隐丹参酮对TNF-α诱导的人脐静脉内皮细胞粘附分子ICAM-1表达的作用

The effect of cryptotanshinone on the expression of ICAM-1 in TNF-α-induced human umbilical vein endothelial cells

目的:探讨隐丹参酮对TNF-α诱导的人脐静脉内皮细胞(HUVEC)粘附分子ICAM-1表达的影响及机制。方法:通过建立TNF-α诱导的HUVEC细胞炎症模型,以抗氧化剂吡咯烷二硫代氨基甲酸盐(PDTC)做为阳性对照,用细胞酶联免疫吸附法、免疫荧光法测定HUVEC表面ICAM-1的表达,用实时定量PCR(RT-PCR)法检测ICAM-1的mRNA表达,用免疫荧光法检测核转录因子NF-κBp65蛋白的核转运。结果:隐丹参酮(3μM,1μM)可明显降低TNF-α诱导的HUVEC细胞表面粘附分子ICAM-1增多;明显抑制ICAM-1的基因表达;并对TNF-α诱导的HUVEC细胞核转录因子NF-κBp65蛋白的核转移有抑制作用。结论:隐丹参酮可剂量依赖性的抑制TNF-α诱导的HUVEC细胞粘附分子ICAM-1的基因表达以发挥抗动脉粥样硬化的作用,该作用的机制可能与其抑制转录因子NF-κBp65蛋白的核转运,从而抑制了NF-κB的激活有关。

Objective： To observe the effect of cryptotanshinone on the expression of ICAM-1 in the TNF-alpha-induced human umbilical vein endothelial cells （HUVECs）. Methods： First, the cytotoxicity of cryptotanshinone on HUVECs were evaluated by XTT cell proliferation assay kit. Then, TNF-α induced HUVECs inflammatiom model was established to analyze the expression of ICAM-1 by the method of cell-ELISA and immunofluorescent staining , and the mRNA expression of ICAM-1 by the method of RT-PCR. Moreover,the nuclear translocation of NF-kB p65 protein was studied also by the method of immunofluorescent staining. Results： Cryptotanshinone can significantly inhibit the upregulation of the expression of ICAM-1 on the surface of HUVECs and the mRNA expression of ICAM-1 in the cells induced by TNF-α. We also find the nuclear translocation of NF-kB p65 protein was inhibited by the treatment of cryptotanshinone dose dependently, especially at the concentration of 3μM and 1 μM. Conclusions： These results suggest that eryptotanshinone has an anti-inflammatory effect by inhibiting the expression of ICAM-1 and the nuclear translocation of NF-kB p65 protein, and may play an important role in the prevention of atherosclerosis and inflammation in vivo.