Cellular Physiology and Biochemistry

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb

52

Ursu et al.:December Heme Oxygenase-1 Accepted: 12, 2013 in Enteroviral Myocarditis 1421-9778/14/0331-0052$39.50/0 This is an Open Access article licensed under the terms of the Creative Commons AttributionNonCommercial 3.0 Unported license (CC BY-NC) (www.karger.com/OA-license), applicable to the online version of the article only. Distribution permitted for non-commercial purposes only.

Original Paper

Heme Oxygenase-1 Mediates Oxidative Stress and Apoptosis in Coxsackievirus B3-Induced Myocarditis Oana N. Ursu

Martina Sauter Nicole Ettischer

Reinhard Kandolf

Karin Klingel

Institute for Pathology and Neuropathology, Department of Molecular Pathology, University Hospital Tübingen, Tübingen, Germany

Key Words 0\RFDUGLWLV‡+HPHR[\JHQDVH‡)UHHUDGLFDOV‡$SRSWRVLV‡,URQ Abstract Background: +HPH R[\JHQDVH +2  ZKLFK LV VXJJHVWHG WR SOD\ D UROH LQ GHIHQGLQJ WKH RUJDQLVP DJDLQVW R[LGDWLYH VWUHVVPHGLDWHG LQMXULHV FDQ EH LQGXFHG E\ GLYHUVH IDFWRUV LQFOXGLQJYLUXVHVDQGLURQ$VFR[VDFNLHYLUXV%&9% LQIHFWHG6:5-PLFHVXVFHSWLEOHIRU FKURQLFP\RFDUGLWLVZHUHIRXQGWRKDYHDVLJQLÀFDQWLURQLQFRUSRUDWLRQDQG+2XSUHJXODWLRQ LQWKHP\RFDUGLXPZHDLPHGWRLQYHVWLJDWHWKHPROHFXODULQWHUSOD\EHWZHHQ+2H[SUHVVLRQ and iron homeostasis in the outcome of viral myocarditis. Methods and Results: In susceptible 6:5-PLFHEXWQRWLQUHVLVWDQW&%/PLFHZHREVHUYHGDWODWHUVWDJHVRI&9% myocarditis VLJQLÀFDQW LURQ GHSRVLWV LQ PDFURSKDJHV DQG DOVR LQ FDUGLRP\RF\WHV ZKLFK ZHUH VSDWLDOO\ DVVRFLDWHGZLWKR[LGDWLYHVWUHVVXSUHJXODWLRQRI+2DQGFDVSDVHDFWLYDWLRQ+2ZKLFKLV DOVRH[SUHVVHGLQFXOWLYDWHG5$:PDFURSKDJHVXSRQLQFXEDWLRQZLWKLURQDQGRU&9% FRXOGEHGRZQUHJXODWHGE\LQKLELWLRQRI12L126XVLQJ/1$0(0RUHRYHUVSHFLÀFLQKLELWLRQ RI+2E\WLQPHVRSRUSK\ULQUHYHDOHGDVXSSUHVVLRQRIVXSHUR[LGHSURGXFWLRQLQLURQDQG RU&9%WUHDWHGPDFURSKDJHV7KHPROHFXODUUHODWLRQVKLSRI+2DQGFDVSDVHDFWLYDWLRQ ZDVSURYHQE\GRZQUHJXODWLRQZLWK+2VL51$LQLURQDQGRU&9%WUHDWHGFXOWLYDWHGFHOOV Importantly, iron was found to increase viral replication in vitro. Conclusion: These results LQGLFDWHWKDW+2LQGXFHVDSDUDFULQHVLJQDOOLQJLQPDFURSKDJHVYLDUHDFWLYHR[\JHQVSHFLHV production, mediating apoptosis of heart muscle cells at later stages of myocarditis. Notably, LQJHQHWLFDOO\VXVFHSWLEOHPLFHLURQSRWHQWLDWHVWKHGHWULPHQWDOHIIHFWVRI&9%E\WKH12 HO-1 pathway, thus increasing cardiac pathogenicity.

Prof. Dr. Karin Klingel

Institute for Pathology and Neuropathology, Department of Molecular Pathology, /LHEHUPHLVWHUVWU7ELQJHQ*HUPDQ\ 7HO)D[(0DLONDULQNOLQJHO#PHGXQLWXHELQJHQGH

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Copyright © 2014 S. Karger AG, Basel

