摘要
目的研究补体能否经由Fcα/μ受体(Fcα/μR)介导杀伤人肾小球系膜细胞。方法应用脂质体将带有Fcα/μRcDNA的质粒pcDNA3.1-Fcα/μR转染入人肾小球系膜细胞(NHMC),用Western blot检测Fcα/μR在NHMC细胞的表达,激光共聚焦显微术检测Fcα/μR在细胞膜的表达。用流式细胞术与激光共聚焦显微术检测所制备的IgM型免疫复合物(IgM-IC)与膜表达Fcα/μR的NHMC细胞的结合,用台盼蓝染色检测补体介导的细胞杀伤作用。结果成功制备了胞膜表达Fcα/μR的NHMC细胞,IgM与IgM-IC能经由Fcα/μR结合于NHMC细胞。在补体作用下,结合有IgM-IC的pcDNA3.1-Fcα/μR转染细胞死亡率显著高于对照组野生型细胞、空载质粒转染细胞及无IgM-IC结合的pcDNA3.1-Fcα/μR转染细胞(P<0.001)。结论胞膜表达Fcα/μR并结合有IgM-IC的人肾小球系膜细胞能为补体所杀伤。
Objective To study whether Fcα/μ receptor（Fcα/μR） can mediate complement killing of human glomerular mesangial cells.Methods Fcα/μR cDNA contained plasmid,pcDNA3.1-Fcα/μR was transfected into a human glomerular mesangial cell（NHMC）.Fcα/μR expression was detected by Western blot and laser scanning confocal microscopy.Binding of IgM-immune complexes（IgM-IC） to the Fcα/μR on cell membrane was detected by flowcytometry and laser scanning confocal microscopy.Killing of cells by complement was shown by Trypan blue exclusion assay.Results NHMC cells transfected with Fcα/μR could bind IgM and IgM-IC.After treatment with complement,added IgM-IC,the death rate of pcDNA3.1-Fcα/μR transfected cell was significant higher than the control groups of wild type cell,pcDNA3.1 transfected cell and the pcDNA3.1-Fcα/μR transfected cell without IgM-IC.Conclusion IgM-IC can bind to the Fcα/μR expressed NHMC cells and mediate complement killing of the cells.
出处
《基础医学与临床》
CSCD
北大核心
2010年第5期471-475,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(30972717)
