黄大祥,李梦媛,秦 岭,彭 艾.RhoA/ROCK/ERK-MAPK通路在高磷环境调节内皮细胞凋亡的机制[J].转化医学杂志,2021,10(4):224-227
RhoA/ROCK/ERK-MAPK通路在高磷环境调节内皮细胞凋亡的机制
A Study on the mechanism of high phosphorus regulating endothelial cell apoptosis through RhoA/ROCK/ERK-MAPK pathway
  
DOI:
中文关键词:  高磷  Ras同源物基因家族成员A  磷酸化细胞外信号调控激酶  精氨酸酶  内皮细胞
英文关键词:High phosphorus  Ras homolog gene family, member A, (RhoA)  Extracellular signal-regulated kinase(ERK)  Arginase(ARG)  Endothelial cells
基金项目:上海市崇明区科学技术委员会“可持续发展科技创新行动计划”(CKY2018-24);上海市第十人民医院国自然培育项目-面上A类 (04.03.19.101)
作者单位
黄大祥 上海市第十人民医院崇明分院肾脏科 
李梦媛 同济大学附属第十人民医院肾脏科 
秦 岭 同济大学附属第十人民医院肾脏科 
彭 艾 同济大学附属第十人民医院肾脏科 
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中文摘要:
      目的 研究高磷环境下内皮细胞凋亡的可能调控机制和信号通路。方法 人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)在正常磷(1.0 mmol/L)、高磷(3.0 mmol/L)、正常磷+辛伐他汀(simvastatin,SV,1.0 mmol /L+SV)及高磷+SV (3.0 mmol /L+SV)培养基中培养24 h,Western Blot评价Ras同源物基因家族成员A( Ras homolog gene family, member A, RhoA),Rho相关卷曲螺旋形成蛋白激酶1/2 ( rho-associated coiled coil forming protein kinase 1/2, ROCK1/2 ),细胞外信号调控激酶-丝裂原活化蛋白激酶(extracellular signal-regulated kinase-mitogen activated protein kinase, ERK-MAPK)、磷酸化细胞外信号调控激酶-丝裂原活化蛋白激酶( phosphorylated-ERK-MAPK, p-ERK-MAPK)、p38丝裂原活化蛋白激酶 ( p38-Mitogen Activated Protein Kinase , p38-MAPK)、磷酸化 p38-丝裂原活化蛋白激酶 (phosphorylated-p38-MAPK, p-p38-MAPK )、以及精氨酸酶1(arginase1,ARG1)和一氧化氮合成酶(nitric oxide synthase, NOS )蛋白的表达,比色法和膜联蛋白V/碘化丙啶双染法 ( Annexin V-FITC/ propidium iodide, Annexin V-FITC/PI ) 检测ARG活性和细胞凋亡率。结果 高磷组RhoA,ROCK1/2,p-ERK MAPK,ARG1表达均上调, ARG活性升高,NOS表达下调,细胞凋亡增加;RhoA抑制剂SV能抑制高磷环境下p-ERK MAPK的活化和ARG1 的表达,降低ARG1活性,促进NOS表达,减少内皮细胞凋亡。结论 高磷环境HUVECs可通过RhoA/ROCK途径激活p-ERK MAPK的表达, 增加ARG活性,抑制NOS表达,引起内皮细胞凋亡;SV可以通过抑制RhoA/ROCK/ERK-MAPK通路,改善内皮细胞凋亡。
英文摘要:
      Objective To study the exact mechanism of high phosphorus on endothelial cell apoptosis and functional disorders. Methods Human umbilical vein endothelial cells (HUVECs) were cultured and divided into normal phosphorus concentration (1.0 mmol/L) , high phosphorus concentration ( 3.0 mmol/L ),normal phosphorus plus simvastatin(1.0 mmol /L+SV)and high phosphorus plus simvastatin (3.0 mmol /L+SV). Western blot was used to evaluate Ras homolog gene family, member A ( RhoA ) , Rho-associated Coiled Coil Forming Protein Kinase 1/2 (ROCK1/2), Extracellular signal-Regulated Kinase-Mitogen Activated Protein Kinase (ERK- MAPK), phosphorylated-ERK-MAPK(p-ERK-MAPK), p38-Mitogen Activated Protein Kinase(p38-MAPK, phosphorylated-p38-Mitogen Activated Protein Kinase(p-p38-MAPK), arginase (Arginase1, ARG1) and nitric oxide synthase (NOS) protein expression. Colorimetry was used to assay the activity of arginase. Annexin V-FITC /propidium iodide(AnnexinV-FITC/PI)double staining was used to assess the apoptosis of cells. Results RhoA, ROCK1/2, p-ERK MAPK, and ARG1 protein was up-regulated, and the expression of NOS was down-regulated in the high phosphorus group; the RhoA inhibitor simvastatin can inhibit the activation of p-ERK MAPK and the expression of ARG1, promote the expression of NOS and decreased the endothelial cells apoptosis of the high phosphorus group. Conclusion HUVECs may activate the expression of p-ERK MAPK through the RhoA/ROCK pathway to regulate arginase activity under high phosphorus environment, thereby inhibiting the activity of NOS, causing endothelial cell apoptosis. Simvastatin can decrease the apoptosis of endothelial cells by inhibiting RhoA/ROCK/ERK-MAPK pathway.
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