Cellular Physiology and Biochemistry

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb

53

Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

Introduction

Coxsackieviruses of group B are members of the Picornaviridae family which have „‡‡‹†‡–‹ϐ‹‡†ƒ•ƒŒ‘”…ƒ—•‡‘ˆƒ…—–‡ƒ†…Š”‘‹…ˆ‘”•‘ˆ›‘…ƒ”†‹–‹•ȏͳȐǤ …ƒ•‡‘ˆƒ ineffective host immune response, these enteroviruses may persist beyond the acute phase ‘ˆ–Š‡‹ˆ‡…–‹‘ƒ†‹†—…‡ƒ‘‰‘‹‰‹ϐŽƒƒ–‹‘™Š‹…Šƒ›Ž‡ƒ†–‘†‹Žƒ–‡†…ƒ”†‹‘›‘’ƒ–Š› ƒ› ›‡ƒ”• ƒˆ–‡” –Š‡ ‹‹–‹ƒŽ ‹ˆ‡…–‹‘ ȏʹȐǤ ‘š•ƒ…‹‡˜‹”—•‡•ǡ ‹…Ž—†‹‰ ͵ǡ ƒˆˆ‡…– –Š‡ cardiomyocytes directly as a consequence of viral replication, but also elicit immune cell ‹ϐ‹Ž–”ƒ–‡• †‡–‡”‹‹‰ –Š‡ †‹ˆˆ‡”‡– ‘—–…‘‡ ‘ˆ –Š‡ ‹ϐŽƒƒ–‘”› Š‡ƒ”– †‹•‡ƒ•‡ ƒ– Žƒ–‡” •–ƒ‰‡• ‘ˆ –Š‡ †‹•‡ƒ•‡ ȏ͵Ȑ Ǥ ‡ ‹’‘”–ƒ– …‡ŽŽ –›’‡ ™Š‹…Š ‹• †‡…‹•‹˜‡ ‹ –Š‡ ‡ƒ”Ž› ƒ–‹˜‹”ƒŽ innate immune response are macrophages which wereˆ‘—†–‘†‡ϐ‹‡–Š‡•—•…‡’–‹„‹Ž‹–›–‘ ͵›‘…ƒ”†‹–‹•˜‹ƒ†‹ˆˆ‡”‡–‹ƒŽ…›–‘‹‡‡š’”‡••‹‘ȏͶȐǤthe other hand, macrophages ‡š’”‡••‹†—…‹„Ž‡‹–”‹…‘š‹†‡•›–Šƒ•‡ȋ‹Ȍǡ™Š‹…Š™ƒ•ˆ‘—†–‘…‘–”‹„—–‡•‹‰‹ϐ‹…ƒ–Ž› –‘‘‰‘‹‰†ƒƒ‰‡„›‹–”‹…‘š‹†‡ȋȌ’”‘†—…–‹‘ƒ–Žƒ–‡”•–ƒ‰‡•‘ˆ͵‹ˆ‡…–‹‘ȏͷȐǤ ‹•ƒ™‡ŽŽ‘™‡†‹ƒ–‘”ˆ‘”–Š‡‰‡‡”ƒ–‹‘‘ˆ”‡ƒ…–‹˜‡‘š›‰‡•’‡…‹‡•ȋȌƒ†”‡ƒ…–‹˜‡ ‹–”‘‰‡•’‡…‹‡•ȋȌǡ™Š‹…Šƒ”‡”‡•’‘•‹„Ž‡ˆ‘”–Š‡‰‡‡”ƒ–‹‘‘ˆ‘š‹†ƒ–‹˜‡•–”‡••‹–Š‡ ‹ϐŽƒ‡† ›‘…ƒ”†‹— ȏ͸ȐǤ Ž•‘ǡ ͵ ”‡’Ž‹…ƒ–‹‘ ‹ cardiomyocytes is associated with –Š‡ ’”‘†—…–‹‘ ‘ˆ  ‹ –Š‡•‡ …‡ŽŽ• ȏ͹ȐǤ –”ƒ…‡ŽŽ—Žƒ”  Ž‡˜‡Ž• ƒ”‡ ‰‡‡”ƒŽŽ› …‘–”‘ŽŽ‡† „›†‹ˆˆ‡”‡–ƒ–‹‘š‹†ƒ–†‡ˆ‡•‡‡…Šƒ‹••ǡŠ‘™‡˜‡”ǡ‘˜‡”’”‘†—…–‹‘ƒ›”‡•—Ž–‹ †‡–”‹‡–ƒŽ ‹–”ƒ…‡ŽŽ—Žƒ” †‹•–—”„ƒ…‡•Ǥ ‡ ‹’‘”–ƒ– ‘Ž‡…—Ž‡ ‹ –he cellular response to stress/oxidant stimuli is Š‡‡‘š›‰‡ƒ•‡ǦͳȋǦͳȌǡ–Š‡‹†—…‹„Ž‡‹•‘ˆ‘”‘ˆǡ™Š‹…Š catalyzes the rate-limiting step of heme oxidation to biliverdin, carbon monoxide, and ˆ”‡‡ ˆ‡””‘—• ‹”‘ ȏͺȐǤ Ǧͳǡ –Š‡ Š‡ƒ– •Š‘…Ȁ•–”‡•• …‘‰ƒ–‡ ‘ˆ –Š‡ Š‡ƒ– •Š‘… ’”‘–‡‹ ͵ʹ family of proteins, is expressed in response to a variety of stimuli that are associated with ‘š‹†ƒ–‹˜‡ •–”‡•• ƒ† ‹ϐŽƒƒ–‹‘ ȏͻȐǤ Ž•‘ǡ Ǧͳ  ƒ† ’”‘–‡‹ ‡š’”‡••‹‘ ™‡”‡ ˆ‘—† –‘ „‡ ‹…”‡ƒ•‡† „› ‹–”ƒ…‡ŽŽ—Žƒ” ‹”‘ ‘˜‡”Ž‘ƒ† ȏͳͲȐǤ ”‘ ‹• †‡•…”‹„‡† –‘ ‹’”‘˜‡ …ƒ”†‹ƒ…ˆ—…–‹‘‹’ƒ–‹‡–•™‹–Š…Š”‘‹…Š‡ƒ”–ˆƒ‹Ž—”‡ȋ Ȍ–Šƒ–ƒ”‡’”‘‡–‘†‡˜‡Ž‘’‹”‘ †‡ϐ‹…‹‡…› ȏͳͳȐǡ Š‘™‡˜‡” ‹– ƒ› ƒŽ•‘ „‡ …ƒ”†‹‘–‘š‹… —†‡” ‘˜‡”Ž‘ƒ† …‘†‹–‹‘• ȏͳʹȐǤ ”‘Ǧ overload cardiomyopathy is an important cause of heart failure and involves susceptibility –‘ƒ””Š›–Š‹ƒ•ƒ†ƒŽƒ–‡Ǧ•–ƒ‰‡†‹Žƒ–‡†…ƒ”†‹‘›‘’ƒ–Š›Ǥ–Žƒ–‡”•–ƒ‰‡•‘ˆ–Š‹•†‹•‡ƒ•‡ǡ–Š‡ disturbed iron metabolism in myocytes induces remodelling processes of the heart, features ™Š‹…Šƒ”‡ƒŽ•‘‘„•‡”˜‡†ƒ•ƒ…‘•‡“—‡…‡‘ˆ‡–‡”‘˜‹”—•Ǧ‹†—…‡†…Š”‘‹…›‘…ƒ”†‹–‹•ȏͳ͵ȐǤ –‡”‡•–‹‰Ž›ǡ„‘–Š͵ƒ†‹”‘ƒ”‡‘™–‘‡Šƒ…‡’”‘†—…–‹‘Ǥ‘™‡˜‡”ǡ–Š‡”‡ƒ”‡ no data available investigating the molecular interplay between enteroviral infection and ‹”‘‡–ƒ„‘Ž‹•‹–Š‡Š‡ƒ”–ǤŠ—•ǡ–Š‡ƒ‹‘ˆ–Š‹••–—†›™ƒ•–‘•–—†›–Š‡‹–‡”ƒ…–‹‘•‘ˆ ‡–‡”‘˜‹”—•”‡’Ž‹…ƒ–‹‘ǡ’”‘†—…–‹‘ǡ‹”‘‘˜‡”Ž‘ƒ†ƒ†Ǧͳ‡š’”‡••‹‘‹”‡Žƒ–‹‘–‘ –Š‡‘—–…‘‡‘ˆ›‘…ƒ”†‹–‹•‹͵Ǧ‹ˆ‡…–‡†‹…‡Ǥ Materials and Methods

