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The Expression of TRPC1/4 and Its Relationship with Proliferation and Apoptosis of Pulmonary Artery Smooth Muscle Cells under Hypoxic and Hypercapnic Conditions
Zheng Mengxiao1,2, Zhao Meiping1, Zhang Congcong1, Jia Xuguang3, Wu Yiming4, Chen Xiwen5, Wang Wantie1*
1Department of Pathophysiology, Wenzhou Medical University, Wenzhou 325035, China;
2School of Medicine, Yellow River Technical College, Zhengzhou 450063, China;
3Department of Internal Medicine, Yibin Health School, Yibin 644000, China;
4Division of Cardiovascular Medicine University of Iowa Carver College of Medicine, Iowa City 52242, USA;
5Experimental Animal Center, Wenzhou Medical University, Wenzhou 325035, China
2School of Medicine, Yellow River Technical College, Zhengzhou 450063, China;
3Department of Internal Medicine, Yibin Health School, Yibin 644000, China;
4Division of Cardiovascular Medicine University of Iowa Carver College of Medicine, Iowa City 52242, USA;
5Experimental Animal Center, Wenzhou Medical University, Wenzhou 325035, China
Abstract: This work was aimed to study the relationship between TRPC1/4 and pulmonary artery smooth muscle cells (PASMCs) proliferation and apoptosis under hypoxic and hypercapnic conditions. Cellular purity was assessed by immunofluorescence staining for α-SMA under fluorescence microscopy. PASMCs were divided into 5 groups randomly: normoxic group (N)， hypoxic and hypercapnic group (H)， DMSO group (D)， TRPC1/4 inhibitor SKF96365 group (S) and TRPC1/4 activator CPA group (C). N group was incubated under normoxia (5% CO2，21% O2， 37 °C) for 24 h， and the others were incubated under hypoxic and hypercapnic (6% CO2， 5% O2， 37 °C) atmosphere for 24 h. TRPC1/4 mRNA levels were detected by reverse transcription polymerase chain reaction (RTPCR).TRPC1/4 protein levels were detected by Western blot. The proliferation assay of PASMCs was performed by CCK-8 kit. The apoptosis of PASMCs was detected using the terminal deoxyribonucleotide transferase-mediated dUTP nick end-labeling (TUNEL) assay. [Ca2+]i was measured in PASMCs using fura 2-AM fluorescence. The results showed that the expression of TRPC1/4 mRNA and proteins and [Ca2+]i were upregulated under hypoxic and hypercapnic conditions. Hypoxia and hypercapnia promoted PASMCs proliferation and inhibited apoptosis.TRPC1/4 inhibitor SKF96365 reversed the effect of hypoxia and hypercapnia. CPA increased TRPC1 mRNA and protein levels， but neither TRPC4. The levels of TRPC1/4 mRNA and proteins and [Ca2+]i were upregulated under hypoxic and hypercapnic conditions. TRPC1 has a relationship with PASMCs proliferation and apoptosis.
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