title: [Role of TRPC6 in pulmonary artery smooth muscle cells proliferation and apoptosis under hypoxia and hypercapnia].
abstract: The present study was to investigate the role of TRPC6 in pulmonary artery smooth muscle cells (PASMCs) proliferation and apoptosis under hypoxia and hypercapnia. PASMCs were isolated from chloral hydrate-anesthetized male Sprague-Dawley (SD) rats. Cellular purity was assessed by immunofluorescence staining for smooth muscle α-actin under fluorescence microscopy. Passage 4-6 PASMCs were starved for 24 h in serum-free DMEM and divided into 5 groups randomly: normoxia, hypoxia and hypercapnia, DMSO, TRPC6 inhibitor SKF-96365 and TRPC6 activator OAG groups. The normoxic group was incubated under normoxia (5% CO2, 21% O2, 37 °C) for 24 h, and the others were incubated with corresponding drugs under hypoxic and hypercapnic (6% CO2, 5% O2, 37 °C) atmosphere for 24 h. TRPC6 mRNA was detected by reverse transcription-PCR. TRPC6 protein was detected by Western blotting. The proliferation of PASMCs was performed by CCK-8 kit. Apoptosis of the PASMCs was detected using TUNEL assay. The [Ca2+]iin the PASMCs was measured using Fura 2-AM fluorescence. The results showed that the expressions of TRPC6 mRNA and protein, and [Ca2+]iwere upregulated under hypoxic and hypercapnic conditions. Hypoxia and hypercapnia promoted cellular proliferation and inhibited apoptosis in the PASMCs. OAG enhanced the above-mentioned effects of hypoxia and hypercapnia, whereas SKF-96365 reversed these effects. These results suggest that TRPC6 may play a role in PASMCs proliferation and apoptosis under hypoxia and hypercapnia by regulating [Ca2+]i.
authors.affiliation: Department of Internal Medicine, Yibin Health School, Yibin 644000, China.
authors.forename: Xu-Guang
authors.initials: XG
authors.lastname: Jia
authors.affiliation: School of Medicine, Huanghe Science and Technology College, Zhengzhou 450063, China.
authors.forename: Meng-Xiao
authors.initials: MX
authors.lastname: Zheng
authors.affiliation: Department of Pathophysiology, Wenzhou Medical University, Wenzhou 325035, China.
authors.forename: Jing-Jing
authors.initials: JJ
authors.lastname: Zhang
authors.affiliation: Department of Pathophysiology, Wenzhou Medical University, Wenzhou 325035, China.
authors.forename: Cong-Cong
authors.initials: CC
authors.lastname: Zhang
authors.affiliation: Department of Pathophysiology, Wenzhou Medical University, Wenzhou 325035, China.
authors.forename: Mei-Ping
authors.initials: MP
authors.lastname: Zhao
authors.affiliation: Division of Cardiovascular Medicine, University of Iowa Carver College of Medicine, Iowa City 52242, USA.
authors.forename: Yi-Ming
authors.initials: YM
authors.lastname: Wu
authors.affiliation: Experimental Animal Center, Wenzhou Medical University, Wenzhou 325035, China.
authors.forename: Xi-Wen
authors.initials: XW
authors.lastname: Chen
authors.affiliation: Department of Pathophysiology, Wenzhou Medical University, Wenzhou 325035, China. wwt@wmu.edu.cn.
authors.forename: Wan-Tie
authors.initials: WT
authors.lastname: Wang
mesh_terms: D000199:Actins; D000818:Animals; D017209:Apoptosis; D002118:Calcium / Q000378:metabolism; D015687:Cell Hypoxia; D049109:Cell Proliferation; D002478:Cells, Cultured; D006935:Hypercapnia / Q000503:physiopathology*; D007093:Imidazoles; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D011651:Pulmonary Artery / Q000166:cytology; D051381:Rats; D017207:Rats, Sprague-Dawley; D050052:TRPC Cation Channels / Q000378:metabolism*
chemical_list: D000199:Actins; D007093:Imidazoles; D050052:TRPC Cation Channels; C492145:Trpc6 protein, rat; C063159:1-(2-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenylethyl)-1H-imidazole; D002118:Calcium