Virus ͵—•‡†‹–Š‹••–—†›™ƒ•†‡”‹˜‡†ˆ”‘–Š‡‹ˆ‡…–‹‘—•……‘’›‘ˆ–Š‡…ƒ”†‹‘–”‘’‹…ƒ…›•–”ƒ‹ ƒ†˜‹”—••–‘…•ƒ†’Žƒ“—‡ƒ••ƒ›•™‡”‡’‡”ˆ‘”‡†ƒ•’”‡˜‹‘—•Ž›†‡•…”‹„‡†ȏͳͶȐǤ

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Mice and infection Ȁ ƒ†ͷ͹Ȁ͸‹…‡ǡ‘”‹‰‹ƒŽŽ›’—”…Šƒ•‡†ˆ”‘ ƒ…•‘ƒ„‘”ƒ–‘”‹‡•ȋƒ”ƒ”„‘”ǡȌǡ™‡”‡‡’– —†‡”•’‡…‹ϐ‹…’ƒ–Š‘‰‡Ǧˆ”‡‡…‘†‹–‹‘•ƒ––Š‡ƒ‹ƒŽˆƒ…‹Ž‹–‹‡•‘ˆ–Š‡‡’ƒ”–‡–‘ˆ‘Ž‡…—Žƒ”ƒ–Š‘Ž‘‰›ǡ ‹˜‡”•‹–› ‘•’‹–ƒŽ ò„‹‰‡ǡ ƒ† ‡š’‡”‹‡–• ™‡”‡ …‘†—…–‡† ƒ……‘”†‹‰ –‘ –Š‡ ‡”ƒ Žƒ™ ”‡‰ƒ”†‹‰ ƒ‹ƒŽ’”‘–‡…–‹‘ȋ’‡”‹–‘ǤͳȀͲͺȌǤ––Š‡ƒ‰‡‘ˆͶ–‘ͷ™‡‡•ǡ‹…‡™‡”‡‹–”ƒ’‡”‹–‘‡ƒŽŽ›‹‘…—Žƒ–‡† ™‹–ŠͳͲͷ’Žƒ“—‡Ǧˆ‘”‹‰—‹–•‘ˆ’—”‹ϐ‹‡†͵ƒ•’”‡˜‹‘—•Ž›†‡•…”‹„‡†ȏͳͶȐǤ‹ƒŽ•‘ˆ‡ƒ…Š•–”ƒ‹ȋηͷȌ ™‡”‡•ƒ…”‹ϐ‹…‡†ƒ–†ƒ›•Ͷǡͺǡͳʹƒ†ʹͺ’‘•–‹ˆ‡…–‹‘ȋ’Ǥ‹ǤȌƒ†Š‡ƒ”–•ǡ•’Ž‡‡ƒ†•‡”—™‡”‡…‘ŽŽ‡…–‡†ˆ‘” ƒƒŽ›•‹•Ǥ ‘”Š‹•–‘Ž‘‰›ǡ’ƒ”–•‘ˆ–Š‡…‘ŽŽ‡…–‡†‘”‰ƒ•™‡”‡ϐ‹š‡†‹ͶΨ’ƒ”ƒˆ‘”ƒŽ†‡Š›†‡ƒ†‡„‡††‡†‹ ’ƒ”ƒˆϐ‹Ǥ ‘”“—ƒ–‹–ƒ–‹˜‡ƒƒŽ›•‡•ǡ’ƒ”–•‘ˆ–Š‡‘”‰ƒ•™‡”‡•ƒ’Ǧˆ”‘œ‡ƒ†•–‘”‡†ƒ–ǦͺͲιǤ

