幽门螺杆菌通过其优势蛋白组分活化肥大细胞的致病机制研究

Study on the pathological mechanism of Helicobacter pylori via its dominant protein components activating mast cells

目的探讨幽门螺杆菌(Helicobacter pylori,Hp)对人肥大细胞的活化作用,筛选出作用最强组分并评价其潜在活化效力。方法将3种Hp全菌制剂及其30种蛋白组分(Protein components,PCs)与LAD 2肥大细胞共培养,检测其对LAD 2肥大细胞的潜在活化作用,并筛出效力最强的PC;用液相色谱-质谱串联分析方法检测优势性PC成分,用ELISA法检测其激活LAD 2肥大细胞释放炎性介质的水平。结果3种Hp全菌制剂均有效活化LAD 2肥大细胞(P<0.05),并呈浓度依赖性;PC17具有最强的活化能力,能有效激活LAD 2肥大细胞释放组胺、TNF-α、IL-3、IFN-γ、LTB4(P<0.05),并呈浓度和时间依赖性;液相色谱-质谱分析和数据库比对揭示,PC17主要由23种蛋白组成,分子量为21~35 kDa。结论PC17是Hp中最强的肥大细胞活化成分,其可能通过促肥大细胞释放组胺、TNF-α、IL-3、IFN-γ、LTB4,参与Hp感染相关性疾病的发病。

The effect of Helicobacter pylori(Hp)on human mast cell remains unclear.Our research aimed to screen out the most efficient protein component and investigate its specific mast cell activation effects.After incubated with three kinds of Hp bacteria preparations and its 30 kinds of protein components(PCs),LAD 2 mast cell activation was assessed and the most efficient PC was screened out.Then,the specific protein ingredients and activation effects of the prominent PC were detected by liquid chromatography-mass spectrometry(LC-MS/MS)and ELISA assay respectively.The results showed that the three kinds of Hp preparations can activate LAD 2 mast cell in a dose-dependent manner(P<0.05)and PC17 was its most powerful PCs,which can stimulate the release of histamine,TNF-α,IL-3,IFN-γ,and LTB4 by LAD 2 mast cells in a dose-dependent or time-dependent manner.Twenty-three kinds of proteins were confirmed in PC17 by LC-MS/MS,and their molecular weights were ranged from 21 to 35 kDa.In conclusion,PC17 was the most powerful protein component in Hp with the mast cell activation ability,and can activate mast cells to secrete histamine,TNF-α,IL-3,IFN-γand LTB4 to participation in the pathogenesis of Hp-related diseases.