Aliskiren抑制LPS诱导HUVECs新生血管的形成及可能机制

 目的: 研究阿利吉仑(aliskiren)对脂多糖(LPS)诱导人脐静脉内皮细胞(HUVECs)新生血管形成能力的影响及可能的机制。方法: 常规培养的HUVECs随机分为空白组和肾素组,ELISA法测定炎性细胞因子肿瘤坏死因子-α(TNF-α)和细胞间黏附分子-1(ICAM-1)水平,Western blot法检测Toll样受体4(TLR4)和ICAM-1的蛋白水平。将常规培养的HUVECs随机分为空白对照组、LPS模型组以及aliskiren低剂量(1μ mol/L)、中剂量(10μ mol/L)和高剂量(100μ mol/L)组。MTT法和BrdU法检测HUVECs的增殖能力,Transwell法测定HUVECs的迁移率,以HUVECs在Matrigel胶上形成管腔结构情况来判断其血管形成能力。ELISA测定炎性细胞因子TNF-α、ICAM-1和单核细胞趋化蛋白-1(MCP-1)的水平,RT-PCR和Western blot法检测肾素、TLR4、基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)的mRNA和蛋白水平。结果: 肾素能够刺激HUVECs炎症因子的分泌及TLR4的表达;aliskiren呈浓度依赖性抑制HUVECs增殖、迁移及新生血管形成,降低MCP-1、TNF-α、IL-6水平及肾素、MMP-2、MMP-9的表达,抑制TLR4表达(P<0.05)。结论: Aliskiren能够有效抑制LPS诱导HUVECs新生血管形成能力,可能与其下调肾素表达抑制TLR4途径介导的炎症反应及MMP-2、MMP-9生成有关。

Inhibitory effect of aliskiren on LPS-induced angiogenesis of HUVECs

AIM: To investigate the inhibitory effect of aliskiren on the angiogenesis of human umbilical vein endothelial cells(HUVECs) induced by lipopolysaccharide(LPS) and to explore its possible mechanism. METHODS: HUVECs were cultured and randomly divided into blank group and renin group. The levels of tumor necrosis factor-α(TNF-α) and intercellular adhesion molecule-1(ICAM-1) in the culture supernatant were detected by ELISA. The protein levels of Toll-like receptor 4(TLR4) and ICAM-1 in the HUVECs were determined by Western blot. HUVECs were cultured and randomly divided into control group, LPS group, low-dose(1μmol/L) aliskiren group, middle-dose(10μmol/L) aliskiren group and high-dose(100μmol/L) aliskiren group. The proliferation of HUVECs was detected by MTT and BrdU assays. The mobility of HUVECs was measured by Transwell assay. The formation of the vessels was judged by observing the formation of the luminal structure by HUVECs in Matrigel. The levels of TNF-α, ICAM-1 and monocyte chemotactic protein 1(MCP-1) in the culture supernatant were measured by ELISA. The expression of renin, TLR4, matrix me-talloproteinases-2(MMP-2) and matrix metalloproteinases-9(MMP-9) at mRNA and protein levels in the HUVECs was determined by RT-PCR and Western blot. RESULTS: Renin stimulated the expression of inflammatory factors and TLR4 in the HUVECs. Aliskiren inhibited the growth, migration and angiogenesis of HUVECs in a dose-dependent manner, decreased the levels of TNF-α, ICAM-1 and MCP-1 and the expression of renin, MMP-2 and MMP-9, and inhibited TLR4 expression(P<0.05).CONCLUSION: Aliskiren inhibits LPS-induced angiogenesis of HUVECs, which may be related to the down-regulation of renin expression, the inhibition of TLR4-mediated inflammatory reaction, and the formation of MMP-9 and MMP-2.