title: Effects of eplerenone on the activation of matrix metalloproteinase-2 stimulated by high glucose and interleukin-1β in human cardiac fibroblasts.
abstract: The aim of this study was to determine the influence of high glucose (HG) and interleukin (IL)-1β on human cardiac fibroblast (HCF) functions, and to evaluate the effects of eplerenone in these responses. HCFs were cultured in normal or HG media in the absence or presence of IL-1β and/or eplerenone. We assessed matrix metalloproteinase-2 (MMP-2) activity in the supernatant by in-gel zymography, and determined mRNA expression levels of MMP-2 and tissue inhibitor of metalloproteinase-2 (TIMP-2) by reverse transcription-polymerase chain reaction. Equimolar D-mannitol was used as an osmotic control. HG stimulated MMP-2 activity and promoted MMP-2 mRNA synthesis. Increased effects were also observed in equimolar D-mannitol treatments, but these effects were weaker compared to those of glucose. The combination of HG and IL-1β resulted in a 2-fold increase in MMP-2 activity and mRNA expression compared with HG or IL-1β alone. Increases in HG- or IL-1β-induced MMP-2 activity and mRNA expression were blocked by eplerenone. Neither HG nor IL-1β affected TIMP-2 mRNA expression. HG increased MMP-2 activity by regulation of MMP- 2 mRNA expression in HCFs through osmotic and non-osmotic pathways. Synergistic effects of IL-1β added to HG media on MMP-2 activity and mRNA expression were observed in HCFs. Eplerenone normalized the effect of MMP-2 activity and HG- or IL-1β-induced expression in HCFs.
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Faculty of Medical Sciences, University of Fukui, Shimoaizuki, Matsuoka, Eiheiji-Cho, Fukui, Japan.
authors.forename: J F
authors.initials: JF
authors.lastname: Chi
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Faculty of Medical Sciences, University of Fukui, Shimoaizuki, Matsuoka, Eiheiji-Cho, Fukui, Japan hiroyasu@126.com.
authors.forename: H
authors.initials: H
authors.lastname: Uzui
authors.affiliation: Department of Cardiology, Shaoxing People's Hospital, Shaoxing Hospital of Zhejiang University, Shaoxing, Zhejiang, China.
authors.forename: H Y
authors.initials: HY
authors.lastname: Guo
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Faculty of Medical Sciences, University of Fukui, Shimoaizuki, Matsuoka, Eiheiji-Cho, Fukui, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Ueda
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, Faculty of Medical Sciences, University of Fukui, Shimoaizuki, Matsuoka, Eiheiji-Cho, Fukui, Japan.
authors.forename: J D
authors.initials: JD
authors.lastname: Lee
mesh_terms: D002460:Cell Line; D003470:Culture Media; D004789:Enzyme Activation / Q000187:drug effects; D000077545:Eplerenone; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D018093:Gelatinases / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology*; D008353:Mannitol / Q000494:pharmacology*; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D009206:Myocardium / Q000166:cytology / Q000201:enzymology; D009994:Osmolar Concentration; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013148:Spironolactone / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D003470:Culture Media; D053583:Interleukin-1beta; D012333:RNA, Messenger; D013148:Spironolactone; D008353:Mannitol; D000077545:Eplerenone; D018093:Gelatinases; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D005947:Glucose