miRNA-200a对人主动脉内皮细胞间质转化的影响及其机制

doi:10.3969/j.issn.1671-7171.2017.09.008

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摘要【目的】探索miR-200a靶向抑制高迁移率族蛋白B1 (high-mobility group protein box 1, HMGB1)调控转化生长因子-β1(transforming growth factor-β1, TGF-β1)诱导的人主动脉内皮细胞间质转化(endothelial-mesenchymal transition, EndMT)。【方法】①不同浓度TGF-β1处理后miR-200a的表达差异。②miR-200a过表达后，经或不经TGF-β1处理的HAEC中内皮间质转化标记因子血小板内皮细胞黏附分子（platelet endothelial cell adhesion molecule-1, CD31）、血管内皮钙粘素（vascular endothelial cadherin, VE-cadherin）、成纤维细胞特异性蛋白（fibroblast-specific protein-1, FSP-1）和α-平滑肌肌动蛋白（α-smooth muscle actin, α-SMA）的表达差异。③miR-200a过表达和TGF-20处理对HMGB1的表达差异。④荧光素酶实验检测miR-200a与HMGB1 3’UTR的靶向结合关系。⑤miR-200a mimics和HMGB1单独或联合转染的HAEC中CD31、VE-cadherin、FSP-1及α-SMA的蛋白表达差异。【结果】①TGF-β1可抑制miR-200a表达(P<0.01)。②miR-200a抑制TGF-β1诱导的EndMT过程，上调CD31(P<0.01)、VE-cadherin (P<0.001)，下调FSP-1(P<0.001)、α-SMA(P<0.01)。③miR-200a抑制HMGB1的表达(P<0.001)，TGF-00促进HMGB1的表达(P<0.001)。④miR-200a与HMGB1的3’UTR区域存在靶向结合。⑤HMGB1减弱miR-200a的抑制EndMT过程，下调CD31(P<0.01)、VE-cadherin(P<0.01)，上调α-SMA (P<0.01)、FSP-1(P<0.005)。【结论】HMGB1促进人主动脉EndMT，miR-200a抑制HMGB1调控TGF-β1诱导的人主动脉EndMT。

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	熊盈
	孙艳
	李英
	但平

关键词 ：微RNAs, 主动脉, 内皮细胞/细胞学, 生物转化, 转化生长因子β1, 高迁移率族蛋白质类

Abstract：【Objective】To investigate the effect and the underlying mechanism of miRNA(miR)-200a in regulating TGF-β1-induced endothelial-mesenchymal transition (EndMT) of human aortic endothelial cells (HAVC) by targeting inhibition of high-mobility group protein box1, (HMGB1) .【Methods】1. The expression of miR-200a in human aortic endothelial cell lysates (HAEC) was detected under different concentrations of TGF-β1. 2. The expressions of platelet endothelial cell adhesion molecule-1 (CD31), endothelium-cadherin (VE-cadherin), fibroblast-specific protein-1 (FSP-1) andα-smooth muscle actin (α-SMA) were determined after miR-200a mimics/mimic NC transfection occurred in HAEC with or without TGF-β1 treatment. 3. The expression levels of HMGB1 in miRNA-200a mimics/mimic NC-transfected HAECs that were and were not treated with TGF-β1 were determined. 4. The binding relationship between miR-200a and HMGB1 3'UTR was detected by luciferase assay. 5. The protein expressions of CD31, VE-cadherin, FSP-1 and α-SMA in HAEC transfected with miR-200a mimics and HMGB1 alone or in combination were detected.【Results】1. TGF-β1 inhibited the expression of miR-200a(P＜0.01). 2. miR-200a inhibited TGF-β1-induced EndMT, up-regulated CD31(P＜0.01) and VE-cadherin(P＜0.001), while down- regulating FSP-1 (P＜0.001) and α-SMA (P＜0.01) expression. 3. miR-200a inhibited the expression of HMGB1(P＜0.001), while TGF- β1 promoted the expression of HMGB1(P＜0.001). 4. miR-200a binded to the 3'UTR region of HMGB1. 5. HMGB1 attenuated miR-200a inhibition of EndMT and down-regulated the expression of CD31(P＜0.01) and VE-cadherin(P＜0.01), while up-regulated the expression of α-SMA(P＜0.01) and FSP-1(P＜0.001).【Conclusion】The transfection of HMGB1 plasmid promoted the EndMT process of human aortic endothelial cell.The transfection of miR-200a inhibited the expression of HMGB1 and inhibited EndMT of human aortic endothelial cell induced by TGF-β1.Therefore miRNA(miR)-200a targets the inhibition of high-mobility group protein box1, (HMGB1) in regulating TGF-β1-induced endothelial-mesenchymal transition.

Key words： MicroRNAs Aorta Endothelial Cells/CY Biotransformation Transforming Growth Factor beta1 High Mobility Group Proteins

收稿日期: 2017-05-22

PACS:

R543.1

引用本文:

熊盈,孙艳,李英,但平. miRNA-200a对人主动脉内皮细胞间质转化的影响及其机制[J]. 医学临床研究, 2017, 34(9): 1691-1696.
XIONG Ying, SUN Yan, LI Ying,et al. Effect and the Underlying Mechanism of miRNA-200a in Regulating TGF-β1-induced Endothelial-mesenchymal Transition of Human Aortic Endothelial Cells. JOURNAL OF CLINICAL RESEARCH, 2017, 34(9): 1691-1696.

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http://www.jcr.net.cn/CN/10.3969/j.issn.1671-7171.2017.09.008 或 http://www.jcr.net.cn/CN/Y2017/V34/I9/1691

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