title: NFkappaB activation is essential for miR-21 induction by TGFβ1 in high glucose conditions.
abstract: Transforming growth factor beta1 (TGFβ1) is a pleiotropic growth factor with a very broad spectrum of effects on wound healing. Chronic non-healing wounds such as diabetic foot ulcers express reduced levels of TGFβ1. On the other hand, our previous studies have shown that the microRNA miR-21 is differentially regulated in diabetic wounds and that it promotes migration of fibroblast cells. Although interplay between TGFβ1 and miR-21 are studied in relation to cancer, their interaction in the context of chronic wounds has not yet been investigated. In this study, we examined if TGFβ1 could stimulate miR-21 in fibroblasts that are subjected to high glucose environment. MiR-21 was, in fact, induced by TGFβ1 in high glucose conditions. The induction by TGFβ1 was dependent on NFκB activation and subsequent ROS generation. TGFβ1 was instrumental in degrading the NFκB inhibitor IκBα and facilitating the nuclear translocation of NFκB p65 subunit. EMSA studies showed enhanced DNA binding activity of NFκB in the presence of TGFβ1. ChIP assay revealed binding of p65 to miR-21 promoter. NFκB activation was also required for the nuclear translocation of Smad 4 protein and subsequent direct interaction of Smad proteins with primary miR-21 as revealed by RNA-IP studies. Our results show that manipulation of TGFβ1-NFκB-miR-21 pathway could serve as an innovative approach towards therapeutics to heal diabetic ulcers.
authors.affiliation: Department of Applied Physiology, School of Medicine, University of Miyazaki, Miyazaki, Japan. Electronic address: radharao@med.miyazaki-u.ac.jp.
authors.forename: Radha
authors.initials: R
authors.lastname: Madhyastha
authors.affiliation: Department of Applied Physiology, School of Medicine, University of Miyazaki, Miyazaki, Japan.
authors.forename: HarishKumar
authors.initials: H
authors.lastname: Madhyastha
authors.affiliation: Department of Applied Physiology, School of Medicine, University of Miyazaki, Miyazaki, Japan.
authors.forename: Yutthana
authors.initials: Y
authors.lastname: Pengjam
authors.affiliation: Department of Applied Physiology, School of Medicine, University of Miyazaki, Miyazaki, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Nakajima
authors.affiliation: Department of Applied Physiology, School of Medicine, University of Miyazaki, Miyazaki, Japan.
authors.forename: Sayuri
authors.initials: S
authors.lastname: Omura
authors.affiliation: Department of Applied Physiology, School of Medicine, University of Miyazaki, Miyazaki, Japan.
authors.forename: Masugi
authors.initials: M
authors.lastname: Maruyama
mesh_terms: D000818:Animals; D017719:Diabetic Foot / Q000628:therapy; D005947:Glucose / Q000008:administration & dosage*; D020672:I-kappa B Proteins / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis*; D016328:NF-kappa B / Q000378:metabolism*; D041681:NIH 3T3 Cells; D021122:Protein Subunits / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D014945:Wound Healing / Q000187:drug effects
chemical_list: D020672:I-kappa B Proteins; C531950:MIRN21 microRNA, mouse; D035683:MicroRNAs; D016328:NF-kappa B; D021122:Protein Subunits; D053773:Transforming Growth Factor beta1; D005947:Glucose
keywords: Diabetic wound healing; MicroRNA; NFκB; TGFβ1; miR-21