title: TRPC3-mediated Ca(2+) entry contributes to mouse airway smooth muscle cell proliferation induced by lipopolysaccharide.
abstract: Airway remodeling is a histopathological hallmark of chronic respiratory diseases that includes airway smooth muscle cell (ASMC) proliferation. Canonical transient receptor potential channel-3 (TRPC3)-encoded nonselective cation channels (NSCCs) are important native constitutively active channels that play significant roles in physiological and pathological conditions in ASMCs. Lipopolysaccharides (LPSs), known as lipoglycans and endotoxin, have been proven to be inducers of airway remodeling, though the mechanisms remain unclear. We hypothesized that TRPC3 is important in LPS-induced airway remodeling by regulating ASMC proliferation. To test this hypothesis, mouse ASMCs were cultured with or without LPS for 48h. Cell viability, TRPC3 protein expression, NSCC currents and changes in intracellular calcium concentration ([Ca(2+)]i) were then analyzed using an MTT assay, western blotting, whole-cell patch clamp and calcium imaging, respectively. The results showed that LPS treatment significantly induced ASMC proliferation, up-regulation of TRPC3 protein expression and enhancement of NSCC currents, resting [Ca(2+)]i and ACh-elicited changes in [Ca(2+)]i. TRPC3 blocker Gd(3+), TRPC3 blocking antibody or TRPC3 gene silencing by siRNA significantly inhibited LPS-induced up-regulation of TRPC3 protein, enhancement of NSCC currents, resting [Ca(2+)]i and ACh-elicited changes in [Ca(2+)]i, eventually inhibiting LPS-induced ASMCproliferation. These results demonstrated that TRPC3-mediated Ca(2+) entry contributed to LPS-induced ASMC proliferation and identified TRPC3 as a possible key target in airway remodeling intervention.
authors.affiliation: Department of Phamacology, School of Basic Medicine, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Xiao-Xu
authors.initials: XX
authors.lastname: Chen
authors.affiliation: Center for Stem Cell Research and Application, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
authors.forename: Jia-Hua
authors.initials: JH
authors.lastname: Zhang
authors.affiliation: Department of Phamacology, School of Basic Medicine, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Bin-Hua
authors.initials: BH
authors.lastname: Pan
authors.affiliation: Department of Phamacology, School of Basic Medicine, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Hui-Li
authors.initials: HL
authors.lastname: Ren
authors.affiliation: Department of Phamacology, School of Basic Medicine, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Xiu-Ling
authors.initials: XL
authors.lastname: Feng
authors.affiliation: Department of Phamacology, School of Basic Medicine, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Jia-Ling
authors.initials: JL
authors.lastname: Wang
authors.affiliation: Department of Phamacology, School of Basic Medicine, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China; The Key Laboratory for Drug Target Researches and Pharmacodynamic Evaluation of Hubei Province, Wuhan 430030, China. Electronic address: xiaoj@hust.edu.cn.
authors.forename: Jun-Hua
authors.initials: JH
authors.lastname: Xiao
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D005260:Female; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism; D012137:Respiratory System / Q000187:drug effects* / Q000378:metabolism; D050052:TRPC Cation Channels / Q000378:metabolism*
chemical_list: D008070:Lipopolysaccharides; D050052:TRPC Cation Channels; C104603:TRPC3 cation channel; D002118:Calcium
keywords: Airway remodeling; Airway smooth muscle cell; Lipopolysaccharide; Nonselective cation channel; Proliferation; TRPC3; [Ca(2+)](i)