Cellular Physiology and Biochemistry

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb

54

Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

Histology and immunohistochemical staining ƒ”ƒˆϐ‹Ǧ‡„‡††‡†Š‡ƒ”–•™‡”‡…—–‹ͷǦɊm-thick tissue sections and stained with hematoxylin and ‡‘•‹–‘ƒ••‡••›‘…ƒ”†‹ƒŽ‹Œ—”›ƒ†‹ϐŽƒƒ–‹‘Ǥ ‘”‡˜ƒŽ—ƒ–‹‘‘ˆϐ‹„”‘•‹•ǡƒ••‘–”‹…Š”‘‡•–ƒ‹‹‰ ™ƒ• ƒ’’Ž‹‡† ™Š‹Ž‡ ‹”‘ †‡’‘•‹–• ™‡”‡ †‡–‡…–‡† „› ”—••‹ƒ „Ž—‡ •–ƒ‹‹‰Ǥ ‘” ‹—‘Š‹•–‘…Š‡‹•–”›ǡ –‹••—‡•‡…–‹‘•™‡”‡‹…—„ƒ–‡†ˆ‘”ͳŠ‘—”ƒ–ʹͷι™‹–Š”ƒ„„‹–ƒ–‹Ǧ‘—•‡ƒ–‹„‘†‹‡•”‡…‘‰‹œ‹‰—”‹‡ Ǧͳ ȋͳǣͳͲͲǡ „…ƒǡ Ȍǡ …Ž‡ƒ˜‡† …ƒ•’ƒ•‡Ǧ͵ ȋͳǣͶͲͲǡ ‡ŽŽ ‹‰ƒŽ‹‰ ‡…Š‘Ž‘‰›ǡ ƒ˜‡”•ǡ ǡ ȌǤ ‘–”‘Ž•—•‹‰‘”ƒŽ”ƒ„„‹–•‡”—™‡”‡”—–‘‡š…Ž—†‡‘•’‡…‹ϐ‹…•–ƒ‹‹‰ǤŽ‹†‡•™‡”‡’”‘…‡••‡†—•‹‰ •–”‡’–‘’Ž‡šȋƒ‘ǡƒ„—”‰ǡ ‡”ƒ›Ȍƒ†͵ǡ͵Ʈ†‹ƒ‹‘„‡œ‹†‹‡ȋ‹‘œ‘Žǡ…Š‹‰ǡ ‡”ƒ›Ȍƒ• •—„•–”ƒ–‡Ǥ‹•—ƒŽ‹œƒ–‹‘‘ˆ͵ͳ’”‘–‡‹™ƒ•†‘‡„›‹…—„ƒ–‹‰†‡’ƒ”ƒˆϐ‹‹œ‡†–‹••—‡•‡…–‹‘•™‹–Šƒ ‘‘…Ž‘ƒŽƒ–‹„‘†›•’‡…‹ϐ‹…ˆ‘”͵ͳȋͳǣͶͲͲǡ‡†‹ƒ‰‘•–ǡ‡—–Ž‹‰‡ǡ ‡”ƒ›Ȍˆ‘”ͳŠ‘—”ƒ–”‘‘ –‡’‡”ƒ–—”‡Ǥ—„•‡“—‡–Ž›ǡ–Š‡•‡…–‹‘•™‡”‡’”‘…‡••‡†™‹–Š–Š‡ƒš‘‘‘—•‡‘‘—•‡‘Ž›‡” ‡–‡…–‹‘ ‹– ȋƒš‹•‹‘ǡ ƒ•Š‹‰–‘ǡ Ȍ ˆ‘ŽŽ‘™‡† „›  •—„•–”ƒ–‡ ȋ‹ƒ”‹•ǡ‘••‡Š‡‹ǡ ‡”ƒ›ȌǤ Š‡–‹••—‡•‡…–‹‘•™‡”‡…‘—–‡”•–ƒ‹‡†™‹–ŠŠ‡ƒ–‘š›Ž‹Ǥ Quantitative real- time RT-PCR —”‹‡Š‡ƒ”––‹••—‡•™‡”‡Ž›•‡†‹”‹ˆƒ•–ȋ‡“Žƒ„ǡ”Žƒ‰‡ǡ ‡”ƒ›Ȍƒ……‘”†‹‰–‘–Š‡ƒ—ˆƒ…–—”‡”ǯ• ‹•–”—…–‹‘•ǡƒ†–Š‡‹–‡‰”‹–›ƒ†…‘…‡–”ƒ–‹‘‘ˆ–Š‡–‘–ƒŽ™ƒ•ƒ••‡••‡†„›ƒ‰ƒ”‘•‡‰‡Ž‡Ž‡…–”‘’Š‘”‡•‹• ƒ†„›’Š‘–‘•’‡…–”‘‡–”›ǡ”‡•’‡…–‹˜‡Ž›ǤȋʹͲͲ‰Ȍ™ƒ•—•‡†–‘’‡”ˆ‘”‘‡•–‡’“—ƒ–‹–ƒ–‹˜‡”‡ƒŽǦ–‹‡ ”‡˜‡”•‡Ǧ–”ƒ•…”‹’–‹‘ȋƒ“ƒ‡Ǧ–‡’Ǧƒ•–‡”‹š‡ƒ‰‡–•‹–ǡ’’Ž‹‡†‹‘•›•–‡•ǡ ‘•–‡” ‹–›ǡȌƒ––Š‡ƒ’’”‘’”‹ƒ–‡ƒ‡ƒŽ‹‰–‡’‡”ƒ–—”‡ˆ‘”ͶͲ…›…Ž‡•Ǥƒ–ƒƒƒŽ›•‹•™ƒ•’‡”ˆ‘”‡†ƒ•”‡Žƒ–‹˜‡ “—ƒ–‹ϐ‹…ƒ–‹‘‹”‡Žƒ–‹‘–‘–Š‡‡š’”‡••‹‘‘ˆ–Š‡Š‘—•‡‡‡’‹‰‰‡‡Š›’‘šƒ–Š‹Ǧ‰—ƒ‹Ǧ’Š‘•’Š‘”‹„‘•›ŽǦ –”ƒ•ˆ‡”ƒ•‡ȋȌƒ•‹–‡”ƒŽ•–ƒ†ƒ”†ǤŠ‡ƒ„•‘Ž—–‡—„‡”‘ˆ͵˜‹”ƒŽ‰‡‘‡•™ƒ•†‡–‡”‹‡†‹ ”‡Žƒ–‹‘–‘ƒ‡š–‡”ƒŽ˜‹”—••–ƒ†ƒ”†—•‹‰ƒ–‡”‘Ǧ’”‹‡”•ƒ†’”‘„‡•Ǥ’‡…‹ϐ‹…’”‹‡”•ƒ†’”‘„‡• ™‡”‡’—”…Šƒ•‡†ˆ”‘ ‹‘–‡…Šȋ„‡”•„‡”‰ǡ ‡”ƒ›ȌǤ In situ hybridization (ISH) ͵ ’‘•‹–‹˜‡Ǧ•–”ƒ† ‰‡‘‹…  ‹ –‹••—‡• ™ƒ• †‡–‡…–‡† —•‹‰ •‹‰Ž‡Ǧ•–”ƒ†‡† ͵ͷǦŽƒ„‡Ž‡†  ’”‘„‡•ƒ•’”‡˜‹‘—•Ž›†‡•…”‹„‡†ȏͳͶȐǤ ‘”†‡–‡…–‹‘‘ˆ‹‹–‹••—‡•ǡ͵ͷǦŽƒ„‡Ž‡†ƒ–‹Ǧ•‡•‡ƒ†•‡•‡ ‹’”‘„‡•™‡”‡•›–Š‡•‹œ‡†ƒ•’”‡˜‹‘—•Ž›†‡•…”‹„‡†ȏͷȐǤ ‘”†‡–‡…–‹‘‘ˆǦͳǡƒ–‹Ǧ•‡•‡ƒ† •‡•‡’”‘„‡•™‡”‡•›–Š‡•‹œ‡†„ƒ•‡†‘ƒǦͳ’Žƒ•‹†ȋ‹ƒ ‡‡•ǡ‡”Ž‹ǡ ‡”ƒ›ȌǤ”‡–”‡ƒ–‡–ǡ Š›„”‹†‹œƒ–‹‘ǡ ƒ† ™ƒ•Š‹‰ ’”‘…‡†—”‡• ‘ˆ †‡™ƒš‡† ͷɊ ’ƒ”ƒˆϐ‹ –‹••—‡ •‡…–‹‘• ™‡”‡ ’‡”ˆ‘”‡† ƒ• ’”‡˜‹‘—•Ž›†‡•…”‹„‡†ȏͳͶȐǤŽ‹†‡’”‡’ƒ”ƒ–‹‘•™‡”‡•—„Œ‡…–‡†–‘ƒ—–‘”ƒ†‹‘‰”ƒ’Š›ǡ‡š’‘•‡†ˆ‘”͵™‡‡•ƒ– Ͷιǡƒ†…‘—–‡”•–ƒ‹‡†™‹–ŠŠƒ‡ƒ–‘š›Ž‹Ȁ‡‘•‹Ǥ

