葛根素对支气管上皮细胞和中性粒细胞共培养体系VCAM-1表达的影响

Effect of Puerarin on the Expression of VCAM-1 in Co-culture of Bronchial Epithelial Cells and Neutrophils in vitro

目的探讨葛根素抑制支气管上皮细胞(BEAS-2B)与中性粒细胞(NEU)相互作用调控气道炎症反应的机制。方法应用流式细胞仪(FCM)检测BEAS-2B细胞和NEU细胞表面血管细胞间黏附分子-1(VCAM-1)的蛋白质合成;应用实时PCR方法检测上述细胞中VCAM-1的基因表达。结果 BEAS-2B细胞和NEU细胞共同培养组中BEAS-2B表面的VCAM-1平均荧光强度显著高于BEAS-2B单独培养组,共同培养组中NEU表面的VCAM-1平均荧光强度也显著高于NEU单独培养组。共同培养组中BEAS-2B细胞和NEU细胞各自VCAM-1基因表达水平均明显上调。不同浓度葛根素干预后BEAS-2B细胞和NEU细胞的VCAM-1基因表达显著下调,BEAS-2B细胞和NEU细胞表面VCAM-1蛋白质相应减少。结论葛根素可有效抑制BEAS-2B与NEU共同培养诱导的BEAS-2B细胞和NEU细胞中VCAM-1基因及降低其细胞表面蛋白质平均荧光强度。

Objective To investigate the inhibiting effect of puerarin on VCAM-1 expression in co-culture of bronchial epithelial cells（BEAS-2B） and neutrophils（NEU） in vitro.Methods The mean fluorescence intensity of VCAM-1 on BEAS-2B and neutrophils was examined by flow cytometry,and the gene expression of VCAM-1 in the above cell lines was detemined by Real-Time PCR.Results The mean fluorescence intensity（MFI）of VCAM-1 of human bronchial epithelial cells co-cultured with human neutrophisl was significantly increased compared with human epithelial cells cultured alone.The MFI of VCAM-1 of human neutrophil cells co-cultured with human bronchial epithelial cells was significantly increased compared with human neutrophil cells cultured alone.The mRNA expression of VCAM-1 in human bronchial epithelial cells and neutrophil cells was significantly up-regulated in co-cultured systems.Incubated with puerarin,the MFI of VCAM-1 and the gene expression of VCAM-1 in human bronchial epithelial cells and neutrophil cells were significantly decreased.Conclusion Puerarin inhibits the mRNA expression and protein of VCAM-1 and the correspondent cell surface protein level in co-culture of bronchial epithelial cells and neutrophils.