title: Intracellular calcium concentration of corpus cavernosum smooth muscle cells is decreased by the overexpression of PnNOS gene in adipose tissue-derived stem cells.
abstract: The study investigated the effects of adipose tissue-derived stem cells (ADSCs) modified with penile neuronal nitric oxide synthase (PnNOS) gene on intracellular calcium concentration in rat corpus cavernosum smooth muscle cells (CCSMCs). ADSCs and CCSMCs of Sprague-Dawley (SD) rats were isolated and cultured in vitro respectively. The rat PnNOS gene was transferred into the ADSCs mediated by a recombinant adenovirus vector. The expression of the PnNOS gene was detected. At the same time, the concentration of nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) was assayed. After coculturing with the CCSMCs of SD rats, which were isolated and expanded ex vivo, the cGMP and NO levels of ADSCs and CCSMCs were measured. Intracellular calcium concentration ([Ca(2+) ]i ) in rat CCSMCs was measured with Fluo-3/AM by flow cytometer after cocultured with ADSCs overexpressing PnNOS gene. The mRNA and protein expression of PnNOS gene mediated by recombinant adenovirus vector significantly overexpressed and lasted at least 2 weeks. Meanwhile, the concentration of NO and cGMP in ADSCs was greatly increased. The concentration of cGMP was significantly increased, and [Ca(2+) ]i was obviously decreased in CCSMCs compared with the control groups (P < 0.05) after cocultured with ADSCs for 3 days. These findings demonstrated that ADSCs overexpressing PnNOS gene might decrease [Ca(2+) ]i in CCSMCs by up-regulating NO-cGMP signalling pathway.
authors.affiliation: Department of Infertility and Sexual Medicine, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.|School of Information and Optoelectronic Science and Engineering, South China Normal University, Guangzhou Guangdong, China.
authors.forename: J
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Infertility and Sexual Medicine, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Hu
authors.affiliation: Center of Reproductive Medicine, The First People's Hospital of Yunnan Province, Kunming, China.|Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: L
authors.initials: L
authors.lastname: Zhuan
authors.affiliation: Department of Urology, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
authors.forename: H
authors.initials: H
authors.lastname: Xiao
authors.affiliation: Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: J
authors.initials: J
authors.lastname: Yang
mesh_terms: D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000032:analysis* / Q000378:metabolism; D006152:Cyclic GMP / Q000032:analysis; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D008297:Male; D039902:Multipotent Stem Cells / Q000737:chemistry / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000737:chemistry / Q000378:metabolism*; D052248:Nitric Oxide Synthase Type I / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010413:Penis / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology
chemical_list: D052248:Nitric Oxide Synthase Type I; D006152:Cyclic GMP; D002118:Calcium
keywords: Adipose tissue-derived stem cells; corpus cavernosum smooth muscle cells; gene therapy; intracellular calcium concentration; penile neuronal nitric oxide synthase gene