Intranasal Delivery of Recombinant NT4-NAP/AAV Exerts Potential Antidepressant Effect
Intranasal Delivery of Recombinant NT4-NAP/AAV Exerts Potential Antidepressant Effect
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重组 NT4-NAP/AAV 鼻内递送发挥潜在的抗抑郁作用
DOI：
10.1007/s11064-016-1841-0
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发表时间：
2016-06-01
影响因子：
4.4
通讯作者：
Gao，Cheng-Ge
中科院分区：
文献类型：
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作者：
Ma，Xian-Cang;Chu，Zheng;Gao，Cheng-Ge
本研究旨在构建能在脑内表达NAP的重组腺相关病毒（rAAV），并通过旷场实验和强迫游泳实验观察其对C57 BL/6小鼠的抗抑郁作用。重组质粒pGEM-TEasy/NT 4-NAP经EcoRI酶切后，得到297 bp的片段，经DNA测序证实NT 4-NAP序列与设计序列一致。重组质粒pSSCMV/NT 4-NAP经EcoR Ⅰ酶切后，可扩增出297 bp的片段。成纤维细胞免疫组化染色显示，NT 4-NAP/AAV组有NAP表达。当在最后一次给药1天后进行FST时，鼻内递送NT 4-NAP/AAV显著减少了不动时间。在FST中观察到的效果不能归因于活性的非特异性增加，因为NT 4-NAP/AAV的鼻内递送在旷场试验期间不改变小鼠的行为。结果表明，成功构建了NAP的重组腺相关病毒载体，NAP有可能成为抗抑郁治疗的潜在靶点。
The present study was designed to construct a recombinant adeno-associated virus (rAAV) which can express NAP in the brain and examine whether this virus can produce antidepressant effects on C57 BL/6 mice that had been subjected to open field test and forced swimming test, via nose-to-brain pathway. When the recombinant plasmid pGEM-T Easy/NT4-NAP was digested by EcoRI, 297 bp fragments can be obtained and NT4-NAP sequence was consistent with the designed sequence confirmed by DNA sequencing. When the recombinant plasmid pSSCMV/NT4-NAP was digested by EcoRI, 297 bp fragments is visible. Immunohistochemical staining of fibroblasts revealed that expression of NAP was detected in NT4-NAP/AAV group. Intranasal delivery of NT4-NAP/AAV significantly reduced immobility time when the FST was performed after 1 day from the last administration. The effects observed in the FST could not be attributed to non-specific increases in activity since intranasal delivery of NT4-NAP/AAV did not alter the behavior of the mice during the open field test. The results indicated that a recombinant AAV vector which could express NAP in cells was successfully constructed and NAP may be a potential target for therapeutic action of antidepressant treatment.