目的：探讨辣椒素对脂多糖（ LPS）刺激后小鼠主动脉内皮细胞活化的影响，并探讨其可能机制。方法：（1）体外分离培养小鼠主动脉内皮细胞，免疫荧光鉴定内皮细胞特异性标志。（2）以100μg/L LPS作用于血管内皮细胞后，以不同浓度的辣椒素（50μmol/L、100μmol/L和200μmol/L）进行干预，分别于12、24和48 h收集主动脉血管内皮细胞及细胞上清液。采用ELISA法检测各组细胞上清液中的可溶性细胞间黏附分子1（ sICAM-1）、可溶性血管细胞黏附分子1（sVCAM-1）和可溶性P-选择素（sP-selectin）水平，Western blotting法检测各组主动脉血管内皮细胞核内NF-κB p65水平和胞浆p-IκBα、IκBα水平。结果：与对照组相比，LPS组细胞上清液中sP-se-lectin、sICAM-1和sVCAM-1含量显著升高（P＜0．05）， LPS能够时间依赖性上调sICAM-1和sVCAM-1的含量。与同时点的LPS组相比，辣椒素能够剂量依赖性下调sP-selectin、sICAM-1和sVCAM-1水平。与对照组相比，LPS作用24 h后，细胞核内NF-κB p65和胞浆p-IκBα蛋白水平显著升高（P＜0．05），胞浆IκBα蛋白水平显著降低（P＜0．05）。与同时点LPS组相比，辣椒素能够剂量依赖性下调细胞核内NF-κB p65和胞浆p-IκBα蛋白的水平（ P＜0.05），剂量依赖性上调胞浆IκBα蛋白水平（P＜0．05）。结论：辣椒素能够显著抑制LPS作用后血管内皮细胞活化水平，该效应可能是通过下调IκBα降解和NF-κB p65胞核内转位而实现的。%AIM:To investigate the effect of capsaicin on lipopolysaccharide ( LPS)-induced activation of cul-tured endothelial cells of mouse aorta in vitro.METHODS:The endothelial cells were isolated from mouse aorta and cul-tured in vitro, and the specific cell markers of the cells were identified by immunofluorescence staining.The cells were stimulated with LPS (100μg/L) combined with or without capsaicin, and the cells and supernatant were collected at 12 h, 24 h and 48 h.The levels of soluble intercellular adhesion molecule 1 ( sICAM-1) , soluble vascular cell adhesion molecule 1 (sVCAM-1) and soluble P-selectin (sP-selectin) in the supernatant were measured by ELISA.The levels of nuclear NF-κB p65 and cytopasmic p-IκBαand IκBαwere detected by Western blotting.RESULTS: Compared with control group, the levels of sP-selectin, sICAM-1 and sVCAM-1 in LPS group were significantly increased (P<0.05), and LPS promoted the expression of sICAM-1 and sVCAM-1 in a time-dependent manner.Compared with LPS group at the same time point, capsaicin inhibited the expression of sP-selectin, sICAM-1 and sVCAM-1 in a dose-dependent manner.Compared with con-trol group, the protein levels of NF-κB p65 and p-IκBαin LPS group at 24 h were significantly increased (P<0.05), while the protein level of IκBαin LPS group at 24 h were significantly decreased (P<0.05).Compared with LPS group, capsaicin decreased the protein levels of NF-κB p65 and p-IκBαand increased the protein level of IκBαin a dose-depend-ent manner.CONCLUSION:Capsaicin has a protective effect on LPS-induced vascular endothelial cell activation, which potentially contributes to the suppression of IκBαdegradation and NF-κB p65 nuclear translocation.
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