整合素和钙通道在人与小鼠钩端螺旋体主要宿主细胞上分布的差异

Distribution diversity of integrins and calcium channels on major human and mouse host cells of Leptospira species

目的:了解人和小鼠钩端螺旋体(简称钩体)主要宿主细胞上整合素与钙通道表达情况及其差异。方法:采用免疫荧光和Western blot法,检测人单核细胞株THP-1、小鼠单核!巨噬样细胞株J774A.1、人脐静脉内皮细胞株HUVEC、小鼠血管内皮细胞株EOMA、人肝细胞株L-02、小鼠肝细胞株Hepa1-6、人肾小管上皮细胞株HEK-293、小鼠肾小球系膜上皮细胞株SV40-MES13、小鼠肾成纤维细胞株NIH/3T3、人及小鼠血小板的β1、β2、β3类整合素分子以及Cav3.1、Cav3.2、Cav3.3、Cav2.3电压门控型钙通道蛋白和P2X1、P2X2、P2X3、P2X4、P2X5、P2X6、P2X7受体门控型钙通道蛋白的分布差异。结果:整合素β1分子表达于J774A.1、THP-1、HUVEC、EOMA、L-02、Hepa1-6、HEK-239细胞以及人和小鼠血小板膜表面,β2分子表达于J774A.1、THP-1、HUVEC、EOMA和NIH-3T3细胞膜表面,β3分子表达于J774A.1、THP-1、HUVEC、EOMA、Hepa1-6、HEK-239、NIH-3T3细胞以及人和小鼠血小板膜表面。ATP依赖性受体门控型钙通道蛋白P2X1表达于人和小鼠血小板膜表面,P2X5表达于J774A.1、THP-1、L-02、Hepa1-6、HEK-239和HUVEC细胞膜表面,但未检测出上述细胞或血小板表达其它钙通道蛋白。结论:不同种类整合素分子和钙通道蛋白在钩体主要宿主细胞上表达明显不同,这可能与钩体对不同组织细胞致病性差异有关。

Objective： To determine the distribution of integrins and calcium channels on major human and mouse host cells of Leptospira species.Methods： The expression of β1,β2 and β3 integrins was detected with immunofluorescence assay on the surface of human monocyte line THP-1,mouse mononuclear-macrophage-like cell line J774A.1,human vascular endothelial cell line HUVEC,mouse vascular endothelial cell EOMA,human hepatocyte line L-02,mouse hepatocyte line Hepa1-6,human renal tubular epithelial cell line HEK-293,mouse glomerular membrane epithelial cell line SV40-MES13,mouse collagenoblast line NIH/3T3,human and mouse platelets.The distribution of voltage gate control calcium channels Cav3.1,Cav3.2,Cav3.3 and Cav2.3,and receptor gate calcium channels P 2X 1,P 2X 2,P 2X 3,P 2X 4,P 2X 5,P 2X 6and P 2X 7were determined with Western blot assay.Results： β1 integrin proteins were positively expressed on the membrane surface of J774A.1,THP-1,HUVEC,EOMA,L-02,Hepa1-6 and HEK-239 cells as well as human and mouse platelets.β2 integrin proteins were expressed on the membrane surface of J774A.1,THP-1,HUVEC,EOMA,and NIH/3T3 cells.β3 integrin proteins were expressed on the membrane surface of J774A.1,THP-1,HUVEC,EOMA,Hepa1-6,HEK-239 and NIH/3T3 cells as well as human and mouse platelets.P 2X 1receptor gate calcium channel was expressed on the membrane surface of human and mouse platelets,while P 2X 5receptor gate calcium channel was expressed on the membrane surface of J774A.1,THP-1,L-02,Hepa1-6,HEK-239 and HUVEC cells.However,the other calcium channels were not detected on the tested cell lines or platelets.Conclusion： There is a large distribution diversity of integrins and calcium channel proteins on the major human and mouse host cells of Leptospira species,which may be associated with the differences of leptospire-induced injury in different host cells.