CAJ | 학술논문

目的探讨使用腺伴随病毒（AAV）载体介导编码多巴胺合成相关的三种生物酶基因的共转移。方法使用AAV载体将多巴胺生物合成相关的三种生物酶--酪氨酸氢化酶（TH）、芳香族氨基酸脱酶（AADC）、以及三磷酸鸟苷环水解酶Ⅰ（GGH）基因共转导入293细胞，以Western印迹分析检测TH、AADC、GCH的表达，以高效液相色谱法检测细胞内多巴胺水平。结果 TH、AADC、GCH的可在使用三种AAV载体（AAV-TH、AAV-AADC、AAV-GCH）转导的细胞中有效共表达，进一步分析发现，这种共表达可使该细胞有效地自发生成多巴胺。结论使用AAV载体进行TH、AADC、GCH三重基因共转导是有效的，这对帕金森病基因治疗的研究可能具有重要意义。
목적탐토사용선반수병독（AAV）재체개도편마다파알합성상관적삼충생물매기인적공전이。방법사용AAV재체장다파알생물합성상관적삼충생물매--락안산경화매（TH）、방향족안기산탈매（AADC）、이급삼린산조감배수해매Ⅰ（GGH）기인공전도입293세포，이Western인적분석검측TH、AADC、GCH적표체，이고효액상색보법검측세포내다파알수평。결과 TH、AADC、GCH적가재사용삼충AAV재체（AAV-TH、AAV-AADC、AAV-GCH）전도적세포중유효공표체，진일보분석발현，저충공표체가사해세포유효지자발생성다파알。결론사용AAV재체진행TH、AADC、GCH삼중기인공전도시유효적，저대파금삼병기인치료적연구가능구유중요의의。
Objective To explore triple gene transfer of dopamine synthetic enzymes with separate adeno-associated virus (AAV) vectors.Methods The genes for dopamine synthetic enzymes, tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC), and GTP cyclohydrolase Ⅰ (GCH, an enzyme critical for tetrahydrobiopterin synthesis) were cotransduced into 293 cells with separate AAV vectors. Expressions of TH, AADC and GCH were detected by Western blot analysis. Intracellular dopamine level was assayed by high-performance liquid chromatography.Results TH, AADC and GCH were effectively coexpressed in transduced cells with three separate AAV vectors, AAV-TH, AAV-AADC and AAV-GCH. Furthermore, the coexpression resulted in an effectively spontaneous dopamine production in cotransduced cells.Conclusion The triple transduction of TH, AADC and GCH genes with separate AAV vectors is effective, which might be important to gene therapy for Parkinson's disease.