Cell culture ‘”‡•–‡”„Ž‘–ƒ†‹—‘ϐŽ—‘”‡•…‡…‡‡š’‡”‹‡–•ǡ‡ƒ…‡ŽŽ•ƒ†–Š‡ƒ…”‘’Šƒ‰‡…‡ŽŽŽ‹‡ ʹ͸ͶǤ͹ȋ –ƒ†ƒ”†•ǡȌ™‡”‡—•‡†Ǥ‘–Š…‡ŽŽŽ‹‡•™‡”‡•‡”—Ǧ•–ƒ”˜‡†‘˜‡”‹‰Š–ƒ†…‡ŽŽ•™‡”‡ ‹–‡”ˆ‡”‘ȋ ȌɀǦ•–‹—Žƒ–‡†ȋͳͲͲȀŽǡƬ›•–‡•ǡ ‡”ƒ›Ȍˆ‘”ͳŠ‘—”–Š‡‡š–†ƒ›Ǥ ‘”…‘Ǧ…—Ž–—”‡ ƒ† ‹”‘ ‡š’‡”‹‡–•ǡ ͳ‘ŽȀ ‘ˆ •‘Ž—„Ž‡ ‹”‘ ȋˆ‡””‹… ƒ‘‹— …‹–”ƒ–‡Ȍ ™ƒ• ƒ††‡† –‘ –Š‡  ʹ͸ͶǤ͹ …‡ŽŽ•ȋ–‘ƒϐ‹ƒŽ…‘…‡–”ƒ–‹‘‘ˆͳͲͲρ‘ŽȀȌƒ†ƒŽŽ‘™‡†–‘’‡”‡ƒ–‡–Š‡‡„”ƒ‡ˆ‘”ͳŠ‘—”ƒ–͵͹ιǤ ‹—Ž–ƒ‡‘—•Ž›ǡ‡ƒ…‡ŽŽ•™‡”‡͵Ǧ‹ˆ‡…–‡†ȋ ‘ˆͷȌˆ‘”ͳŠ‘—”„›ƒ††‹‰–Š‡˜‹”—•–‘–Š‡‡†‹— ™‹–Š‘—– ˆ—”–Š‡” ™ƒ•Š‡•Ǥ ˆ–‡” ͳ Š‘—”ǡ ‡ƒ …‡ŽŽ• ™‡”‡ ƒ††‡† –‘ –Š‡  ʹ͸ͶǤ͹ …‡ŽŽ• ƒ† ‹˜‡•–‹‰ƒ–‡† ƒ– •’‡…‹ϐ‹…–‹‡’‡”‹‘†•Ǥ•…‘–”‘Ž•ǡ‹”‘Ǧˆ”‡‡ƒ†—‹ˆ‡…–‡†…‡ŽŽ•™‡”‡—•‡†Ǥ ‘”‹‹Š‹„‹–‹‘‡š’‡”‹‡–•ǡ

