内毒素对肥大细胞脱颗粒释放组胺和类胰蛋白酶的影响

Influence of endotoxin on mast cell degranulation releasing histamine and tryptase

目的通过体外分离培养大鼠腹腔肥大细胞(RPMC),进而探讨内毒素(endotoxin,ET)对RPMC脱颗粒释放组胺和类胰蛋白酶的影响。方法将悬浮培养的RPMC分为5组:①无ET处理组;②0.5μg/mlET处理组;③1μg/mlET处理组;④2μg/mlET处理组;⑤4μg/mlET处理组。37℃孵育2h,收集RPMC上清,荧光法测定RPMC培养上清中组胺含量;专性底物α-N-苯甲酰-DL-精氨酸-P-硝基苯胺(BAPNA)测定RPMC培养上清中类胰蛋白酶含量,分析不同浓度ET对RPMC释放组胺和类胰蛋白酶的影响。结果不同浓度ET处理组与无ET处理组相比RPMC脱颗粒现象明显;不同浓度ET处理组刺激RPMC释放组胺和类胰蛋白酶的量均高于无ET处理组(P<0.05),与ET剂量依赖性不明显(P>0.05),并且给予1μg/mlET处理时组胺和类胰蛋白酶的释放量最高。结论ET体外可刺激RPMC脱颗粒释放组胺和类胰蛋白酶,但与ET剂量依赖性不明显。

Objective To explore the influence of endotoxin （ET） on the release of histamine and tryptase from mast cell degranulation by culturing and isolating rat peritoneal mast cells （RPMCs） in vitro. Methods After collecting the supernatant of RPMCs treated with different concentrations of ET（0.5, 1, 2, 4 μg/ml, respectively）, the histamine （HA） concentrations and tryptase （TS） activities in supernatant were detected. Results Compared to the group without ET treatment, RPMCs showed a obvious degranulation in ET treated groups, and HA concentrations and TS activities in the supernatant of RPMCs increased significantly in ET treated groups （P〈 0.05）, but not dose-dependently increased （P 〉 0.05）. HA concentrations and TS activities in the supernatant of RPMCs were the highest in 1 μg/ml ET treated group. Condusion ET can stimulate RPMCs to release HA and TS by the degranulation in vitro, but not dose-dependently.