oxLDL/β2GPⅠ/aβ2GPⅠ复合物通过TLR4/MyD88/NF-κB途径促进血管内皮细胞血管生成  

Mechanism of oxLDL/β2GPⅠ/aβ2GPⅠcomplex promoting the angiogenesis in vascular endothelial cells through TLR4//MyD88/NF-κB signaling

作　　者：张贵婷 何超[2] ZHANG Guiting;HE Chao(Department of Laboratory Medicine,Nanjing Drum Tower Hospital Clinical College of Nanjing University,Nanjing 210008,China;Center Laboratory,the Fourth Affiliated Hospital of Jiangsu University)

机构地区：[1]南京大学医学院附属鼓楼医院检验科,210008 [2]江苏大学第四附属医院中心实验室

出　　处：《天津医药》2024年第11期1131-1136,共6页Tianjin Medical Journal

基　　金：江苏省卫生健康委科研项目(H2023033);镇江市重点研发计划社会发展项目(SH2022082)。

摘　　要：目的探讨氧化型低密度脂蛋白(oxLDL)/β2糖蛋白Ⅰ(β2GPⅠ)/抗β2糖蛋白Ⅰ抗体(aβ2GPⅠ)复合物对血管内皮细胞增殖、迁移及血管生成的影响及其机制。方法将人脐静脉内皮细胞(HUVEC)培养至对数生长期,分为对照组(正常培养)、oxLDL组(50 mg/L oxLDL)、oxLDL/β2GPⅠ/aβ2GPⅠ组(50 mg/L oxLDL/100 mg/Lβ2GPⅠ/100 mg/L aβ2GPⅠ)和血管内皮生长因子(VEGF)组(100μg/L VEGF)。实时荧光定量PCR(qPCR)法检测血管生成相关因子VEGF、血管内皮钙黏蛋白(VE-cadherin)、基质金属蛋白酶(MMP)-2及MMP-9的基因表达;CCK-8检测细胞增殖;划痕愈合实验和Transwell实验检测细胞的迁移和侵袭能力;基质胶管腔形成实验检测HUVEC的血管生成能力;Western blot法检测Toll样受体4(TLR4)、髓样分化因子88(MyD88)和核因子κB(NF-κB)的蛋白表达。结果与对照组相比,oxLDL/β2GPⅠ/aβ2GPⅠ组细胞增殖活力在处理48 h时增加;此外,与对照组相比,oxLDL/β2GPⅠ/aβ2GPⅠ组细胞迁移及血管生成能力增强,VEGF、VE-cadherin、MMP-2及MMP-9的mRNA水平升高(P<0.05),TLR4和MyD88蛋白水平及p-NF-κB p65/NF-κB p65比值升高(P<0.05)。结论oxLDL/β2GPⅠ/aβ2GPⅠ复合物可能通过TLR4/MyD88/NF-κB通路诱导血管内皮细胞增殖、迁移及血管生成。Objective To investigate effects of oxidized low density lipoprotein/β2 glycoprotein-Ⅰ/anti-β2 glycoprotein-Ⅰantibody(oxLDL/β2GPⅠ/aβ2GPⅠ)complex on the proliferation,migration and angiogenesis of vascular endothelial cells and its mechanism.Methods Human umbilical vein endothelial cells(HUVEC)were cultured to logarithmic growth phase and grouped into the control group(normal culture),the oxLDL group(50 mg/L oxLDL),the oxLDL/β2GPⅠ/aβ2GPⅠgroup(50 mg/L oxLDL/100 mg/Lβ2GPⅠ/100 mg/L aβ2GPⅠ)and the VEGF group(100μg/L VEGF).The gene expressions of VEGF,vascular endothelial cadherin(VE-cadherin),matrix metalloproteinase(MMP)-2 and MMP-9 were detected by real-time quantitative fluorescent PCR(qPCR).Cell counting kit-8(CCK-8)method was employed to detect cell proliferation.Cell migration and invasion were determined by scratch healing test and Transwell assay.Matrigel tube formation assay was used to observe the angiogenesis of HUVEC.The relative protein expression of TLR4,MyD88 and NF-κB were examined by Western blot assay.Results Compared with the control group,the proliferation activity of cells at 48 h of treatment was increased in the oxLDL/β2GPⅠ/aβ2GPⅠgroup(P<0.05).Moreover,compared with the control group,cell migration and angiogenesis were increased in the oxLDL/β2GPⅠ/aβ2GPⅠgroup,and the mRNA levels of VEGF,VE-cadherin,MMP-2 and MMP-9 were elevated(P<0.05).Compared with the control group,levels of TLR4 and MyD88 were elevated in the oxLDL/β2GPⅠ/aβ2GPⅠcomplex group(P<0.05),as well as levels of p-NF-κB p65/NF-κB p65(P<0.05).Conclusion oxLDL/β2GPⅠ/aβ2GPⅠcomplex may promote the proliferation,migration and tube formation of vascular endothelial cells by regulating TLR4/MyD88/NF-κB signaling pathway.

关 键 词：动脉粥样硬化 β2糖蛋白Ⅰ 新生血管化 病理性 细胞增殖 细胞运动 NF-ΚB 内皮细胞 氧化型低密度脂蛋白 

分 类 号：R392.9[医药卫生—免疫学]