摘要
目的探讨特异性抗体P210对氧化低密度脂蛋白(Ox-LDL)诱导的血管内皮细胞脂质蓄积和内皮连接屏障损伤的影响。方法通过Transwell小室建立人脐静脉内皮细胞(HUVEC)与人单核细胞(THP-1)来源巨噬细胞共孵育模型,利用激光共聚焦显微镜观察Ox-LDL及特异性抗体对内皮细胞及内膜下巨噬细胞脂质蓄积的影响;将HUVEC随机分为3组:对照组、Ox-LDL组、Ox-LDL+Ab(特异性抗体)组,分别用DMEM完全培养液,Ox-LDL(50μg/mL,100μg/mL),Ox-LDL(50μg/mL)+Ab(100 ng/mL,200 ng/mL)处理24 h。Western blot检测各组细胞植物血凝素样氧化低密度脂蛋白受体-1(LOX-1)、血管内皮钙粘蛋白(VE-Cadherin)、紧密连接蛋白(Occludin)、单核细胞趋化因子-1(MCP-1)、血管细胞粘附分子-1(VCAM-1)蛋白的表达。结果Transwell实验中Ox-LDL组小室上、下腔的细胞内均有较多荧光探针标记的Ox-LDL蓄积,在抗体组中这一效应被显著抑制。与对照组比较,Ox-LDL组的LOX-1、MCP-1和VCAM-1表达明显增高,VE-Cadherin、Occludin的表达明显降低;与Ox-LDL组比较,抗体组的LOX-1、MCP-1和VCAM-1的表达明显下降,VE-Cadherin、Occludin的表达明显升高。结论特异性抗体P210包被后可以明显减少内皮细胞和巨噬细胞内脂质蓄积,减轻Ox-LDL诱导的HUVEC损伤,保护内皮细胞连接屏障。
Objective To investigate the effect of specific antibody P210 on oxidized low-density lipoprotein(Ox-LDL)-induced vascular endothelial cell lipid accumulation and endothelial junction barrier injury.Methods A co-incubation model of HUVEC and THP-1 derived macrophages was established using a Transwell chamber.The effects of Ox-LDL and specific antibodies on lipid accumulation in endothelial cells and subintimal macrophages were observed by laser scanning confocal microscopy.HUVECs were randomly divided into three groups:control group,Ox-LDL group,Ox-LDL+Ab(specific antibody)group,cells in which were respectively treated with DMEM complete medium,Ox-LDL(50μg/mL,100μg/mL),Ox-LDL(50μg/mL)and antibody(100 ng/mL,200 ng/mL)for 24 h.The protein expression levels of LOX-1,VE-Cadherin,Occludin,MCP-1 and VCAM-1 were detected by Western blotting.Results In the Transwell experiment,cells in the upper and lower chambers of Ox-LDL group had more fluorescence-labeled Ox-LDL accumulations and this effect was significantly inhibited in antibody group.Compared with the control group,the protein expressions of LOX-1,MCP-1 and VCAM-1 in the Ox-LDL-induced group were significantly increased,and the protein expression levels of VE-Cadherin and Occludin were significantly decreased.Compared with the Ox-LDL-induced group,the protein expression levels of LOX-1,MCP-1 and VCAM-1 were significantly down-regulated while the protein expression levels of VE-Cadherin and Occludin were up-regulated in antibody group.Conclusion Specific antibody P210 can significantly reduce lipid accumulation in endothelial cells and macrophages,alleviate Ox-LDL induced vascular endothelial cell injury,and protect the endothelial cell junction barrier.
作者
陈娟
王志文
魏宇淼
Chen Juan;Wang Zhiwen;Wei Yumiao(Department of Cardiology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2021年第1期1-6,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81873491,No.82070376)。
关键词
P210抗体
氧化低密度脂蛋白
内皮细胞损伤
脂质蓄积
P210 antibody
oxidized low-density lipoprotein
endothelial cell injury
lipid accumulation