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Dihydroethidium staining (DHE)  •‹–— ’”‘†—…–‹‘ ‘ˆ •—’‡”‘š‹†‡ ƒ‹‘ ™ƒ• ƒ••‡••‡† ‹ ͷρ ˆ”‘œ‡ Š‡ƒ”– –‹••—‡ •‡…–‹‘• ™‹–Š –Š‡ ‘š‹†ƒ–‹˜‡ϐŽ—‘”‡•…‡–†›‡†‹Š›†”‘‡–Š‹†‹—ȋǡ ˜‹–”‘‰‡ǡƒ”Ž•”—Š‡ǡ ‡”ƒ›ȌǤ‡…–‹‘•™‡”‡ϐ‹š‡†ˆ‘” ͳͲ‹—–‡•‹…‘Ž†ƒ…‡–‘‡ǡ™ƒ•Š‡†‹ƒ†‹…—„ƒ–‡†‹ȋϐ‹ƒŽ…‘…‡–”ƒ–‹‘‘ˆͷρ‘ŽȀȌˆ‘”ͳͷ ‹—–‡•ƒ–͵͹郐†…‘—–‡”•–ƒ‹‡†™‹–Š Ǥ Ž—‘”‡•…‡…‡‹ƒ‰‡•™‡”‡‘„–ƒ‹‡†™‹–Šƒ‡‹••š‹‘˜‡”– ʹͲͲͲ ‹…”‘•…‘’‡Ǥ ‘” •—’‡”‘š‹†‡ †‡–‡…–‹‘ ‹ …‡ŽŽ•ǡ Ž‹˜‡Ǧ‹ƒ‰‹‰ ‡š’‡”‹‡–• ™‡”‡ ’‡”ˆ‘”‡† ‹ ‹”‘Ǧ –”‡ƒ–‡†ƒ†͵Ǧ‹ˆ‡…–‡†…‘…—Ž–—”‡‘ˆ‡ƒ…‡ŽŽ•ƒ†ʹ͸ͶǤ͹ƒ…”‘’Šƒ‰‡•Ǥ ‘”†‡”–‘†‹ˆˆ‡”‡–‹ƒ–‡ „‡–™‡‡  ʹ͸ͶǤ͹ ƒ…”‘’Šƒ‰‡• ƒ† ‡ƒ …‡ŽŽ•ǡ ƒ…”‘’Šƒ‰‡• ™‡”‡ Žƒ„‡Ž‡† ™‹–Š ‡ŽŽ”ƒ…‡” ”‡‡ ȋ‘œƒǡ ‡”ƒ›Ȍ„‡ˆ‘”‡Žƒ„‡Ž‹‰Ǥ ‘”–Š‡Ž‹˜‡Ǧ‹ƒ‰‹‰ǡ…‡ŽŽ•™‡”‡‡’–‹ƒ‹…—„ƒ–‹‘…Šƒ„‡”‘ –Š‡‹…”‘•…‘’‡ƒ–͵͹ιǡͷΨʹƒ†‹Š—‹†ƒ–‘•’Š‡”‡Ǥ

Cellular Physiology and Biochemistry

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb

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Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

 ʹ͸ͶǤ͹ …‡ŽŽ• ™‡”‡ ’”‡Ǧ–”‡ƒ–‡† ˆ‘” ͳ Š‘—” ™‹–Š ͳ‘ŽȀ  Ǧ‹–”‘ǦǦƒ”‰‹‹‡ ‡–Š›Ž‡•–‡” ȋǦǡ ‹‰ƒǦŽ†”‹…Šǡ ‡”ƒ›ȌǤ ‘”Ǧͳ‹Š‹„‹–‹‘‡š’‡”‹‡–•ǡʹ͸ͶǤ͹ƒ…”‘’Šƒ‰‡•™‡”‡’”‡Ǧ–”‡ƒ–‡†™‹–Š ʹͲ‘ŽȀ –‹ ‡•‘’‘”’Š›”‹ ȋȌ ˆ‘” ͸ Š‘—”• ȋƒ›ƒ Š‡‹…ƒŽ•ǡ Ȍ „‡ˆ‘”‡ ƒ††‹‰ ‹”‘ ƒ†Ȁ‘” ͵Ǧ‹ˆ‡…–‡†‡ƒ…‡ŽŽ•‘˜‡”‹‰Š–Ǥ‡ŽŽŽ›•ƒ–‡•™‡”‡„Ž‘––‡†‘ ‡„”ƒ‡•‘”—•‡†–‘‡š–”ƒ…–Ǥ Immunoblotting analysis ƒ’Ž‡•‘ˆˆ”‘œ‡—”‹‡Š‡ƒ”–•‘”…‡ŽŽŽ›•ƒ–‡•™‡”‡‹…‡†ƒ†Š‘‘‰‡‹œ‡†‹‹…‡…‘Ž†Ž›•‹•„—ˆˆ‡” ȋ‡ŽŽ‹‰ƒŽŽ‹‰‡…Š‘Ž‘‰›ǡƒ˜‡”•ǡǡȌ•—’’Ž‡‡–‡†™‹–Š‘’Ž‡–‡Ǧˆ”‡‡”‘–‡ƒ•‡ Š‹„‹–‘” ‘…–ƒ‹Ž ȋ‘…Š‡ǡ ƒŠ‡‹ǡ ‡”ƒ›Ȍ ƒ– ”‡…‘‡†‡† …‘…‡–”ƒ–‹‘•Ǥ ͵Ͳρ‰ ‘ˆ –Š‡ ‡š–”ƒ…–‡† ’”‘–‡‹• ™‡”‡•‡’ƒ”ƒ–‡†‘ƒͳͲΨ’‘Ž›ƒ…”›Žƒ‹†‡‰‡Žƒ†–”ƒ•ˆ‡””‡†–‘ ‡„”ƒ‡•Ǥˆ–‡”„Ž‘…‹‰™‹–ŠͷΨ ‘Ǧˆƒ–†”›‹Ž‹ˆ‘”ͳŠ‘—”ƒ–”‘‘–‡’‡”ƒ–—”‡ǡ„Ž‘–•™‡”‡‹…—„ƒ–‡†‘˜‡”‹‰Š–ƒ–Ͷι™‹–Š”ƒ„„‹– ƒ–‹Ǧ‹ȋ„…ƒǡȌǡ”ƒ„„‹–ƒ–‹Ǧͳȋ„…ƒǡȌ‘””ƒ„„‹–ƒ–‹ǦȾǦ–—„—Ž‹ȋ‡ŽŽ‹‰ƒŽŽ‹‰‡…Š‘Ž‘‰›ǡ ƒ˜‡”•ǡǡȌǤŽŽ’”‹ƒ”›ƒ–‹„‘†‹‡•™‡”‡—•‡†ƒ–ƒͳǣͳͲͲͲ†‹Ž—–‹‘Ǥ‡…‘†ƒ”›’‡”‘š‹†ƒ•‡Ǧ…‘Œ—‰ƒ–‡† ‰‘ƒ–ƒ–‹Ǧ”ƒ„„‹– ‰ ƒ–‹„‘†‹‡•ȋƒ–ƒ”—œ‹‘–‡…Š‘Ž‘‰›ǡƒ–ƒ”—œǡǡǡͳǣʹͲͲͲ‹Ȁͷؐ‘Ǧˆƒ– †”›‹ŽȌ™‡”‡—•‡†ˆ‘”…Š‡‹Ž—‹‡•…‡–’”‘–‡‹†‡–‡…–‹‘™‹–Š—’‡”‹‰ƒŽ‡•–‹…‘•—„•–”ƒ–‡ȋ‹‡”…‡ǡ ȌǤƒ”‰‡–’”‘–‡‹Ž‡˜‡Ž•™‡”‡“—ƒ–‹ϐ‹‡†„› ƒ‰‡ ‹ƒ‰‡ƒƒŽ›•‹••‘ˆ–™ƒ”‡ȋƒ–‹‘ƒŽ •–‹–—–‡•‘ˆ‡ƒŽ–Š (NIHȌǡ‡–Š‡•†ƒǡȌƒ†‘”ƒŽ‹œ‡†–‘ȾǦ–—„—Ž‹Ǥ Transfection with HO-1 siRNA ‡ƒ…‡ŽŽ•ȋʹšͳͲͶȌ™‡”‡‰”‘™‹ͻ͸Ǧ™‡ŽŽ•’Žƒ–‡ȋ—…ǡ‘•‹Ž†‡ǡ‡ƒ”Ȍ‹•‡”—ƒ†ƒ–‹„‹‘–‹…Ǧ ˆ”‡‡‡†‹—ƒ†–”ƒ•ˆ‡…–‡†ƒ–͸ͲǦ͹ͲΨ…‘ϐŽ—‡…›Ǥˆ–‡”™ƒ•Š‹‰™‹–Š’–‹Ǧȋ ˜‹–”‘‰‡ǡƒ”Ž•”—Š‡ǡ

