Zika virus (ZIKV) is a neurotropic virus that causes microcephaly in newborns and Guillain-Barré syndrome (GBS) in adults. ZIKV is known to transmigrate through the blood–brain barrier (BBB) by utilizing different strategies. NS1 is a conserved flavivirus protein, which is secreted extracellularly. ZIKV-NS1 has been shown to target adherens junctions (AJs) and tight junctions (TJs) to disrupt the endothelial barrier integrity. The microRNAs are short non-coding RNAs, which post-transcriptionally regulate the gene expression by binding to 3’ UTR of the target gene. In the present study, we studied the ZIKV-NS1-mediated effect through hsa-miR-101-3p on the junctional barrier integrity in human brain microvascular endothelial cells. We exposed hBMVECs and hCMEC/D3 cells with ZIKV-NS1 at different time points (12 h and 24 h) with the doses 500 ng/mL and 1000 ng/mL. The change in the expression of VE-cadherin and claudin-5 was quantified using immunoblotting. The expression of the hsa-miR-101-3p was quantified using qRT-PCR. To prove the targeting of hsa-miR-101-3p to VE-cadherin, we transfected hsa-miR-101-3p mimic, scramble, hsa-miR-101-3p inhibitor, and Cy3 in the ZIKV-NS1-exposed hCMEC/D3 cells. The distribution and expression of the VE-cadherin and claudin-5 were observed using immunofluorescence and immunoblotting. The ZIKV-NS1 compromises the endothelial barrier integrity by disrupting the VE-cadherin and claudin-5 protein expression via hsa-miR-101-3p. The findings of this study suggest that ZIKV-NS1 dysregulates the adherens junction and tight junction proteins through hsa-miR-101-3p, which compromises the barrier integrity of human brain microvascular endothelial cells.

寨卡病毒 (ZIKV) 是一种嗜神经病毒，可导致新生儿小头畸形和成人格林巴利综合征 (GBS)。众所周知，ZIKV 通过利用不同的策略穿越血脑屏障 (BBB)。NS1 是一种保守的黄病毒蛋白，在细胞外分泌。ZIKV-NS1 已被证明可以靶向粘附连接 (AJ) 和紧密连接 (TJ)，以破坏内皮屏障的完整性。microRNA 是短的非编码 RNA，通过与靶基因的 3' UTR 结合，在转录后调节基因表达。在本研究中，我们通过 hsa-miR-101-3p 研究了 ZIKV-NS1 介导的对人脑微血管内皮细胞连接屏障完整性的影响。我们在不同时间点（12 小时和 24 小时）以 500 ng/mL 和 1000 ng/mL 的剂量用 ZIKV-NS1 暴露 hBMVEC 和 hCMEC/D3 细胞。使用免疫印迹定量 VE-cadherin 和 claudin-5 表达的变化。使用 qRT-PCR 量化 hsa-miR-101-3p 的表达。为了证明 hsa-miR-101-3p 对 VE-cadherin 的靶向性，我们在暴露于 ZIKV-NS1 的 hCMEC/ D3 细胞。使用免疫荧光和免疫印迹观察VE-cadherin和claudin-5的分布和表达。ZIKV-NS1 通过 hsa-miR-101-3p 破坏 VE-cadherin 和 Claudin-5 蛋白的表达，从而损害内皮屏障的完整性。