Ma Xian-Cang, Chu Zheng, Zhang Xiao-Ling, Jiang Wen-Hui, Jia Min, Dang Yong-Hui, Gao Cheng-Ge
Department of Psychiatry, First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, China.
College of Medicine and Forensics, Xi'an Jiaotong University Health Science Center, Xi'an, China.
Neurochem Res. 2016 Jun;41(6):1375-80. doi: 10.1007/s11064-016-1841-0. Epub 2016 Feb 4.
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.
本研究旨在构建一种能在脑内表达NAP的重组腺相关病毒(rAAV),并通过鼻-脑途径,研究该病毒对接受旷场试验和强迫游泳试验的C57 BL/6小鼠是否能产生抗抑郁作用。用EcoRI酶切重组质粒pGEM-T Easy/NT4-NAP时,可获得297 bp的片段,经DNA测序证实NT4-NAP序列与设计序列一致。用EcoRI酶切重组质粒pSSCMV/NT4-NAP时,可见297 bp的片段。成纤维细胞的免疫组织化学染色显示,在NT4-NAP/AAV组中检测到了NAP的表达。末次给药后1天进行强迫游泳试验时,经鼻给予NT4-NAP/AAV显著缩短了不动时间。在旷场试验中,经鼻给予NT4-NAP/AAV并未改变小鼠的行为,因此强迫游泳试验中观察到的效果并非归因于非特异性的活动增加。结果表明,成功构建了一种能在细胞中表达NAP的重组AAV载体,NAP可能是抗抑郁治疗的潜在作用靶点。