帕金森病大鼠三重目的基因共表达的长期行为改善

Long-term behavioral recovery in Parkinsonian rats by triple transduction with dopamine-biosynthesizing enzyme genes

目的 探讨多巴胺合成酶基因的三重共转导对帕金森病基因治疗的效果。方法 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 ,用复合感染的方式将AAV TH、AAV AADC及AAV GCH通过立体定向法注射入帕金森病大鼠损毁侧纹状体。采用免疫组化方法确定TH、AADC和GCH的表达 ;并连续观测基因转导后大鼠行为改善的情况长达 1年。结果 组织学依据显示TH、AADC和GCH基因均可在纹状体内长期有效而稳定的进行表达 ;三重基因转导可引起大鼠较TH和AADC共转导更为显著的行为改善(P <0 0 1)。结论 AAV载体介导的TH、AADC和GCH三重基因纹状体内共表达对于帕金森病大鼠的基因治疗更为有效。

Objective To observe the effect of gene therapy for Parkinson disease by triple transduction with dopamine-biosynthesizing enzyme genes. Methods We constructed high titer purified adeno-associated virus (AAV) vectors containing cDNAs encoding human tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC) and GTP cyclohydrolase I (GCH) respectively. AAV-TH, AAV-AADC and AAV-GCH were stereotaxically injected into the denervated striatum of Parkinsonian rat; the expression of TH, AADC or GCH was assayed by using immunohistochemical staining; the rotational behavior was assayed for 1 year.Results The immunohistochemical staining and behavioral testing showed that all of TH, AADC and GCH were expressed stably in the striatum, and the rats treated with AAV-TH, AAV-AADC and AAV-GCH showed more remarkable decrease in rotation rate persisted for at least 1 year, than those treated with AAV-TH and AAV-AADC(P<0.01). Conclusion AAV-mediated striatal transgene expression of TH, AADC and GCH is more effective for gene therapy on Parkinsonian rat.