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Role of TRPC6 in pulmonary artery smooth muscle cells proliferation and apoptosis under hypoxia and hypercapnia

JIA Xu-Guang, ZHENG Meng-Xiao, ZHANG Jing-Jing, ZHANG Cong-Cong, ZHAO Mei-Ping, WU Yi-Ming, CHEN Xi-Wen, WANG Wan-Tie

Department of Internal Medicine, Yibin Health School, Yibin 644000, China； School of Medicine, Huanghe Science and Technology College, Zhengzhou 450063, China； Department of Pathophysiology, Wenzhou Medical University, Wenzhou 325035, China； Division of Cardiovascular Medicine, University of Iowa Carver College of Medicine, Iowa City 52242, USA； Experimental Animal Center, Wenzhou Medical University, Wenzhou 325035, China

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+]i in the PASMCs was measured using Fura 2-AM fluorescence. The results showed that the expressions of TRPC6 mRNA and protein, and [Ca2+]i were 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.

Key words: pulmonary artery smooth muscle cells; pulmonary hypertension ; canonical transient receptor potential ; hypoxia and hypercapnia ; calcium

Received: 2016-04-17　　Accepted: 2016-10-28

Corresponding author: 王万铁　　E-mail: wwt@wmu.edu.cn

Citing This Article：

JIA Xu-Guang, ZHENG Meng-Xiao, ZHANG Jing-Jing, ZHANG Cong-Cong, ZHAO Mei-Ping, WU Yi-Ming, CHEN Xi-Wen, WANG Wan-Tie. Role of TRPC6 in pulmonary artery smooth muscle cells proliferation and apoptosis under hypoxia and hypercapnia. Acta Physiol Sin 2017; 69 (1): 47-54 (in Chinese with English abstract).