CAJ | 학술논문

目的：构建一种能稳定表达人水通道蛋白－9（ AQP9）的真核细胞载体，并检测视神经脊髓炎（NMO）患者血清中的AQP9抗体。方法将人AQP9基因克隆入pcDNA3．1＋质粒中构建AQP9－pcDNA3．1＋载体，并转染HEK－293T细胞中。 AQP9蛋白的表达通过RT－PCR和Western blot法验证。通过间接免疫荧光法检测NMO患者血清中的AQP9抗体。结果 AQP9的mRNA相对表达量明显高于空载体，且蛋白表达量在32 kD条带附近明显高于空载和293T细胞。在4例NMO患者血清中均检测到AQP9抗体，1例脑梗死和1例健康人中抗体均为阴性。结论这种转染表达AQP9的细胞模式相对简捷，可用来检测NMO患者中的AQP9抗体，对NMO研究有一定意义。
목적：구건일충능은정표체인수통도단백－9（ AQP9）적진핵세포재체，병검측시신경척수염（NMO）환자혈청중적AQP9항체。방법장인AQP9기인극륭입pcDNA3．1＋질립중구건AQP9－pcDNA3．1＋재체，병전염HEK－293T세포중。 AQP9단백적표체통과RT－PCR화Western blot법험증。통과간접면역형광법검측NMO환자혈청중적AQP9항체。결과 AQP9적mRNA상대표체량명현고우공재체，차단백표체량재32 kD조대부근명현고우공재화293T세포。재4례NMO환자혈청중균검측도AQP9항체，1례뇌경사화1례건강인중항체균위음성。결론저충전염표체AQP9적세포모식상대간첩，가용래검측NMO환자중적AQP9항체，대NMO연구유일정의의。
Objective To construct a eukaryotic aquaporin -9 (AQP9) expressing vector for AQP9 antibody de-tection in patients with neuromyelitis optica (NMO).Methods The recombinant vector was constructed by subcloning human AQP9 cDNA into pcDNA3.1+vector, with which human embryonic kidney (HEK) 293T cells was transfected to establish an AQP9 protein expressing cell line .The cell line expression AQP 9 was confirmed by reverse transcription poly-merase chain reaction ( RT-PCR) and Western blot analysis .Serum antibody was tested with cell -based assay by indi-rect immunofluorescence assay .Results The relative expression level of AQP 9 mRNA was significantly higher than the empty vector, and Western blotting analysis showed the expression content at 32 kD bands was significantly higher than the empty vector and 293T cells.The cell line expressing AQP9 was successfully constructed and confirmed by reverse tran-scription polymerase chain reaction ( RT-PCR) and Western blot analysis .The serum antibody was positive in all 4 pa-tients with NMO and negative in one with cerebral infarction and one normal adult .Conclusion The transfected cell mod-el is simple and advantageous in AQP 9 serum antibody detection for research on NMO .