‡”ƒ›Ȍ …‡ŽŽ• ™‡”‡ ‘˜‡”Žƒ‹† ™‹–Š –Š‡ –”ƒ•ˆ‡…–‹‘ ‹š–—”‡ …‘–ƒ‹‹‰ ”‘‘ ‡…–‹Ǧ•‹ ”‡ƒ‰‡– ȋͷρŽǡ ”‘‘‡ŽŽǡ ‡‹†‡Ž„‡”‰ǡ ‡”ƒ›Ȍ ƒ† …‘‡”…‹ƒŽŽ› ƒ˜ƒ‹Žƒ„Ž‡ •‹ ƒ‰ƒ‹•– ŠǦͳ ȋͳρ‰ǡ ƒ–ƒ ”—œ ‹‘–‡…Š‘Ž‘‰›ǡ ƒ–ƒ ”—œǡ ǡ Ȍ ‹ ƒ ϐ‹ƒŽ ˜‘Ž—‡ ‘ˆ ʹͲͲρŽ ’–‹ǦǤ ‹š Š‘—”• „‡ˆ‘”‡ ƒ••ƒ›ǡ ‹”‘ ȋͳͲͲρ‘ŽȀȌƒ†Ȁ‘”͵ȋƒ– ‘ˆͷȌ™‡”‡ƒ††‡†–‘–Š‡…‡ŽŽ•Ǥ•‹…‘–ƒ‹‹‰ƒ•…”ƒ„Ž‡†•‡“—‡…‡ ™ƒ•—•‡†ƒ•…‘–”‘ŽǤǦͳ•‹–”ƒ•ˆ‡…–‹‘‡ˆϐ‹…‹‡…›‹–Š‡”‡•’‡…–‹˜‡•ƒ’Ž‡•™ƒ•†‡–‡”‹‡†„›“Ǧ Ǥ —‘ϔŽ—‘”‡•…‡…‡ȋ Ȍ ‘ϐŽ—‡–͵Ǧ‹ˆ‡…–‡†ƒ†Ȁ‘”‹”‘–”‡ƒ–‡†‡ƒ…‡ŽŽ•‘”ʹ͸ͶǤ͹ƒ…”‘’Šƒ‰‡•™‡”‡ϐ‹š‡†‹ͶΨ ’ƒ”ƒˆ‘”ƒŽ†‡Š›†‡•‘Ž—–‹‘‹ȋͳͷ‹—–‡•ǡ”‘‘–‡’‡”ƒ–—”‡Ȍƒ†’‡”‡ƒ„‹Ž‹œ‡†„›ͲǡͳΨ”‹–‘ǦͳͲͲ •‘Ž—–‹‘ ‹  ȋʹͲ ‹—–‡•ǡ ”‘‘ –‡’‡”ƒ–—”‡ȌǤ ‡ŽŽ• ™‡”‡ ‹…—„ƒ–‡† ™‹–Š •’‡…‹ϐ‹… ’”‹ƒ”› ƒ–‹„‘†‹‡• ȋ”ƒ„„‹–Ǧͳǡ”ƒ„„‹–‹ȋ„…ƒǡƒ„”‹†‰‡ǡȌǡ”ƒ„„‹–…ƒ•’ƒ•‡Ǧ͵ȋ‡ŽŽ‹‰ƒŽŽ‹‰‡…Š‘Ž‘‰›ǡƒ˜‡”•ǡ ǡȌƒ†‘š͵ͳʹȋ͵ͳȌȋ‡†‹ƒ‰‘•–ǡ‡—–Ž‹‰‡ǡ ‡”ƒ›Ȍǡ†‹Ž—–‡†ͳǣͳͲͲˆ‘”Ǧͳǡ‹ ƒ†ͳǣͶͲͲˆ‘”͵ͳƒ†…ƒ•’ƒ•‡Ǧ͵‘˜‡”‹‰Š–ƒ–ͶιǤ‡ŽŽ•™‡”‡ˆ—”–Š‡”‹…—„ƒ–‡†™‹–ŠŽ‡šƒǦͶͺͺƒ–‹Ǧ ”ƒ„„‹– ƒ† Ž‡šƒǦͷͻͶ ƒ–‹Ǧ‘—•‡ ȋ ˜‹–”‘‰‡ǡ ƒ”Ž•”—Š‡ǡ ‡”ƒ›Ȍǡ ”‡•’‡…–‹˜‡Ž›Ǥ ‘” ‡‰ƒ–‹˜‡ …‘–”‘Ž•ǡ …‡ŽŽ• ™‡”‡ –”‡ƒ–‡† ‘‹––‹‰ –Š‡ ϐ‹”•– ƒ–‹„‘†›Ǥ ƒ‰‡• ™‡”‡ –ƒ‡ ™‹–Š –Š‡ Š‡Ž’ ‘ˆ ƒ ‡‹•• š‹‘˜‡”– ʹͲͲͲ ‹…”‘•…‘’‡‡“—‹’’‡†™‹–Šƒš‹‘…ƒ…ƒ‡”ƒƒ†’Š‘–‘•™‡”‡ƒ…“—‹”‡†™‹–ŠƒͶͲšŽƒǦ’‘…Š”‘ƒ– ‘„Œ‡…–‹˜‡Ǥ ‘”–Š‡ƒƒŽ›•‹•‘ˆ…ƒ•’ƒ•‡Ǧ͵•‹‰ƒŽ‹–‡•‹–›ǡ–Š‡ ƒ‰‡ ƒƒŽ›•‹••‘ˆ–™ƒ”‡ȋƒ–‹‘ƒŽ •–‹–—–‡•‘ˆ Health (NIHȌǡ‡–Š‡•†ƒǡȌ™ƒ•—•‡†Ǥ

