蛇莓乙醇提取物的体外抗炎机制研究  

Anti-inflammatory Effect of Duchesnea indica(Andr) Focke in vitro

作　　者：庞然[1] 张淑玲[1] 赵雷[1] 刘双林[2] 董继华[3] 叶翩[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院感染科,武汉430022 [2]华中科技大学同济医学院附属同济医院泌尿外科,武汉430030 [3]华中科技大学同济医学院附属协和医院中心实验室,武汉430022

出　　处：《华中科技大学学报（医学版）》2009年第4期481-485,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的研究蛇莓乙醇提取物在体外的抗炎作用,初步探讨其抗炎机制。方法通过脂多糖(LPS)干预RAW264.7巨噬细胞系建立炎症细胞模型,ELISA法检测上清液中TNF-α、NO的分泌量,实时荧光定量RT-PCR法检测TNF-α、i NOS、血红素氧合酶-1(HO-1)的基因表达,Western blot法测定HO-1的蛋白表达量,免疫细胞化学法检验核因子-κB(NF-κB)的表达及分布变化。结果蛇莓提取物干预后细胞所分泌的炎症介质(TNF-α和NO)与炎症模型组相比均显著降低(均P<0.01),并存在剂量依赖关系;实时荧光定量RT-PCR结果显示蛇莓提取物干预后细胞TNF-α、i N-OS的mRNA表达水平显著降低,HO-1的mRNA表达水平明显升高,差异均有统计学意义,也存在剂量依赖关系;Western blot结果显示药物干预后HO-1的蛋白表达水平明显升高(P<0.01);免疫细胞化学法结果显示仅有LPS干预时,NF-κB表达增强且集中分布在细胞核,而药物干预时,NF-κB多分布在胞质。结论蛇莓提取物通过抑制NF-κB的激活,下调巨噬细胞表达炎症介质,同时促进HO-1的释放而发挥一定的抗炎作用。Objective To investigate the anti-inflammatory effect of Duchesnea indica（Andr） Focke in vitro.Methods Inflammatory cell model was established by LPS acting on the RAW 264.7 cell line.The secretion of TNF-α and NO in the supernatant was determined by ELISA.The expression of mRNA of TNF-α,iNOS and heme oxygenase 1（HO-1） was detected by real-time PCR,the expression of HO-1 protein by Western blot,and the distribution of nuclear factor-κB（NF-κB） by immunocytochemical method respectively.Results Immunocytochemistry revealed that cytoplasm stained to brown presented NF-κB inactivation after intervention of the Duchesnea indica（Andr） Focke,and cell nucleus stained to brown presented NF-κB activation after intervention of LPS.The expression of HO-1 mRNA and protein could be significantly enhanced by Duchesnea indica（Andr） Focke,while the expression of pro-inflammatory mediators TNF-α,NO（iNOS） protein and mRNA could be significantly reduced in a dose-dependent manner（P〈0.01）.Conclusion The Duchesnea indica（Andr） Focke can significantly resist inflammation by inhibiting the activation of NF-κB,regulating the expression and secretion of pro-inflammatory mediators catadromically in murine macrophages treated by LPS,and enhancing the expression of HO-1.

关 键 词：蛇莓 RAW264.7细胞系 核因子-ΚB 炎症介质 血红素氧合酶-1 

分 类 号：R282[医药卫生—中药学]