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作 者:庞伟[1,2] 兰晓梅[1] 邵玲俐[1] 王成彬[1]
机构地区:[1]解放军总医院临检科,北京医学硕士研究生100853 [2]武警医学院附属医院心内科,天津300162
出 处:《医学研究生学报》2011年第1期20-24,共5页Journal of Medical Postgraduates
基 金:国家自然科学基金(30873419)
摘 要: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.
关 键 词:葛根素 气道炎症 嗜中性粒细胞 单核细胞趋化蛋白-1
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