{"title":"YAP/TAZ信号在高糖环境中促进视网膜微血管内皮细胞的血管生成","authors":"Xing-Li Wang, Yang Xian, Xiao-Long Chen","doi":"10.1080/02713683.2024.2309217","DOIUrl":null,"url":null,"abstract":"<p><strong>Purpose: </strong>Diabetic retinopathy (DR) is a major cause of irreversible blindness in the working-age population. Neovascularization is an important hallmark of advanced DR. There is evidence that Yes-associated protein (YAP)/transcriptional co-activator with a PDZ binding domain (TAZ) plays an important role in angiogenesis and that its activity is regulated by vascular endothelial growth factor (VEGF). Therefore, the aim of this study was to investigate the effect of YAP/TAZ-VEGF crosstalk on the angiogenic capacity of human retinal microvascular endothelial cells (hRECs) in a high-glucose environment.</p><p><strong>Methods: </strong>The expression of YAP and TAZ of hRECs under normal conditions, hypertonic conditions and high glucose were observed. YAP overexpression (OE-YAP), YAP silencing (sh-YAP), VEGF overexpression (OE-VEGF) and VEGF silencing (sh-VEGF) plasmids were constructed. Cell counting kit-8 assay was performed to detect cells proliferation ability, transwell assay to detect cells migration ability, and tube formation assay to detect tube formation ability. The protein expression of YAP, TAZ, VEGF, matrix metalloproteinase (MMP)-8, MMP-13, vessel endothelium (VE)-cadherin and alpha smooth muscle actin (α-SMA) was measured by western blot.</p><p><strong>Results: </strong>The proliferation of hRECs was significantly higher in the high glucose group compared with the normal group, as well as the protein expression of YAP and TAZ (<i>p</i> < 0.01). YAP and VEGF promoted the proliferation, migration and tube formation of hRECs in the high glucose environment (<i>p</i> < 0.01), and increased the expression of TAZ, VEGF, MMP-8, MMP-13 and α-SMA while reducing the expression of VE-cadherin (<i>p</i> < 0.01). Knockdown of YAP effectively reversed the above promoting effects of OE-VEGF (<i>p</i> < 0.01) and overexpression of YAP significantly reversed the inhibition effects of sh-VEGF on above cell function (<i>p</i> < 0.01).</p><p><strong>Conclusion: </strong>In a high-glucose environment, YAP/TAZ can significantly promote the proliferation, migration and tube formation ability of hRECs, and the mechanism may be related to the regulation of VEGF expression.</p>","PeriodicalId":10782,"journal":{"name":"Current Eye Research","volume":null,"pages":null},"PeriodicalIF":1.7000,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"YAP/TAZ Signaling Enhances Angiogenesis of Retinal Microvascular Endothelial Cells in a High-Glucose Environment.\",\"authors\":\"Xing-Li Wang, Yang Xian, Xiao-Long Chen\",\"doi\":\"10.1080/02713683.2024.2309217\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Purpose: </strong>Diabetic retinopathy (DR) is a major cause of irreversible blindness in the working-age population. Neovascularization is an important hallmark of advanced DR. There is evidence that Yes-associated protein (YAP)/transcriptional co-activator with a PDZ binding domain (TAZ) plays an important role in angiogenesis and that its activity is regulated by vascular endothelial growth factor (VEGF). Therefore, the aim of this study was to investigate the effect of YAP/TAZ-VEGF crosstalk on the angiogenic capacity of human retinal microvascular endothelial cells (hRECs) in a high-glucose environment.</p><p><strong>Methods: </strong>The expression of YAP and TAZ of hRECs under normal conditions, hypertonic conditions and high glucose were observed. YAP overexpression (OE-YAP), YAP silencing (sh-YAP), VEGF overexpression (OE-VEGF) and VEGF silencing (sh-VEGF) plasmids were constructed. Cell counting kit-8 assay was performed to detect cells proliferation ability, transwell assay to detect cells migration ability, and tube formation assay to detect tube formation ability. The protein expression of YAP, TAZ, VEGF, matrix metalloproteinase (MMP)-8, MMP-13, vessel endothelium (VE)-cadherin and alpha smooth muscle actin (α-SMA) was measured by western blot.</p><p><strong>Results: </strong>The proliferation of hRECs was significantly higher in the high glucose group compared with the normal group, as well as the protein expression of YAP and TAZ (<i>p</i> < 0.01). YAP and VEGF promoted the proliferation, migration and tube formation of hRECs in the high glucose environment (<i>p</i> < 0.01), and increased the expression of TAZ, VEGF, MMP-8, MMP-13 and α-SMA while reducing the expression of VE-cadherin (<i>p</i> < 0.01). Knockdown of YAP effectively reversed the above promoting effects of OE-VEGF (<i>p</i> < 0.01) and overexpression of YAP significantly reversed the inhibition effects of sh-VEGF on above cell function (<i>p</i> < 0.01).