【摘要】 目的探讨甲酰肽受体(formyl peptide receptor,FPR)在缺氧时对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)炎症因子产生的作用。方法按照缺氧时间将大鼠血管平滑肌细胞随机分为0 h、12 h、24 h组,将各组细胞放入缺氧培养箱(1%O2、5%CO2、99%N2)培养相应时间后,用Western blot检测平滑肌细胞中IL-6、IL-8的蛋白水平,用RT-PCR检测其m RNA水平。同时使用FPR拮抗剂t BOC预处理细胞并缺氧不同时间(0 h、12 h、24 h、)后,检测IL-6、IL-8的表达水平。另外,为了进一步明确FPR与炎症因子表达的关系,用FPR激动剂f MLP直接处理细胞之后,检测IL-6、IL-8的表达水平。然后使用细胞在缺氧条件下培养后的上清液作用于未缺氧的细胞,检测IL-6、IL-8的表达水平变化,并通过观察FPR拮抗剂处理后的变化,明确FPR在其中的作用。用f MLP刺激p38抑制剂预处理过的细胞,检测IL-6、IL-8的表达水平变化,以观察FPR激活后引起炎症因子表达对p38途径的依赖性。结果缺氧后血管平滑肌细胞的IL-6、IL-8 m RNA和蛋白含量均显著高于0 h组;当t BOC处理细胞后显著削弱了这一效应(P<0.05)。而且FPR激动剂能直接显著提高未缺氧的细胞的IL-6、IL-8的表达水平(P<0.05)。使用缺氧细胞的上清作用于常氧细胞后IL-6、IL-8的表达水平显著高于常氧细胞上清的作用效果;FPR拮抗剂处理显著削弱了这一效应(P<0.05)。用p38抑制剂预处理过的细胞加入f MLP与单纯加入f MLP的细胞相比,血管平滑肌细胞IL-6、IL-8表达水平显著低于单纯加入f MLP组(P<0.05)。结论缺氧引起血管平滑肌细胞IL-6、IL-8的表达,FPR在这一过程中发挥了重要的介导作用,而FPR是被缺氧细胞释放出来的激动剂激活。激活后的FPR引起IL-6、IL-8的表达依赖于p38信号途径。更多还原

【Abstract】 This study was performed to investigate the role of Formyl peptide receptor(FPR) in the production of inflammatory cytokines from vascular smooth muscle cells(VSMCs) during hypoxia. The m RNA levels of IL-6 and IL-8 in VSMCs of hypoxia rats were examined with RT-PCR, while the protein levels of IL-6 and IL-8 in VSMCs of hypoxia rats were detected by Western blotting. The m RNA and protein levels of IL-6 and IL-8 in vascular smooth muscle cells of hypoxia group were significantly higher than those in control group, and this effect was significantly attenuated by t BOC treatment(P<0.05). Moreover, FPR agonist directly and significantly upregulated the expression of IL-6 and IL-8 in non-hypoxic cells(P<0.05). The expression levels of IL-6 and IL-8 significantly higher than that in the cells stimulated by normoxic supernatant, while FPR antagonist treatment could significantly reduced the difference(P<0.05).The f MLP-induced IL-6 and IL-8 expression in VSMCs were significantly down-regulated by p38 inhibitor(P<0.05). In conclusion, hypoxia induces the expression of IL-6 and IL-8 in vascular smooth muscle cells. FPR plays an important mediating role in this process. FPRs can be activated by some agonists released by hypoxic cells. And the upregulation of IL-6 and IL-8 expression by activated FPRs is dependent on the p38 signaling pathway.更多还原