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目的观察白藜芦醇对TNF-α刺激引起的内皮细胞炎症反应的影响,并围绕TNFR1探索相关作用机制。方法不同浓度(0.01、0.1、1、2.5、5、10、20、50、100、200、400 ng/ml)TNF-α处理人血管内皮细胞株EA.hy926,24 h后MTT法和ELISA法分别检测细胞增殖活力和细胞培养液中sICAM-1释放水平;白藜芦醇(浓度为0.1、1、10μmol/L)预处理24 h后,再用TNF-α(10 ng/ml)处理24 h,利用ELISA法检测细胞培养液中sICAM-1释放水平,qRT-PCR法检测ICAM-1mRNA和TNFR1 mRNA表达水平。结果高浓度TNF-α(≥100 ng/ml)刺激EA.hy926细胞后,细胞增殖活力显著下降(P<0.05),当TNF-浓度为200 ng/ml时,与对照组比较细胞增殖活力下降约59.7%;浓度为10 ng/ml的TNF-α刺激内皮细胞后,细胞的ICAM-1及TNFR1 mRNA表达明显上调(P<0.05);白藜芦醇预处理内皮细胞后可明显下调TNFR1 mRNA表达水平(P<0.05),并显著抑制TNF-α诱导的ICAM-1的表达(P<0.05),且抑制作用随着浓度的增加而增强。结论白藜芦醇可能通过抑制TNFR1表达,进而抑制TNF-α刺激引起的炎性因子释放,减轻内皮炎性损伤。
Objective To observe the effect of resveratrol on the inflammatory reaction of endothelial cells induced by TNF-α, and to explore the related mechanism of TNFR1. Methods The human vascular endothelial cell line EA.hy926 was treated with TNF-α at different concentrations (0.01,0.1,1,2.5,5,10,20,50,100,200,400 ng / ml) for 24 h, and MTT and ELISA The cell proliferation activity and sICAM-1 release in cell culture medium were detected respectively. Pretreatment with resveratrol at concentrations of 0.1, 1 and 10 μmol / L for 24 h followed by treatment with TNF-α (10 ng / ml) for 24 h h. The release of sICAM-1 in cell culture medium was detected by ELISA. The expression of ICAM-1 mRNA and TNFR1 mRNA was detected by qRT-PCR. Results The proliferation activity of EA.hy926 cells was significantly decreased (P <0.05) when high concentrations of TNF-α (≥100 ng / ml) were stimulated. When the concentration of TNF-α was 200 ng / ml, the cell proliferation activity decreased (59.7%). The expression of ICAM-1 and TNFR1 mRNA was up-regulated after TNF-α was stimulated with 10 ng / ml of TNF-α (P <0.05). Resveratrol pretreated endothelial cells could down-regulate the expression of TNFR1 mRNA (P <0.05), and significantly inhibited the expression of ICAM-1 induced by TNF-α (P <0.05), and the inhibitory effect increased with the increase of concentration. Conclusion Resveratrol may inhibit the release of inflammatory cytokines induced by TNF-α and decrease the inflammatory injury by inhibiting the expression of TNFR1.