Regulation on Toll-like Receptor 4 and Cell Barrier Function by Rab26 siRNA-loaded DNA Nanovector in Pulmonary Microvascular Endothelial Cells.
Regulation on Toll-like Receptor 4 and Cell Barrier Function by Rab26 siRNA-loaded DNA Nanovector in Pulmonary Microvascular Endothelial Cells.
复制标题
负载 Rab26 siRNA 的 DNA 纳米载体对肺微血管内皮细胞中 Toll 样受体 4 和细胞屏障功能的调节
作者：
Li H;He B;Liu X;Li J;Liu Q;Dong W;Xu Z;Qian G;Zuo H;Hu C;Qian H;Mao C;Wang G
小的GTP酶Rab26参与多种过程，如囊泡介导的分泌和自噬。然而，Rab26在人肺微血管内皮细胞(HPMVECs)中的作用机制尚不清楚。在本研究中，我们利用自组装的携带DNA Y基序的Rab26 siRNA纳米颗粒(siRab26-DYM)和Rab26腺病毒，深入研究了Rab26在HPMVECs通透性和凋亡中的作用及其新的机制。我们发现siRab26-DYM能够以时间和剂量依赖的方式有效地导入HPMVEC。重要的是，siRab26-DYM纳米载体通过促进Foxo1的核转位，进而激活Toll样受体4(Toll-like Receptor 4，TLR4)信号通路，显著加重了内毒素诱导的HPMVEC的凋亡和高通透性。Rab26腺病毒过表达可部分抑制脂多糖诱导的TLR4信号通路，抑制细胞凋亡，减轻HPMVECs的高通透性。以上结果提示，可通过调控Rab26来源的TLR4信号通路来调节HPMVECs的通透性和细胞凋亡，Rab26可作为治疗与内皮屏障功能相关的血管疾病的潜在靶点。
The small GTPase Rab26 is involved in multiple processes, such as vesicle-mediated secretion and autophagy. However, the mechanisms and functions of Rab26 in the human pulmonary microvascular endothelial cells (HPMVECs) are not clear. In this study, we thoroughly investigated the role and novel mechanism of Rab26 in permeability and apoptosis of HPMVECs using a self-assembled Rab26 siRNA loaded DNA Y-motif nanoparticle (siRab26-DYM) and Rab26 adenovirus. We found that siRab26-DYM could be efficiently transfected into HPMVECs in a time- and dose-dependent manner. Importantly, the siRab26-DYM nanovector markedly aggravated the LPS-induced apoptosis and hyper-permeability of HPMVECs by promoting the nuclear translocation of Foxo1, and subsequent activation of Toll-like receptor 4 (TLR4) signal pathway. Overexpression of Rab26 by Rab26 adenoviruses partially inactivated LPS-induced TLR4 signaling pathway, suppressed the cell apoptosis and attenuated the hyperpermeability of HPMVECs. These results suggest that the permeability and apoptosis of HPMVECs can be modulated by manipulating Rab26 derived TLR4 signaling pathway, and that Rab26 can be potential therapeutic target for the treatment of vascular diseases related to endothelial barrier functions.