siRNA基因沉默ASIC1a对过敏性紫癜(HSP)患儿血清IgA1诱导的血管内皮细胞损伤的保护作用

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英文篇名：Silencing of ASIC1a protects the vascular endothelial cells damage induced by IgA1 from Henoch-Schonlein syndrome patients 作者：闫波 ; 袁丽萍 ; 张琴 ; 华冉 ; 李玉飞 ; 戴寒晶 英文作者：YAN Bo;YUAN Li-ping;ZHANG Qin;HUA Ran;LI Yu-fei;DAI Han-jing;Department of medical technology,Anhui Medical College;Department of Pediatrics,First Affiliated Hospital of Anhui Medical University; 关键词：过敏性紫癜 ; 血管内皮细胞 ; ASIC1a 英文关键词：Henoch-Schonlein syndrome(HSP);;Vascular endothelial cells;;Acid-sensing ion channel1a(ASIC1a) 中文刊名：GNYX 英文刊名：Journal of Gannan Medical University 机构：安徽医学高等专科学校医学技术系;安徽医科大学第一附属医院儿科; 出版日期：2019-06-12 08:44 出版单位：赣南医学院学报 年：2019 期：v.39;No.177 基金：国家自然科学基金项目(81471617);; 安徽省高校自然科学重点科研项目(KJ2015A360) 语种：中文; 页：GNYX201904002 页数：6 CN：04 ISSN：36-1154/R 分类号：13-18

摘要

目的:探讨siRNA基因沉默ASIC1a对过敏性紫癜患儿血清Ig A1诱导的血管内皮细胞的保护作用。方法:体外培养人正常皮肤微血管内皮细胞(HDMVEC),设计针对人ASIC1a基因编码区的siRNA序列,构建重组慢病毒LV-sh-ASIC1a转染给HDMVEC细胞,设立空病毒转染对照组(NC组)、LV-h-ASIC1a转染组(si-ASIC1a组),采用RT-PCR检测转入基因ASIC1a的表达。病毒转染72 h后,加入分离的过敏性紫癜患儿血清Ig A1(HSP Ig A1)和正常患儿血清Ig A1,采用ELISA法检测细胞培养上清中TNF-α、IL-8水平,real-time PCR和Western blotting分别检测细胞内ASIC1a、细胞骨架蛋白(SM-α、Destrin、Vinculin、ML-CK) mRNA及蛋白表达水平。结果:与NC对照组比较,si-ASIC1a组HDMVEC内TNF-α、IL-8的分泌显著减少,细胞骨架蛋白SM-α、Destrin、Vinculin、ML-CK mRNA和蛋白表达明显增加,差异有统计学意义(P <0. 01)。结论:沉默ASIC1a可以显著抑制HSP血管内皮细胞细胞骨架蛋白的下调,改善血管内皮细胞损伤。
        Objective: To investigate the protective effect of silencing of ASIC1 a on the vascular endothelial cells damage induced by Ig A1 from Henoch-Schonlein syndrome( HSP) patients. Methods: Human dermal microvascular endothelial cells( HDMVEC) were cultured in vitro,siRNA sequences were designed for the coding region of human ASIC1 a gene and HDMVEC cells were transfected with recombinant lentivirus( LV)-sh-ASIC1 a. The control group( NC group) without virus transfection and LV-sh-ASIC1 a transfection group( si-ASIC1 a group) were set up. The expression of transfixed ASIC1 a gene was detected by RT-PCR. After virus transfection 72 h,serum Ig A1 from HSP patients and serum Ig A1 of normal children were added into HDMVEC cells. The levels of TNF-α and IL-8 in cell culture supernatant were detected by ELISA,ASIC1 a and cytoskeleton protein( SM-α,Destrin,Vinculin,ML-CK) mRNA and protein expressions were detected by real-time PCR and Western blotting Methods. Results: TNF-α、IL-8 release in the group of si-ASIC1 a group was significantly reduced and cytoskeleton protein( SM-α,Destrin,Vinculin,ML-CK) mRNA and protein expressions compared with the NC control group( P < 0. 01). Conclusions: Silencing ASIC1 a can protect the damage of HSP vascular endothelial cells.

引文

[1] Yu HH,Chiang BL,Yang YH. Altered glycosylation of circulatory Ig A1 involved in Henoch-Schnlein purpura and Ig A nephropathy[J]. J Formos Med Assoc,2012,111(3):121-122.
    [2]袁丽萍,张琴,鹿玲.过敏性紫癜患儿血管内皮细胞凋亡与血清Ig A水平关系探讨[J].中国免疫学杂志,2012,1:81-84.
    [3]闫波,袁丽萍,彭启迪,等.酸环境对过敏性紫癜患儿血清Ig A1诱导血管内皮细胞ASIC1a表达的影响及机制[J].基础医学与临床,2018,37(12):1674-1679.
    [4] Xu TL,Xiong ZG. Dynamic regulation of acid-sensing ion channels by extracellular and intracellular modulators[J].Curr Med Chem,2007,14(16):1753-1763.
    [5] Yang YH,Huang YH,Lin YL,et al. Circulating Ig A from acute stage of childhood Henoch-Schnlein purpura can enhance endothelial interleukin(IL)-8 production through MEK/ERK signalling pathway[J]. Clin Exp Immunol,2006,144(2):247-253.
    [6]张琴,鹿玲,林达.儿童过敏性紫癜血清Ig A1和皮肤小血管Ig A1的沉积及意义[J].中华皮肤病杂志,2008,41(1):55-56.
    [7] Xu H,Bi SY,Zheng ZQ,et al. Effects of omeprazole in the anaphylactoid purpura abdominalis of children[J].Chin J Gastroenterol Hepatol,2010,19:1025-1027.
    [8] MJ Dillon,Park SJ,Suh JS,et al. Advances in our understanding of the pathogenesis of Henoch-Schnlein purpura and the implications for improving its diagnosis[J].Expert Rev Clin Immunol,2013,9(12):1223-1238.
    [9] Alkjaer C,Peng HL. Effect of p H on vascular tension:which are the important mechanisms[J]. J Vasc Res,1997,161(4):557-566.