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目的 探讨多巴胺合成酶基因的三重共转导能否进一步提高帕金森病大鼠纹状体多巴胺的生成。方法 以复合感染的方式，将分别构建编码酪氨酸羟化酶（ＴＨ）、芳香族氨基酸脱羧酶（ＡＡＤＣ）和三磷酸鸟苷环水解酶Ｉ（ＧＣＨ）基因的腺伴随病毒（ＡＡＶ）载体，通过立体定向法注射入帕金森病大鼠损毁侧纹状体。用高效液相色谱法进行纹状体ＢＨ４和多巴胺含量的测定；采用免疫组化方法确定ＴＨ、ＡＡＤＣ和ＧＣＨ在纹状体内的表达。结果 三重目的基因共转导可使大鼠纹状体ＢＨ４和多巴胺合成量的增加较ＴＨ和 ＡＡＤＣ三重基因转导进一步提高（Ｐ＜０．０５），组织学依据证明三种目的基因可在同一纹状体神经细胞内共同进行表达。结论 ＴＨ、ＡＡＤＣ和 ＧＣＨ三重基因共表达这一方法对探寻更为有效的帕金森病基因治疗策略可能具有重要的意义。
Objective To investigate whether triple co-transduction of dopamine synthase gene can further increase dopamine production in striatum of rats with Parkinson’s disease. Methods Adeno-associated virus (AAV) vector encoding tyrosine hydroxylase (TH), aromatic amino acid decarboxylase (AADC) and guanosine triphosphate cyclohydrolase I (GCH) The rats were injected into the Parkinson’s disease damaged lateral striatum by stereotactic method. The levels of striatum BH4 and dopamine were determined by high performance liquid chromatography. The expression of TH, AADC and GCH in the striatum was determined by immunohistochemistry. Results The co-transduction of triple gene could increase the synthesis of BH4 and dopamine in striatum more than that of TH and AADC (P <0.05), and the histological evidence showed that the three genes of interest could be in the same Striatum neurons co-expressed. Conclusion The co-expression of TH, AADC and GCH genes may play an important role in exploring a more effective strategy for gene therapy of Parkinson’s disease.