武汉科技大学附属天佑医院心血管内科;华中科技大学同济医学院附属武汉儿童医院(武汉市妇幼保健院)儿童保健科;华中科技大学同济医学院;华中科技大学同济医学院附属同济医院心血管内科;
目的 探究CD151通过调控囊泡内化再循环对血管通透性的影响及机制。方法 野生型小鼠和CD151敲除小鼠分别分为WT-con组、WT-模型组、KO-con组和KO-模型组，每组6只。WT-模型组和KO-模型组采用腹腔注射脂多糖(LPS)制备脓毒症急性肺损伤(ALI)模型，WT-con组和KO-con组腹腔注射磷酸盐缓冲液(PBS)作为对照。造模后24 h,通过Miles实验测定肺血管通透性；将沉默CD151表达的siRNA(si-CD151)和阴性对照si-NC转入EA.hy 926细胞，分别观察基础条件及血管内皮生长因子(VEGF-A)刺激条件下，内皮细胞层在不同时间点对FITC-dextran的通透性；CD151敲低组和对照组内皮细胞行转录组测序；免疫荧光检测各组细胞CD151分布及内化；Western bolt及RT-qPCR检测CD151敲低组及对照组VE-cadherin表达情况，免疫荧光检测各组细胞VE-cadherin分布及内化。结果 Miles实验结果提示，WT-模型组小鼠肺组织中染料渗出相较WT-con组增加(P<0.01),KO-模型组小鼠肺组织中染料渗出相较WT-模型组增加(P<0.05)。内皮细胞层通透性检测结果提示，在VEGF-A刺激30、60和120 min后，CD151敲低组对FITC-dextran的通透性显著高于对照组(P<0.05)。转录组测序结果提示，内皮细胞中CD151与囊泡介导的转运存在密切关联。Western bolt和RT-qPCR结果提示，CD151敲低的内皮细胞中VE-cadherin在蛋白和mRNA水平表达较对照组显著下降(均P<0.01)。免疫荧光染色结果提示，在VEGF-A刺激后，与对照组比较，CD151表达下降显著破坏细胞-细胞连接处VE-cadherin的表达，降低了CD151和VE-cadherin在核周区域的共定位。结论 CD151的缺失影响内皮细胞囊泡内化再循环，影响VE-cadherin的表达与内化活动，进而影响血管通透性。
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[1] Long M E,Mallampalli R K,Horowitz J C.Pathogenesis of pneumonia and acute lung injury[J].Clin Sci (Lond),2022 ,136(10):747-69.doi:10.1042/CS20210879.
[2] Jiang J,Huang K,Xu S,et al.Targeting NOX4 alleviates sepsis-induced acute lung injury via attenuation of redox-sensitive activation of CaMKII/ERK1/2/MLCK and endothelial cell barrier dysfunction[J].Redox Biol ,2020,36:101638 .doi:10.1016/j.redox.2020.101638.
[3] Zhang F,Michaelson J E,Moshiach S,et al.Tetraspanin CD151 maintains vascular stability by balancing the forces of cell adhesion and cytoskeletal tension[J].Blood ,2011,118(15):4274-84.doi:10.1182/blood-2011-03-339531.
[4] Mukherjee S,Tessema M,Wandinger-Ness A.Vesicular trafficking of tyrosine kinase receptors and associated proteins in the regulation of signaling and vascular function[J].Circ Res ,2006,98(6):743-56.doi:10.1161/01.RES.0000214545.99387.e3.
[5] Kuna R S,Field S J.GOLPH3:a Golgi phosphatidylinositol(4)phosphate effector that directs vesicle trafficking and drives cancer[J].J Lipid Res ,2019,60(2):269-75.doi:10.1194/jlr.R088328.
[6] Clark P R,Kim R K,Pober J S,et al.Tumor necrosis factor disrupts claudin-5 endothelial tight junction barriers in two distinct NF-κB-dependent phases[J].PLoS One,2015,10(3):e0120075.doi:10.1371/journal.pone.0120075.
[7] Lindstedt S,Wang Q,Niroomand A,et al.High resolution fluorescence imaging of the alveolar scaffold as a novel tool to assess lung injury[J].Sci Rep ,2024,14(1):6662.doi:10.1038/s41598-024-57313-6.
[8] Inagawa R,Okada H,Takemura G,et al.Ultrastructural alteration of pulmonary capillary endothelial glycocalyx during endotoxemia[J].Chest ,2018,154(2):317-25.doi:10.1016/j.chest.2018.03.003.
[9] Xu W,Gao C,Wu J.CD151 alleviates early blood-brain barrier dysfunction after experimental focal brain ischemia in rats[J].Cell Mol Neurobiol ,2021,41(1):151-62.doi:10.1007/s10571-020-00842-1.
[10] De Franceschi N,Hamidi H,Alanko J,et al.Integrin traffic - the update[J].J Cell Sci ,2015,128(5):839-52.doi:10.1242/jcs.161653.
[11] Cho H D,Nhàn N T T,Zhou C,et al.KIF13B mediates VEGFR2 recycling to modulate vascular permeability[J].Cell Mol Life Sci ,2023,80(4):91.doi:10.1007/s00018-023-04752-5.
[12] Qiao X,Yin J,Zheng Z,et al.Endothelial cell dynamics in sepsis-induced acute lung injury and acute respiratory distress syndrome:pathogenesis and therapeutic implications[J].Cell Commun Signal ,2024,22(1):241.doi:10.1186/s12964-024-01620-y.
[13] Méndez-Barbero N,San Sebastian-Jaraba I,Blázquez-Serra R,et al.Annexins and cardiovascular diseases:beyond membrane trafficking and repair[J].Front Cell Dev Biol ,2022,10:1000760.doi:10.3389/fcell.2022.1000760.
[14] Rana S,Claas C,Kretz C C,et al.Activation-induced internalization differs for the tetraspanins CD9 and TSPAN8:impact on tumor cell motility[J].Int J Biochem Cell Biol ,2011,43(1):106-19.doi:10.1016/j.biocel.2010.10.002.
[15] Wu F T,Paez-Ribes M,Xu P,et al.Aflibercept and Ang1 supplementation improve neoadjuvant or adjuvant chemotherapy in a preclinical model of resectable breast cancer[J].Sci Rep,2016,6:36694.doi:10.1038/srep36694.
基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2025.02.005
中图分类号:R563
引用信息:
[1]范诗浪,蒋卢映,章子璇等.CD151通过囊泡内化再循环调控血管通透性的机制[J].安徽医科大学学报,2025,60(02):218-225+233.DOI:10.19405/j.cnki.issn1000-1492.2025.02.005.
基金信息:
国家自然科学基金(编号：81873535); 湖北省自然科学基金(编号：2020CFB573)~~