摘要】：目的探讨高盐培养基干预人脐静脉细胞融合细胞(EA.hy926)对eNOS表达的影响及相关机制。方法予以EA.hy926细胞不同浓度的高盐培养基干预48h,选择137mmol/L氯化钠为高盐培养基的浓度;分别予以DMEM高糖培养基组(氯化钠浓度为109mmol/L)、高盐培养基(氯化钠浓度为137mmol/L)、高盐培养基(氯化钠浓度为137mmol/L)+RNA干扰、高渗对照组(甘露醇浓度为56mmol/L)干预48h,高效液相色谱法检测细胞上清液非对称二甲基精氨酸(ADMA)浓度,实时定量PCR和Western blot检测蛋白质精氨酸甲基转移酶1(PRMT-1)、RhoA、Rho激酶(ROCK)、内皮型一氧化氮合酶(eNOS)的表达及活性。结果高盐组上清液中ADMA浓度、PRMT-1表达、RhoA mRNA表达及ROCK活性较对照组显著增高,eNOS蛋白表达显著降低;高盐培养基+RNA干扰组较高盐组ADMA浓度、PRMT-1、RhoA mRNA及ROCK活性明显下降,eNOS蛋白表达显著上调。结论高盐培养基通过刺激ADMA生成并激活下游RhoA-ROCK通路而抑制eNOS的生成。
[Abstract]:Objective to investigate the effect of high salt medium on the expression of eNOS in human umbilical vein cell fusion cells (EA.hy926). Methods EA.hy926 cells were treated with different concentrations of high salt medium for 48 h, and 137mmol/L sodium chloride was selected as the concentration of high salt medium. DMEM high glucose medium group (sodium chloride concentration 109mmol/L), high salt medium (sodium chloride concentration 137mmol/L), high salt medium (sodium chloride concentration 137mmol/L) RNA interference were given, respectively. The concentration of asymmetric dimethyl arginine (ADMA) in the supernatant of cells was detected by HPLC for 48 h in hypertonic control group (mannitol concentration: 56mmol/L). The expression and activity of protein arginine methyltransferase 1 (PRMT-1) and RhoA,Rho kinase (ROCK), endothelial nitric oxide synthase (eNOS) were detected by real-time quantitative PCR and Western blot. Results the concentration of ADMA, the expression of, RhoA mRNA and the activity of ROCK in the supernatant of high salt group were significantly higher than those in the control group, and the expression of eNOS protein was significantly lower than that in the control group. The concentration of ADMA, the activities of PRMT-1,RhoA mRNA and ROCK in the RNA interference group were significantly decreased, and the expression of eNOS protein was upregulated. Conclusion High salt medium inhibits eNOS production by stimulating ADMA production and activating downstream RhoA-ROCK pathway.
【作者单位】：西安交通大学医学院第一附属医院心内科;北方民族大学信息与计算科学学院;
【基金】：国家自然科学基金资助项目(No.30900616) 国家重点基础研究发展规划(973计划,No.2012CB517804)~~
【分类号】：R363

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