Diabetic vascular complications caused by endothelial dysfunction play an important role in the pathogenesis of diabetic foot. A well understanding of the role of endothelial dysfunction in diabetic foot vasculopathy will help to further reveal the pathogenesis of diabetic foot. This study aimed to assess whether the RhoA/ROCK signaling pathway is controlled by Rho GTPase-activating proteins (RhoGAP, ARHGAP) and advanced glycosylation end products (AGEs), and to clarify the roles of ARHGAP and AGEs in the RhoA/ROCK signaling pathway or the mechanism by which AGEs regulated RhoA. Real-time PCR was applied to detect gene expression. Manipulation of endothelial biological functions by ARHGAP18 and AGEs were studied via cell counting kit-8 (CCK-8), Western blot, transwell, FITC-Dextran and TEER permeability experiments. RhoA-specific inhibitor Y-27632 was used to silence the activity of RhoA. Dual Luciferase Reporter Assay, Western blot and ELISA assays were used to detect molecular mechanism of endothelial biological functions. In this study, we found that ARHGAP18 was negatively correlated with RhoA, and the expression of ARHGAP18 in human umbilical vein endothelial cells (HUVECs) was decreased with gradient-increased AGEs. Furthermore, AGEs and ARHGAP18 could orchestrate RhoA activity, then activate NF-κB signaling pathway, affect the structural and morphological of VE-cadherin and tight junction protein, and cause endothelial cell contraction, thereby increasing permeability of endothelial cells. In conclusion, AGEs and ARHGAP18 orchestrate cell proliferation, invasion and permeability by controlling the RhoA/ROCK signaling pathway, affecting NF-κB signaling pathway as well as the structure and morphology of VE-cadherin and tight junction protein, and regulating endothelial cell contraction.

中文翻译：

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先进的糖基化终产物（AGEs）通过协调人脐静脉内皮细胞中的ARHGAP18 / RhoA途径来控制增殖，侵袭和渗透性。

内皮功能障碍引起的糖尿病血管并发症在糖尿病足的发病机理中起重要作用。充分了解内皮功能障碍在糖尿病足血管病变中的作用将有助于进一步揭示糖尿病足的发病机理。这项研究旨在评估RhoA / ROCK信号通路是否受Rho GTPase激活蛋白（RhoGAP，ARHGAP）和高级糖基化终产物（AGEs）的控制，并阐明ARHGAP和AGEs在RhoA / ROCK信号通路中的作用或AGEs调节RhoA的机制。应用实时PCR检测基因表达。通过细胞计数试剂盒8（CCK-8），蛋白质印迹，transwell，FITC-Dextran和TEER渗透性实验研究了ARHGAP18和AGEs对内皮生物学功能的操纵。RhoA特异性抑制剂Y-27632用于沉默RhoA的活性。采用双重荧光素酶报告基因测定，Western印迹和ELISA测定来检测内皮生物学功能的分子机制。在这项研究中，我们发现ARHGAP18与RhoA呈负相关，并且随着年龄的增加AGEs，ARHGAP18在人脐静脉内皮细胞（HUVECs）中的表达降低。此外，AGEs和ARHGAP18可以协调RhoA活性，然后激活NF-κB信号通路，影响VE-钙粘蛋白和紧密连接蛋白的结构和形态，并引起内皮细胞收缩，从而增加内皮细胞的通透性。总之，AGEs和ARHGAP18通过控制RhoA / ROCK信号传导途径来协调细胞的增殖，侵袭和通透性，