葛根素对支气管上皮细胞和嗜中性粒细胞共培养体系单核细胞趋化蛋白-1表达的影响

Puerarin reduces the expression of MCP-1 in co-cultured bronchial epithelial cells and neutrophils

目的嗜中性粒细胞(Neutroph il,NEU)可介导支气管上皮细胞(BEAS-2B)的炎性反应,葛根素具有抑制炎性细胞趋化作用。文中探讨葛根素对BEAS-2B细胞和NEU共培养体系中BEAS-2B细胞分泌单核细胞趋化蛋白-1(Monocytechenoattractant prote in-1,MCP-1)的影响。方法应用ELISA方法测量BEAS-2B细胞和NEU细胞单独培养、联合培养、加葛根素干预培养体系不同培养时间MCP-1浓度,应用实时定量PCR方法检测上述不同培养体系培养12 h后BEAS-2B细胞和NEU细胞中MCP-1基因表达。结果 BEAS-2B细胞和NEU细胞联合培养上清液中MCP-1浓度显著高于2种细胞单独培养所分泌MCP-1,联合培养的BEAS-2B细胞MCP-1基因表达水平明显上调。经葛根素干预的细胞联合培养上清液MCP-1浓度显著低于未经葛根素干预的细胞联合培养体系MCP-1浓度,前者较后者基因表达水平下调。结论 NEU细胞与BEAS-2B细胞联合培养可上调BEAS-2B细胞中MCP-1基因表达和显著增加细胞培养上清液中MCP-1浓度;葛根素可有效抑制BEAS-2B细胞和NEU细胞共培养体系MCP-1分泌和下调BEAS-2B细胞中MCP-1基因表达。

Objective Neutrophils（NEU） mediate the inflammatory reaction of bronchial epithelial（BEAS-2B） cells,and puerarin could inhibit the chemotaxis of inflammatory cells.This study was to investigate the effect of puerarin on the expression of monocyte chenoattractant protein-1（MCP-1） in the co-culture of human bronchial epithelial cells and neutrophils.Methods We constructed a system of human bronchial epithelial cells co-cultured with human neutrophils,and divided the cells into six groups to be cultured singly（BEAS-2B and NEU groups）,or co-cultured（BEAS-2B＋NEU group）,or intervened with puerarin at 50,100 and 200 μg/ml（PUE50,PUE100,and PUE200 groups）.The concentration of MCP-1 in the cell supernatant of each group was detected by ELISA after cultured for 2,6,12 and 18 hours,and the expression of the MCP-1 gene determined by real-time PCR.Results In the BEAS-2B＋NEU group,the expression of the MCP-1 gene was significantly up-regulated and the MCP-1 concentration was remarkably higher than either the BEAS-2B or the NEU group.The PUE intervention groups showed obviously decreased expression of the MCP-1 gene and significantly lower concentration of MCP-1 than in the BEAS-2B＋NEU group.Conclusion The co-culture of NEU and BEAS-2B can up-regulate the expression of the MCP-1 gene in BEAS-2B cells and increase the concentration of MCP-1 in the cell supernatant.Puerarin can effectively down-regulate the expression of the MCP-1 gene in BEAS-2B cells and inhibit the secretion of MCP-1 in the co-cultured BEAS-2B cells and human neutrophils.