PMID- 28729555 OWN - NLM STAT- MEDLINE DCOM- 20190128 LR - 20190128 IS - 2045-2322 (Electronic) IS - 2045-2322 (Linking) VI - 7 IP - 1 DP - 2017 Jul 20 TI - Inhibition of phosphodiesterase-5 suppresses calcineurin/NFAT- mediated TRPC6 expression in pulmonary artery smooth muscle cells. PG - 6088 LID - 10.1038/s41598-017-06350-5 [doi] LID - 6088 AB - The up-regulation of transient receptor potential channel 6 (TRPC6) has been found to contribute to the proliferation of pulmonary artery smooth muscle cells (PASMCs), and inhibition of phosphodiesterase-5 (PDE5) has been shown to suppress TRPC6 expression in PASMCs. However, the molecular mechanisms underlying the up-regulation of TRPC6 expression and PDE5 modulation of TRPC6 expression in PASMCs remain largely unclear. The aim of this study is to address these issues. Endothelin-1 (ET-1) dose and time-dependently up-regulated TRPC6 expression in primary cultured rat PASMCs, and this was accompanied with the activation of calcineurin and subsequent translocation of NFATc4 to the nucleus. Further study indicated that inhibition of calcineurin by cyclosporine A or knockdown of NFATc4 using small interfering RNA suppressed ET-1-induced TRPC6 up-regulation. In addition, luciferase reporter assay showed that NFATc4 directly regulated the expression of TRPC6 in PASMCs. Inhibition of PDE5 by sildenafil suppressed ET-1-induced activation of calcineurin/NFATc4 signaling pathway and consequent TRPC6 up-regulation in PASMCs, while these inhibitory effects of sildenafil were abolished by PKG inhibitor Rp-8Br-cGMPs. Taken together, our study indicates that ET-1 stimulates TRPC6 expression by activation of calcineurin/NFATc4 signaling pathway, and inhibition of PDE5 suppresses calcineurin/NFATc4- mediated TRPC6 expression in PASMCs in a cGMP-PKG-dependent manner. FAU - Li, Shaojun AU - Li S AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Pan, Yilin AU - Pan Y AUID- ORCID: 0000-0002-4794-3191 AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Ke, Rui AU - Ke R AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Xie, Xinming AU - Xie X AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Zhai, Cui AU - Zhai C AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Shi, Wenhua AU - Shi W AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Wang, Jian AU - Wang J AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Yan, Xin AU - Yan X AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Chai, Limin AU - Chai L AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Wang, Qingting AU - Wang Q AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Zhang, Qianqian AU - Zhang Q AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Su, Xiaofan AU - Su X AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Yang, Lan AU - Yang L AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. FAU - Gao, Li AU - Gao L AD - Division of Allergy and Clinical Immunology, Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, MD, 21224, USA. FAU - Li, Manxiang AU - Li M AD - Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, P.R. China. manxiangli@hotmail.com. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20170720 PL - England TA - Sci Rep JT - Scientific reports JID - 101563288 RN - 0 (Endothelin-1) RN - 0 (NFATC Transcription Factors) RN - 0 (TRPC6 Cation Channel) RN - EC 3.1.4.35 (Cyclic Nucleotide Phosphodiesterases, Type 5) SB - IM MH - Animals MH - Cyclic Nucleotide Phosphodiesterases, Type 5/*metabolism MH - Endothelin-1/metabolism MH - Gene Expression Regulation MH - Male MH - Muscle, Smooth, Vascular/*cytology/*metabolism MH - Myocytes, Smooth Muscle/*metabolism MH - NFATC Transcription Factors/*metabolism MH - Pulmonary Artery/*cytology/*metabolism MH - Rats MH - Signal Transduction MH - TRPC6 Cation Channel/*genetics/metabolism PMC - PMC5519690 COIS- The authors declare that they have no competing interests. EDAT- 2017/07/22 06:00 MHDA- 2019/01/29 06:00 PMCR- 2017/07/20 CRDT- 2017/07/22 06:00 PHST- 2017/01/03 00:00 [received] PHST- 2017/06/13 00:00 [accepted] PHST- 2017/07/22 06:00 [entrez] PHST- 2017/07/22 06:00 [pubmed] PHST- 2019/01/29 06:00 [medline] PHST- 2017/07/20 00:00 [pmc-release] AID - 10.1038/s41598-017-06350-5 [pii] AID - 6350 [pii] AID - 10.1038/s41598-017-06350-5 [doi] PST - epublish SO - Sci Rep. 2017 Jul 20;7(1):6088. doi: 10.1038/s41598-017-06350-5.