烙铁头蛇毒TM-N49诱导人肥大细胞释放组胺的作用  

Induction of histamine release from human mast cells by trimeresurus mucrosquamatus venom TM-N49

作　　者：莫雅贞[1] 何韶衡[1] 魏继福[1] 林梓霞[2] 傅意玲[1] 

机构地区：[1]汕头大学医学院变态反应学与炎症学研究所,广东汕头515031 [2]广东省汕头市中心医院

出　　处：《海南医学》2006年第9期140-143,共4页Hainan Medical Journal

基　　金：国家自然科学基金项目(No.30471601);广东省科技计划重点项目(No.2003B31502);广东省自然科学基金重点项目(No.04106122);李嘉诚基金资助(No.C0200001)

摘　　要：目的探索烙铁头蛇毒TM-N49对人肥大细胞释放组胺的诱导作用及其机制。方法用SP-SephadexC-25,肝素凝胶,Superdex75分子筛及HPLC纯化TM-N49。将手术切除的人肺、大肠和扁桃体组织在37℃条件下用Ⅰ型胶原酶和Ⅰ型透明质酸酶消化,经悬浮后的细胞均匀地加入含TM-N49、刺激剂或缓冲液的试管中,进行激发实验并用荧光显色法测量上清液的组胺水平。结果纯化后的TM-N49在SDS-PAGE上呈一条带。TM-N49能诱导人肺、大肠和扁桃体肥大细胞发生剂量依赖性组胺释放。浓度为0.3μg/ml时,TM-N49就能诱导大肠肥大细胞显著性的组胺释放,但对于人肺肥大细胞来说,诱导它释放相同量组胺所需最低TM-N49浓度为3μg/ml,而对于扁桃体肥大细胞,TM-N49浓度为30μg/ml时才能诱导它显著性的组胺释放。TM-N49诱导大肠和肺肥大细胞组胺的释放10min和8min时达高峰,而扁桃体肥大细胞的组胺释放则在16min时达高峰。人大肠、肺和扁桃体肥大细胞经代谢抑制剂和百日咳毒素预先处理后,浓度低于30μg/ml的TM-N49诱导其释放组胺的作用明显减弱,但当其浓度达到30μg/ml时,上述处理对TM-N49诱导人肥大细胞释放组胺的作用无影响。缺乏外源性钙、镁离子时,TM-N49诱导肥大细胞释放组胺的能力也明显减弱。结论TM-N49可能通过G蛋白偶联受体途径激活人肥大细胞或通过细胞毒等途径作用于肥大细胞,从而参与毒蛇咬伤人体后所发生的病理生理过程。AIM： To investigate the ability of trimeresurus mucrosquamatus venom TM-N49 to induce the histamine release from human mast cell and its potential mechanisms, Methods TM-N49 was purified from the snake venom by using SP-Sephadex C-2.5, heparin agarose, Superdex75 chromatography and HPLC. Mast cells were dispersed from human lung, colon and tonsil tissues after digestion with collaganase and hyaluronidase, The dispersed mast cells were then challenged with TM-N49, stimulus and control in LP4 tubes for 15min at 37℃. A glass fibre-based fluorometric assay was used to measure histamine in the supernatants of dispersed mast cell. Results TM-N49 induced a dose dependent release of histamine from human colon, lung and tonsil mast cells. As low as 0.3μg/ml of TM-N49 was able to stimulate significant histamine release from human colon mast cells, hut a minimum of 3 or 30μg/ml of TM-N49 was required to stimulate a similar level of histamine release from lung or tonsil mast cells, respectively. The release of histamine from colon and lung mast cells in response to TM-N49 was maximized at 10 and 8 min following addition of the stimulus. Tnis was quite different from the picture of the peak histamine release from tonsil mast cells, in which histamine release was maximized at 16 rain following addition of TM-N49. Pretreatment of cells with metabolic inhibitors and pertussis toxin reduced dramatically histamine release from human colon, lung and tonsil mast cells by TM-N49 when below 30μg/ml, while TM-N49 is over 30μg/mL this treatments had no effect on them. In no exogenous Ca^2＋ and Mg^2＋ experiments, the release of histamine induced by TM-N49 was significantly decreased. Condusions Trimeresurus mucrosquamatus venom TM-N49 can induce human mast cells to release histamine through a G-protein-related or a cytotoxic mechanism, which may contribute to the pathogenesis of venomous snake bite.

关 键 词：肥大细胞 烙铁头蛇毒TM—N49 组胺 

分 类 号：R-33[医药卫生]