title: Prevention of vascular smooth muscle cell proliferation and injury-induced neointimal hyperplasia by CREB-mediated p21 induction: An insight from a plant polyphenol.
abstract: Cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/cAMP response element (CRE)-binding protein (CREB) signaling cascade negatively regulates platelet-derived growth factor BB (PDGF-BB)-induced smooth muscle cell (SMC) proliferation, which is a critical event in the initiation and development of restenosis and atherosclerotic lesions. Salvianolic acid A (SAA) is one of the most abundant polyphenols extracted from salvia. The aim of this study is to investigate whether SAA exerts an action on PDGF-BB-induced proliferation via cAMP/PKA/CREB mechanism. SAA blunts PDGF-BB-induced human umbilical artery smooth muscle cell (hUASMC) proliferation via p21 induction, as evidenced by its increased mRNA and protein expression levels. The SAA-induced upregulation of p21 involves the cAMP/PKA signaling pathway; a cAMP analog mimicked the effects of SAA and a specific cAMP/PKA inhibitor opposed these effects. SAA also activated CREB, including phosphorylation at Ser133, and induced its nuclear translocation. Deletion and mutational analysis of p21 promoters, co-immunoprecipitation, and western blot analysis showed that CRE is essential for SAA-induced p21 protein expression. Transfection of dominant-negative CREB (mutated Ser133) plasmids into hUASMCs attenuated SAA-stimulated p21 expression. SAA upregulated p21 expression and activated CREB in the neointima of balloon-injured arteries in vivo. Our results indicate that SAA promotes p21 expression in SMCs through the cAMP/PKA/CREB signaling cascade in vitro and prevents injury-induced neointimal hyperplasia.
authors.affiliation: State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, People's Republic of China; Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050, People's Republic of China.
authors.forename: Lan
authors.initials: L
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, People's Republic of China; Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050, People's Republic of China.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, People's Republic of China; Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050, People's Republic of China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Pharmacology, Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, People's Republic of China.
authors.forename: Weiku
authors.initials: W
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, People's Republic of China; Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050, People's Republic of China.
authors.forename: GuoRong
authors.initials: G
authors.lastname: He
authors.affiliation: State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, People's Republic of China; Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050, People's Republic of China.
authors.forename: Shengqian
authors.initials: S
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, People's Republic of China; Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050, People's Republic of China.
authors.forename: Junke
authors.initials: J
authors.lastname: Song
authors.affiliation: State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, People's Republic of China; Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050, People's Republic of China. Electronic address: dugh@imm.ac.cn.
authors.forename: Guanhua
authors.initials: G
authors.lastname: Du
mesh_terms: D000475:Alkenes / Q000494:pharmacology*; D000818:Animals; D020212:Carotid Artery Injuries / Q000473:pathology / Q000517:prevention & control; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000096:biosynthesis*; D006801:Humans; D006965:Hyperplasia; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000473:pathology; D059808:Polyphenols / Q000494:pharmacology*; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D017539:Tunica Intima / Q000187:drug effects* / Q000473:pathology
chemical_list: D000475:Alkenes; D017362:Cyclic AMP Response Element-Binding Protein; D050759:Cyclin-Dependent Kinase Inhibitor p21; D059808:Polyphenols; C568740:salvianolic acid; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases
keywords: 4′,6-Diamidino-2-phenylindole·2HCl (PubChem CID: 160166); 5-Bromo-2′-deoxyuridine (PubChem CID: 6035); 8-Bromoadenosine 3′,5′-cyclic monophosphate (PubChem CID: 32014); Cell proliferation; Cyclin-dependent kinase inhibitor p21; Polyphenols; Rp-Adenosine 3′,5′-cyclic monophosphorothioate triethylammonium (PubChem CID: 16760256); Salvianolic acid A (SAA, PubChem CID: 5281793); Vascular remodeling; cAMP response element-binding protein (CREB)