肌酸抑制TNF-α诱导内皮细胞黏附分子mRNA的表达

Effects of creatine on the expression of adhesion molecules and their mRNAs in cultured endothelial cells

目的探讨肌酸(creatine,CR)对血管内皮细胞(endothelail cells,EC)黏附分子E-selectin和ICAM-1及其mRNA表达的影响。方法以不同浓度的CR加至培养的血管EC24h,用TNF-α诱导EC活化3h,添加或未添加腺苷A2A受体拮抗剂(ZM241385),应用ELISA测定EC的E-selectin和ICAM-1的表达水平、用RT-PCR半定量E-selectin和ICAM-1mRNA的表达。结果EC经TNF-α诱导后,黏附分子及其mRNA表达明显增加,与对照组相比,E-selectin和ICAM-1分别增加82%和32%,E-selectin和ICAM-1的mRNA分别增加53%和24%。在添加CR0.5mmol/L组,这些表达明显下调,当CR添加浓度为5mmol/L时它们的表达接近对照组水平。CR对激活的CE黏附分子表达抑制作用能因ZM241385的添加而解除(P<0.05)。结论CR抑制活化了的ECE-Selectin和ICAM-1mRNR的表达,从而下调EC的E-selectin和ICAM-1的表达以发挥抗炎作用。

Objective Previous studies have shown that creatine （CR） can prevent WBC from adhering to blood vessel endothelium and decrease endothelial permeability induced by inflammatory mediators. To reveal potential mechanisms for the anti - inflammatory activity of CR, we investigated the effects of CR on the expression of adhesion molecules E -seleetin and/CAM - 1 and their mRNAs in cultured ECs. Methods Cultured endothelial cells （ECs） were supplemented with serial concentration of CR for 24 h. They were treated with TNF - α for 3 h, in the presence or absence of an A,2a receptor antagonist （ZM241385）. Concentreation of adhesion molecules E - selectin and ICAM - 1 in culture medium was measured with ELISA and their mRNA expression in the cells was detected with RT - PCR. Results TNF - α activation resulted in increased expression of adhesion molecules and their mRNAs. Compared to control group, TNF - α - activated ECs showed an 82% increase in E - selectin and 32% in ICAM - 1. E - selectin mRNA expression was increased by 24% and ICAM - 1 mRNA expression by 53%. These increased expressions could be brought down by supplementation of 0.5 mmoL/L CR to the TNF - α - activated ECs. Supplying 5 mmoL/L CR completely brought down the expression to control level. The CR inhibition effects on TNF - α induced expression of adhesion molecules and their mRNAs diminished when ZM241385 was added. Conclusion The present study suggests that CR may inhibit the expression of E - selectin and ICAM - 1 mRNAs and thus reduce the production of the adhesion molecules, As a result, inflammation may .be restrained.