Molecular Biology Unit, Faculty of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, India.
Molecular Biology Unit, Faculty of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, India.
Cell Signal. 2023 Jun;106:110659. doi: 10.1016/j.cellsig.2023.110659. Epub 2023 Mar 21.
Zika virus infection has been reported to cause microcephaly in newborns. ZIKV exploits various strategies to cross the blood-brain barrier. ZIKV NS1 may compromise the barrier integrity of endothelial cells by regulating expression of junctional proteins. MicroRNAs play an important role in post-transcriptional gene regulations. We demonstrated that ZIKV-NS1 affected the adherence junction protein in human brain microvascular endothelial cells via hsa-miR-29b-3p/DNMT3b/MMP-9 pathway. The hCMEC/D3 cells were exposed to ZIKV-NS1 with different doses (500 ng/mL and 1000 ng/mL) for 24 h. The expression pattern of DNTM3b, MMP-9, and VE-cadherin were studied using immunoblotting and the distribution of DNMT3b and MMP-9 were studied using immunofluorescence. The quantification of hsa-miR-29b-3p was done through qRT-PCR. Direct regulation of DNMT3b by hsa-miR-29b-3p was demonstrated by overexpression of hsa-miR-29b-3p using hsa-miR-29b-3p mimic, and knockdown of hsa-miR-29b-3p by using hsa-miR-29b-3p inhibitors. The ZIKV-NS1 affected the barrier function of endothelial cells through the increased expression of hsa-miR29b-3p, which suppressed the DNMT3b, thus enhanced expression of MMP-9, which finally suppressed the expression of VE-cadherin. These findings suggested that ZIKV-NS1 alters the expression of Adherens Junction protein in human brain microvascular endothelial cells through hsa-miR-29b-3p/DNMT3b/MMP-9 pathway, which compromised the barrier function of human brain microvascular endothelial cells.
寨卡病毒感染已被报道可导致新生儿小头畸形。寨卡病毒利用各种策略穿过血脑屏障。寨卡病毒 NS1 可能通过调节连接蛋白的表达来损害内皮细胞的屏障完整性。微小 RNA 在转录后基因调控中发挥重要作用。我们证明，寨卡病毒 NS1 通过 hsa-miR-29b-3p/DNMT3b/MMP-9 通路影响人脑微血管内皮细胞中的粘着连接蛋白。用不同剂量（500ng/mL 和 1000ng/mL）的 ZIKV-NS1 处理 hCMEC/D3 细胞 24 小时。使用免疫印迹研究 DNTM3b、MMP-9 和 VE-cadherin 的表达模式，并使用免疫荧光研究 DNMT3b 和 MMP-9 的分布。通过 qRT-PCR 定量 hsa-miR-29b-3p。通过使用 hsa-miR-29b-3p 模拟物过表达 hsa-miR-29b-3p，以及使用 hsa-miR-29b-3p 抑制剂敲低 hsa-miR-29b-3p，证明 hsa-miR-29b-3p 对 DNMT3b 的直接调节作用。ZIKV-NS1 通过增加 hsa-miR29b-3p 的表达影响内皮细胞的屏障功能，抑制 DNMT3b，从而增强 MMP-9 的表达，最终抑制 VE-cadherin 的表达。这些发现表明，ZIKV-NS1 通过 hsa-miR-29b-3p/DNMT3b/MMP-9 通路改变人脑微血管内皮细胞中粘着连接蛋白的表达，从而损害人脑微血管内皮细胞的屏障功能。
