掌叶半夏毒针晶及凝集素蛋白的致炎作用与巨噬细胞的相关性研究  

Research on the relationship between raphides and lectin from Pinellia pedatisecta induce inflammatory and macrophage

作　　者：潘耀宗[1] 郁红礼[1,2,3] 吴皓[1,2,3] 陈叶青[1] 王奎龙[1] 

机构地区：[1]南京中医药大学药学院,南京210023 [2]江苏省中药炮制重点实验室,南京210023 [3]国家教育部中药炮制规范化及标准化工程研究中心,南京210023

出　　处：《中华中医药杂志》2014年第5期1397-1401,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金资助项目(No.30973939;No.81173549);教育部博士点新教师基金(No.20113237120010);南京中医药大学中药学江苏省优势学科开放课题(No.2011ZYX2-012)~~

摘　　要：目的:探索掌叶半夏毒针晶及凝集素蛋白诱导炎症与巨噬细胞的相关性。方法:自掌叶半夏块茎中提取分离纯化掌叶半夏毒针晶与凝集素蛋白;采用SDS-PAGE凝胶电泳联合质谱分析鉴定凝集素蛋白;采用大鼠腹腔巨噬细胞体外培养模型,以TNF-α和IL-1β为指标,研究掌叶半夏毒针晶及凝集素蛋白刺激巨噬细胞释放炎症因子的影响;采用扫描电镜法观察经掌叶半夏毒针晶和凝集素蛋白刺激后巨噬细胞的形态变化;采用巨噬细胞-中性粒细胞体外共培养迁移模型,考察掌叶半夏毒针晶和凝集素蛋白分别刺激巨噬细胞对中性粒细胞迁移的影响。结果:掌叶半夏毒针晶和凝集素蛋白刺激巨噬细胞后,巨噬细胞体积增大,伪足数量增加,细胞膜逐渐破裂。且两者体外均能显著诱导中性粒细胞迁移,其促迁移聚集作用与巨噬细胞释放炎症因子相关。结论:掌叶半夏毒针晶及凝集素蛋白诱导炎症与巨噬细胞介导密切相关。Objective： To Explore the relationship between the inflammation induced by raphides and lectin from Pinellia pedatisecta and macrophages. Methods： Raphides and lectin from Pinellia pedatisecta tuber were isolated and purified; Lectin was identified by using SDS-PAGE combined MS; Using TNF-a and IL-113 as indicators, the dose-response curve of inflammatory cytokines released by macrophages was studied; The morphological changes of macrophages stimulated by raphides and lectin from Pinellia pedatisecta were observed by using scanning electron microscope; Using the model of macrophages-neutrophil migration coculture, the inflections of neutrophil migration induced by raphides and lectin from Pinellia pedatisecta through stimulating macrophages was studied. Results： The volume of macrophages became bigger after raphides and lectin from Pinellia pedatisect stimulate macrophages, and the number of pseudopodium increased, and then the cell membrane ruptured gradually. What＇s more, raphides and lectin from Pinellia pedatisect could both induce neutrophil migration significantly in vitro, in addition, their effects of promoting migration and collection were due to inflammatory cytokines released by macrophages. Conclusion： The inflammation induced by raphides and lectin from Pinellia pedatisect is closely related to the mediation of macrophages.

关 键 词：掌叶半夏毒针晶 掌叶半夏凝集素蛋白 巨噬细胞 扫描电镜 体外中性粒细胞趋化 炎症因子 

分 类 号：R285[医药卫生—中药学]