葛根素对支气管上皮细胞和中性粒细胞共培养体系中黏附分子表达的影响  

Effect of puerarin on expression of adhesive molecule in co-culture of bronchial epithelial cells with neutrophils

作　　者：邵玲俐[1] 兰晓梅[2] 王成彬[2] 吕建新[1] 

机构地区：[1]温州医学院检验医学院、生命科学学院,浙江温州325035 [2]解放军总医院临床检验科,北京100853

出　　处：《解放军医学杂志》2012年第4期308-311,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(30873419)~~

摘　　要：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)的相互作用进而调控气道炎症反应的机制。方法实验分为BEAS-2B单独培养组、NEU单独培养组、BEAS-2B+NEU共培养组、BEAS-2B+NEU+50μg/ml葛根素共培养组、BEAS-2B+NEU+100μg/ml葛根素共培养组、BEAS-2B+NEU+200μg/ml葛根素共培养组,应用流式细胞术检测BEAS-2B和NEU两种细胞中胞间黏附分子ICAM-1、ICAM-3蛋白的表达,Real-time PCR检测两种细胞中ICAM-1、ICAM-3 mRNA的表达。结果与单独培养组相比,BEAS-2B+NEU共培养组中BEAS-2B细胞ICAM-1、ICAM-3及NEU细胞ICAM-1蛋白和mRNA表达均明显增强(P<0.05,P<0.01),而NEU细胞ICAM-3蛋白和mRNA表达无明显改变(P>0.05)。在BEAS-2B+NEU共培养体系中分别加入50、100、200μg/ml葛根素干预后,流式细胞术检测显示仅葛根素浓度为200μg/ml时BEAS-2B细胞ICAM-1蛋白表达显著低于BEAS-2B+NEU共培养组(P<0.05),余与BEAS-2B+NEU共培养组比较差异均无统计学意义,而Real-time PCR检测显示,除NEU细胞的ICAM-3 mRNA与BEAS-2B+NEU共培养组比较差异无统计学意义外,余均显著低于BEAS-2B+NEU共培养组(P<0.05,P<0.01)。结论葛根素可有效抑制BEAS-2B与NEU共同培养诱导的BEAS-2B及NEU细胞ICAM-1、ICAM-3基因和蛋白的表达。Objective To investigate the mechanism of puerarin-induced inhibition of the interaction between bronchial epithelial cells（BEAS-2B） and neutrophils（NEU）,which further regulates airway inflammation.Methods BEAS-2B cells and neutrophils were cultured alone,co-cultured,and co-cultured with different concentrations of puerarin（50μg/ml,100μg/ml,200μg/ml） for a pre-determined time.The Mean fluorescence intensity of intercellular adhesion molecule-1（ICAM-1） and ICAM-3 on BEAS-2B cells and neutrophils were determined by flow cytometry respectively,and the gene expressions of ICAM-1 and ICAM-3 in the two cell lines were determined by Real-time PCR.Results The protein expression of both ICAM-1 and ICAM-3 on BEAS-2B cells co-cultured with neutrophils were significantly higher than those cultured alone（P0.001,P0.05）.The protein expression of ICAM-1 on neutrophils co-cultured with BEAS-2B cells were also significantly higher than those cultured alone（P0.001）.There was no significant change of ICAM-3 expression in neutrophils before and after the co-culture.Compared with BEAS-2B cells cultured alone,the gene expression of both ICAM-1 and ICAM-3 were significantly up-regulated in those cells after co-cultured（P0.001,P0.01）.The gene expression of ICAM-1 in neutrophils was also significantly up-regulated after co-culture（P0.001）.With the intervention of different concentration of puerarin（50μg/ml,100μg/ml,200μg/ml）,flow cytometry showed that only the puerarin at 200μg/ml can inhibit the ICAM-1 expression of BEAS-2B cells co-cultured with neutrophils.Real-time PCR results showed no significant difference of ICAM-3 gene expression in human neutrophil cells between cultured alone and co-culture groups,while the ICAM-1 gene expression in human neutrophil cells and ICAM-1 and ICAM-3 gene expression in human bronchial epithelial cells when cultured alone were significantly lower than those in co-culture group（P0.001,P0.01）.Conclusion Puerarin inhibit both gene and protein expressions of I

关 键 词：上皮细胞 支气管 中性白细胞 细胞黏附分子 葛根素 

分 类 号：R364.5[医药卫生—病理学]