title: Vascular smooth muscle cell contractile protein expression is increased through protein kinase G-dependent and -independent pathways by glucose-6-phosphate dehydrogenase inhibition and deficiency.
abstract: Homeostatic control of vascular smooth muscle cell (VSMC) differentiation is critical for contractile activity and regulation of blood flow. Recently, we reported that precontracted blood vessels are relaxed and the phenotype of VSMC is regulated from a synthetic to contractile state by glucose-6-phosphate dehydrogenase (G6PD) inhibition. In the current study, we investigated whether the increase in the expression of VSMC contractile proteins by inhibition and knockdown of G6PD is mediated through a protein kinase G (PKG)-dependent pathway and whether it regulates blood pressure. We found that the expression of VSMC-restricted contractile proteins, myocardin (MYOCD), and miR-1 and miR-143 are increased by G6PD inhibition or knockdown. Importantly, RNA-sequence analysis of aortic tissue from G6PD-deficient mice revealed uniform increases in VSMC-restricted genes, particularly those regulated by the MYOCD-serum response factor (SRF) switch. Conversely, expression of Krüppel-like factor 4 (KLF4) is decreased by G6PD inhibition. Interestingly, the G6PD inhibition-induced expression of miR-1 and contractile proteins was blocked by Rp-β-phenyl-1,N2-etheno-8-bromo-guanosine-3',5'-cyclic monophosphorothioate, a PKG inhibitor. On the other hand, MYOCD and miR-143 levels are increased by G6PD inhibition through a PKG-independent manner. Furthermore, blood pressure was lower in the G6PD-deficient compared with wild-type mice. Therefore, our results suggest that the expression of VSMC contractile proteins induced by G6PD inhibition occurs via PKG1α-dependent and -independent pathways.
authors: Biochemistry & Molecular Biology, University of South Alabama, Mobile, Alabama; Pharmacology, New York Medical College, Valhalla, New York.
authors: Sukrutha
authors: S
authors: Chettimada
authors: Biochemistry & Molecular Biology, University of South Alabama, Mobile, Alabama; Pharmacology, New York Medical College, Valhalla, New York.
authors: Sachindra Raj
authors: SR
authors: Joshi
authors: Pharmacology, New York Medical College, Valhalla, New York.
authors: Vidhi
authors: V
authors: Dhagia
authors: Aab Cardiovascular Research Institute, University of Rochester School of Medicine and Dentistry, Rochester, New York; and.
authors: Alessandro
authors: A
authors: Aiezza
authors: Physiology, University of South Alabama, Mobile, Alabama.
authors: Thomas M
authors: TM
authors: Lincoln
authors: Biochemistry & Molecular Biology, University of South Alabama, Mobile, Alabama; Pharmacology, New York Medical College, Valhalla, New York.
authors: Rakhee
authors: R
authors: Gupte
authors: Aab Cardiovascular Research Institute, University of Rochester School of Medicine and Dentistry, Rochester, New York; and.
authors: Joseph M
authors: JM
authors: Miano
authors: Biochemistry & Molecular Biology, University of South Alabama, Mobile, Alabama; Pharmacology, New York Medical College, Valhalla, New York.
authors: Sachin A
authors: SA
authors: Gupte
mesh_terms: D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism*; D015153:Blotting, Western; D002417:Cattle; D002853:Chromatography, Liquid; D003285:Contractile Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D062965:Cyclic GMP-Dependent Protein Kinase Type I / Q000037:antagonists & inhibitors / Q000378:metabolism*; D017869:Cyclic GMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D055785:Gene Knockdown Techniques; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors* / Q000235:genetics; D047468:Immunoprecipitation; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000187:drug effects / Q000235:genetics; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism*; D009687:Nuclear Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D051381:Rats; D026362:Serum Response Factor / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D015534:Trans-Activators / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000099251:Myocardin
chemical_list: D003285:Contractile Proteins; D062965:Cyclic GMP-Dependent Protein Kinase Type I; D017869:Cyclic GMP-Dependent Protein Kinases; D005954:Glucosephosphate Dehydrogenase; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D035683:MicroRNAs; D009687:Nuclear Proteins; D026362:Serum Response Factor; D015534:Trans-Activators; D000099251:Myocardin; C000715468:Klf4 protein, mouse; C000715469:Klf4 protein, rat; C517731:Mirn1 microRNA, mouse; C532884:MIRN143 microRNA, mouse
keywords: NADPH redox; RNA-seq; epigenetics; metabolism; pentose phosphate pathway; protein kinase G; vascular smooth muscle phenotype