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Seven days after the CCAO procedure, the authors performed encephalo-myo-synangiosis (EMS) and injected plasmid(s) into each rat's temporal muscle. Rats were divided into 4 groups based on which plasmid was received (i.e., LacZ group, VEGF group, apelin group, and VEGF+apelin group). Protein levels in the cortex and attached muscle were assessed with enzyme-linked immunosorbent assay (ELISA) on Day 7 after EMS, while immunofluorescent analysis of cortical vessels was performed on Day 14 after EMS.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>The total number of blood vessels in the cortex on Day 14 after EMS was significantly larger in the VEGF group and the VEGF+apelin group than in the LacZ group (p &lt; 0.05, respectively). Larger vessels appeared in the VEGF+apelin group than in the other groups (p &lt; 0.05, respectively). Apelin protein on Day 7 after EMS was not detected in the cortex for any of the groups. In the attached muscle, apelin protein was detected only in the apelin group and the VEGF+apelin group. Immunofluorescent analysis revealed that apelin and its receptor, APJ, were expressed on endothelial cells (ECs) 7 days after the CCAO.<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSIONS<\/jats:title><jats:p>Combined gene therapy (VEGF plus apelin) during EMS in a chronic cerebral hypoperfusion model can enhance angiogenesis in rats. This treatment has the potential to be a feasible option in a clinical setting for patients with moyamoya disease.<\/jats:p><\/jats:sec>","DOI":"10.3171\/2016.8.jns16366","type":"journal-article","created":{"date-parts":[[2016,12,23]],"date-time":"2016-12-23T15:30:42Z","timestamp":1482507042000},"page":"679-686","source":"Crossref","is-referenced-by-count":14,"title":["Combined gene therapy with vascular endothelial growth factor plus apelin in a chronic cerebral hypoperfusion model in rats"],"prefix":"10.3171","volume":"127","author":[{"given":"Masafumi","family":"Hiramatsu","sequence":"first","affiliation":[{"name":"Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomohito","family":"Hishikawa","sequence":"additional","affiliation":[{"name":"Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Koji","family":"Tokunaga","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Okayama City Hospital, Okayama; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroyasu","family":"Kidoya","sequence":"additional","affiliation":[{"name":"Department of Signal Transduction, Research Institute for Microbial Diseases, Osaka University, Suita, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shingo","family":"Nishihiro","sequence":"additional","affiliation":[{"name":"Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Haruma","sequence":"additional","affiliation":[{"name":"Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomohisa","family":"Shimizu","sequence":"additional","affiliation":[{"name":"Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuji","family":"Takasugi","sequence":"additional","affiliation":[{"name":"Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yukei","family":"Shinji","sequence":"additional","affiliation":[{"name":"Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenji","family":"Sugiu","sequence":"additional","affiliation":[{"name":"Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nobuyuki","family":"Takakura","sequence":"additional","affiliation":[{"name":"Department of Signal Transduction, Research Institute for Microbial Diseases, Osaka University, Suita, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isao","family":"Date","sequence":"additional","affiliation":[{"name":"Department of Neurological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1754","reference":[{"key":"b1-jns16366","series-title":"Neurosci Lett","doi-asserted-by":"publisher","first-page":"118","DOI":"10.1016\/j.neulet.2006.09.039","article-title":"Induced angiogenesis under cerebral ischemia by cyclooxygenase 2 and hypoxia-inducible factor naked DNA in a rat indirect-bypass model","volume":"409","author":"Anan M,","year":"2006","unstructured":"Anan M, Abe T, Matsuda T, Ishii K, Kamida T, Fujiki M, : Induced angiogenesis under cerebral ischemia by cyclooxygenase 2 and hypoxia-inducible factor naked DNA in a rat indirect-bypass model. 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