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目的:探讨血管紧张素Ⅱ(AngⅡ)/乳脂肪球表皮生长因子8(MFG-E8)信号通路在人脐静脉内皮细胞(HUVECs)基质重构调控中的分子生物学机制。方法:利用实时定量聚合酶链反应(RT-qPCR)实验对AngⅡ和MFG-E8浓度进行筛选,根据实验需求在AngⅡ和MFG-E8浓度筛选实验中分为5组,阴性对照组(DPBS)、AngII 0.1 μmol/L、AngII 1.0 μmol/L、MFG-E8 50.0 μg/L、MFG-E8 100.0 μg/L。并利用RT-qPCR和蛋白质印迹法(Western blot)检测AngⅡ和MFG-E8的相互作用关系及对下游基因单核细胞趋化蛋白-1(MCP-1)、基质金属蛋白酶-2(MMP-2)和转化生长因子-β1(TGF-β1)表达的影响。两组间比较采用n t检验,多组间比较采用单因素方差分析,组间两两比较采用SNK-n q检验。n 结果:在脐静脉内皮细胞中,1 μmol/L的AngⅡ或100 μg/L MFG-E8能够促进AngⅡ mRNA的表达(1.62±0.30、2.25±0.36比1.00±0.00,n t=3.606、6.015,n P<0.05),差异有统计学意义。0.1 μmol/L或1.0 μmol/L的AngⅡ(1.27±0.11、1.58±0.14比1.00±0.00,n t=4.320、6.887,n P<0.05),以及50.0 μg/L或100.0 μg/L的MFG-E8能够促进MFG-E8的表达(1.90±0.09、2.77±0.07比1.00±0.00,n t=17.240、43.130,n P<0.05),差异有统计学意义。通过AngⅡ刺激HUVECs细胞建立血管高压模型,结果显示,AngⅡ能够显著促进HUVECs内AngⅡ、MFG-E8以及TGF-β1 mRNA的表达(1.62±0.13、4.27±0.47、1.56±0.11比1.00±0.00,n t=7.911、12.060、8.710,n P<0.01),而AngⅡ受体拮抗剂洛沙钽能抑制AngⅡ的作用,AngⅡ与MFG-E8共处理,则能进一步促进AngⅡ的作用(1.86±0.21比1.00±0.00,n t=7.023,n P<0.01),显著协同促进MCP-1和MMP-2 mRNA的表达(1.69±0.25、1.69±0.15比1.00±0.00,n t=4.735、7.845,n P<0.01),差异均有统计学意义。此外,AngⅡ能够显著促进MFG-E8蛋白水平的表达(0.23±0.01比0.16±0.03,n t=3.895,n P<0.05),而洛沙钽能抑制AngⅡ对MFG-E8蛋白表达的促进作用,AngⅡ与MFG-E8共处理,则能够进一步促进AngⅡ对MFG-E8(0.22±0.03比0.16±0.03,n t=2.493,n P<0.05)和TGF-β1蛋白水平表达的促进作用(0.66±0.06比0.50±0.03,n t=3.870,n P<0.01),差异均有统计学意义。n 结论:AngⅡ能够促进MFG-E8的表达,而MFG-E8也能够促进AngⅡ的表达,AngⅡ/MFG-E8信号通路能够促进基质重构标志物MCP-1、MMP-2和TGF-β1的表达。“,”Objective:To explore the effect of Angiotensin Ⅱ (AngⅡ)/milk fat globule epidermal growth factor 8 (MFG-E8) signaling on matrix remodeling of human umbilical vein endothelial cells (HUVECs) and underlying molecular mechanism.Methods:Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to screen the working concentration of AngⅡ and MFG-E8. The correlation of AngⅡ with MFG-E8 and the effect of AngⅡ/MFG-E8 signaling on monocyte chemotactic protein 1 (MCP-1), matrix metalloproteinase-2 (MMP-2), and transforming growth factor-β1 (TGF-β1) expression were analyzed by RT-qPCR and Western blotting. All data were analyzed by SPSS 17.0. Then t test was used for pairwise comparison between multiple groups.n Results:In HUVECs, 1 μmol/L AngⅡ or 100 μg/L MFG-E8 could significantly enhance AngⅡ mRNA expression (1.62±0.30, 2.25±0.36 vs. 1.00±0.00, n t=3.606, 6.015, n P<0.05). 0.1 μmol/L or 1.0 μmol/L AngⅡ (1.27±0.11, 1.58±0.14 vs. 1.00±0.00,n t=4.320, 6.887, n P<0.05), and 50.0 or 100.0 μg/L MFG-E8 could significantly increase the expression of MFG-E8 mRNA (1.90±0.09, 2.77±0.07 vs. 1.00±0.00,n t=17.240, 43.130, n P<0.05). AngⅡ stimulation was used to establish a vascular hypertension model in HUVECs, and the data showed that AngⅡ significantly up-regulated the mRNA expression of AngⅡ, MFG-E8 and TGF-β1 (1.62±0.13, 4.27±0.47, 1.56±0.11 vs. 1.00±0.00,n t=7.911, 12.060, 8.710, n P<0.01), while co-treatment of AngⅡ with AngⅡ receptor antagonist losartan potassium could inhibit this phenotype. Co-treatment of AngⅡ and MFG-E8 further promoted the expression of MCP-1 and MMP-2 mRNA (1.69±0.25, 1.69±0.15 vs. 1.00±0.00,n t=4.735, 7.845, n P<0.01). In addition, AngⅡ could significantly increase the expression of MFG-E8 in protein level (0.23±0.01 vs. 0.16±0.03,n t=3.895, n P<0.05), while co-treatment of AngⅡ and losartan potassium inhibited the effect of AngⅡ. Co-treatment of AngⅡ and MFG-E8 further promoted the effect of AngⅡ.n Conclusion:AngⅡ can enhance the expression of MFG-E8, and MFG-E8 can promote the expression of AngⅡ in HUVECs. The AngⅡ/MFG-E8 signaling increases the expression of matrix remodeling markers, including MCP-1, MMP-2 and TGF-β1. These findings provide a new insight into the mechanism by which AngⅡ/MFG-E8 signaling regulates arterial remodeling. The AngⅡ/MFG-E8 signaling pathway may provide new targets and strategies for the treatment of atherosclerosis.