Chen Yanlan, Huang Wen, Jiang Wenlin, Wu Xianghong, Ye Biao, Zhou Xiaoting
Department of Neurology, First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China.
Department of Vasculocardiology, First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China.
Oxid Med Cell Longev. 2016;2016:4196572. doi: 10.1155/2016/4196572. Epub 2016 Aug 2.
HIV-1 transactivator protein (Tat) has been shown to play an important role in HIV-associated neurocognitive disorders. The aim of the present study was to evaluate the relationship between occludin and amyloid-beta (Aβ) transfer receptors in human cerebral microvascular endothelial cells (hCMEC/D3) in the context of HIV-1-related pathology. The protein expressions of occludin, receptor for advanced glycation end products (RAGE), and low-density lipoprotein receptor-related protein 1 (LRP1) in hCMEC/D3 cells were examined using western blotting and immunofluorescent staining. The mRNA levels of occludin, RAGE, and LRP1 were measured using quantitative real-time polymerase chain reaction. HIV-1 Tat at 1 µg/mL and the Rho inhibitor hydroxyfasudil (HF) at 30 µmol/L, with 24 h exposure, had no significant effect on hCMEC/D3 cell viability. Treatment with HIV-1 Tat protein decreased mRNA and protein levels of occludin and LRP1 and upregulated the expression of RAGE; however, these effects were attenuated by HF. These data suggest that the Rho/ROCK signaling pathway is involved in HIV-1 Tat-mediated changes in occludin, RAGE, and LRP1 in hCMEC/D3 cells. HF may have a beneficial influence by protecting the integrity of the blood-brain barrier and the expression of Aβ transfer receptors.
HIV-1反式激活蛋白（Tat）已被证明在与HIV相关的神经认知障碍中起重要作用。本研究的目的是在HIV-1相关病理学背景下，评估人脑血管内皮细胞（hCMEC/D3）中闭合蛋白与淀粉样β蛋白（Aβ）转运受体之间的关系。采用蛋白质印迹法和免疫荧光染色法检测hCMEC/D3细胞中闭合蛋白、晚期糖基化终末产物受体（RAGE）和低密度脂蛋白受体相关蛋白1（LRP1）的蛋白表达。采用定量实时聚合酶链反应测定闭合蛋白、RAGE和LRP1的mRNA水平。1μg/mL的HIV-1 Tat和30μmol/L的Rho抑制剂羟基法舒地尔（HF），作用24小时，对hCMEC/D3细胞活力无显著影响。用HIV-1 Tat蛋白处理可降低闭合蛋白和LRP1的mRNA和蛋白水平，并上调RAGE的表达；然而，这些作用被HF减弱。这些数据表明，Rho/ROCK信号通路参与了HIV-1 Tat介导的hCMEC/D3细胞中闭合蛋白、RAGE和LRP1的变化。HF可能通过保护血脑屏障的完整性和Aβ转运受体的表达而产生有益影响。