</p><p><strong>Conclusion: </strong>In a high-glucose environment, YAP/TAZ can significantly promote the proliferation, migration and tube formation ability of hRECs, and the mechanism may be related to the regulation of VEGF expression.</p>\",\"PeriodicalId\":10782,\"journal\":{\"name\":\"Current Eye Research\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.7000,\"publicationDate\":\"2024-05-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Eye Research\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1080/02713683.2024.2309217\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/2/2 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q3\",\"JCRName\":\"OPHTHALMOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Eye Research","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1080/02713683.2024.2309217","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/2/2 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"OPHTHALMOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
目的:糖尿病视网膜病变(DR)是导致劳动适龄人口不可逆转性失明的主要原因。血管新生是晚期糖尿病视网膜病变的重要标志。有证据表明,Yes 相关蛋白(YAP)/具有 PDZ 结合域的转录共激活因子(TAZ)在血管生成中发挥着重要作用,其活性受血管内皮生长因子(VEGF)的调控。因此,本研究旨在探讨YAP/TAZ-VEGF串扰对高糖环境下人视网膜微血管内皮细胞(hRECs)血管生成能力的影响:方法:观察正常、高渗和高糖条件下 hRECs 的 YAP 和 TAZ 表达。构建了YAP过表达(OE-YAP)、YAP沉默(sh-YAP)、VEGF过表达(OE-VEGF)和VEGF沉默(sh-VEGF)质粒。细胞计数试剂盒-8 检测细胞增殖能力,Transwell 检测细胞迁移能力,试管形成检测试管形成能力。通过 Western blot 检测 YAP、TAZ、VEGF、基质金属蛋白酶(MMP)-8、MMP-13、血管内皮(VE)-cadherin 和α-平滑肌肌动蛋白(α-SMA)的蛋白表达:结果:与正常组相比,高糖组 hRECs 的增殖明显增加,YAP 和 TAZ 的蛋白表达也明显增加(p p p p p 结论):在高糖环境中,YAP/TAZ能明显促进hRECs的增殖、迁移和管形成能力,其机制可能与VEGF的表达调控有关。
YAP/TAZ Signaling Enhances Angiogenesis of Retinal Microvascular Endothelial Cells in a High-Glucose Environment.
Purpose: Diabetic retinopathy (DR) is a major cause of irreversible blindness in the working-age population. Neovascularization is an important hallmark of advanced DR. There is evidence that Yes-associated protein (YAP)/transcriptional co-activator with a PDZ binding domain (TAZ) plays an important role in angiogenesis and that its activity is regulated by vascular endothelial growth factor (VEGF). Therefore, the aim of this study was to investigate the effect of YAP/TAZ-VEGF crosstalk on the angiogenic capacity of human retinal microvascular endothelial cells (hRECs) in a high-glucose environment.
Methods: The expression of YAP and TAZ of hRECs under normal conditions, hypertonic conditions and high glucose were observed. YAP overexpression (OE-YAP), YAP silencing (sh-YAP), VEGF overexpression (OE-VEGF) and VEGF silencing (sh-VEGF) plasmids were constructed. Cell counting kit-8 assay was performed to detect cells proliferation ability, transwell assay to detect cells migration ability, and tube formation assay to detect tube formation ability. The protein expression of YAP, TAZ, VEGF, matrix metalloproteinase (MMP)-8, MMP-13, vessel endothelium (VE)-cadherin and alpha smooth muscle actin (α-SMA) was measured by western blot.
Results: The proliferation of hRECs was significantly higher in the high glucose group compared with the normal group, as well as the protein expression of YAP and TAZ (p < 0.01). YAP and VEGF promoted the proliferation, migration and tube formation of hRECs in the high glucose environment (p < 0.01), and increased the expression of TAZ, VEGF, MMP-8, MMP-13 and α-SMA while reducing the expression of VE-cadherin (p < 0.01). Knockdown of YAP effectively reversed the above promoting effects of OE-VEGF (p < 0.01) and overexpression of YAP significantly reversed the inhibition effects of sh-VEGF on above cell function (p < 0.01).
Conclusion: In a high-glucose environment, YAP/TAZ can significantly promote the proliferation, migration and tube formation ability of hRECs, and the mechanism may be related to the regulation of VEGF expression.
期刊介绍:
The principal aim of Current Eye Research is to provide rapid publication of full papers, short communications and mini-reviews, all high quality. Current Eye Research publishes articles encompassing all the areas of eye research. Subject areas include the following: clinical research, anatomy, physiology, biophysics, biochemistry, pharmacology, developmental biology, microbiology and immunology.