Statistical analysis •–ƒ–‹•–‹…ƒŽƒƒŽ›•‡•†ƒ–ƒƒ”‡’”‡•‡–‡†ƒ•–Š‡‡ƒάǤƒ–ƒ™‡”‡ƒƒŽ›œ‡†„›—•‹‰ ”ƒ’Šƒ† –ƒ–͵•‘ˆ–™ƒ”‡ˆ‘”‹†‘™•ȋƒ‹‡‰‘ǡǡȌƒ†–Š‡•–ƒ–‹•–‹…ƒŽ•‹‰‹ϐ‹…ƒ…‡™ƒ•‡•–‹ƒ–‡†„› ™‹–Š —‡›Ǧ”ƒ‡” —Ž–‹’Ž‡ …‘’ƒ”‹•‘ ’‘•– –‡•–Ǥ ƒŽ—‡• ‘ˆ ’δͲǤͲͷ ™‡”‡ …‘•‹†‡”‡† –‘ „‡ •–ƒ–‹•–‹…ƒŽŽ› •‹‰‹ϐ‹…ƒ–Ǥ

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Nitric oxide production ‡™ƒ›–‘‹˜‡•–‹‰ƒ–‡’”‘†—…–‹‘‹„‹‘Ž‘‰‹…ƒŽ•›•–‡•‹•–‘‡ƒ•—”‡‹–”‹–‡ǡ™Š‹…Š‹•–Š‡•–ƒ„Ž‡ ƒ† ‘Ǧ˜‘Žƒ–‹Ž‡ „”‡ƒ†‘™ ’”‘†—…– ‘ˆ Ǥ ‘” –Š‹•ǡ ”‹‡•• ”‡ƒ‰‡– •›•–‡ ™ƒ• ‡’Ž‘›‡† ȋ”‘‡‰ƒǡ

‡”ƒ›ȌǤ ”‹‡ϐŽ›ǡ ͷͲρŽ ‘ˆ …—Ž–—”‡ •—’‡”ƒ–ƒ– ™ƒ• ‹…—„ƒ–‡† ™‹–Š ƒ ‡“—ƒŽ ˜‘Ž—‡ ‘ˆ ”‹‡•• ”‡ƒ‰‡– ȋͲǤͷΨ •—Žˆƒ‹Žƒ‹†‡ǡ ͲǤͲͷΨ NǦȋͳǦƒ’Š–Š›ŽȌǦ‡–Š›Ž‡‡†‹ƒ‹‡ †‹Š›†”‘…ŠŽ‘”‹†‡ ‹ ʹǤͷΨ ͵ͶȌ ‹ ͻ͸Ǧ™‡ŽŽ –‹••—‡…—Ž–—”‡’Žƒ–‡ˆ‘”ͳͲ‹—–‡•ƒ–”‘‘–‡’‡”ƒ–—”‡ǤŠ‡ƒ„•‘”„ƒ…‡ƒ–ͷͷͲ™ƒ•‡ƒ•—”‡†™‹–Š ƒ ‹…”‘’Žƒ–‡ ”‡ƒ†‡” ȋ‹ƒ” ‹…”‘’Žƒ–‡ „•‘”„ƒ…‡ ‡ƒ†‡”ǡ ‹‘Ǧƒ†ǡ ò‹…Šǡ ‡”ƒ›Ȍ ‹…Ž—†‹‰ ‹–”‹–‡ •–ƒ†ƒ”†•Ǥ

Cellular Physiology and Biochemistry

DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb



Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

SWR/J

A

C57BL/6

a

b

c

d

e

f

6 d p.i.

9 d p.i.

16 d p.i.

2 months p.i.

Prussian Blue

CVB3 VP1

B

Prussian Blue

C 6,00E+08

4,00E+08 CVB3 3,00E+08

CVB3+Fe

2,00E+08

*

1,40E+07

**

5,00E+08

viral particles

viral transcripts

Fig. 1. …”‡ƒ•‡†‹”‘ deposition in suscep–‹„Ž‡Ȁ ‹…‡ƒ† its effect over viral replication in vitro. Ǥ —•…‡’–‹„Ž‡Ȁ ȋƒǡ …ǡ‡Ȍ„—–‘–”‡•‹•–ƒ– ͷ͹Ȁ͸ȋ„ǡ†ǡˆȌ‹…‡ show iron deposits ȋ…Ȍ ™‹–Š‹ …‘š•ƒ…‹evirus-infected myo…›–‡• ȋ‡ǡ ͵ ͳ ’”‘–‡‹ȌǤǤ …”‡ƒ•‡† iron deposition in macrophages (ar”‘™•Ȍƒ†…ƒ”†‹‘›‘cytes of susceptible ͵Ǧ‹ˆ‡…–‡† Ȁ  mice in the course of ‹ˆ‡…–‹‘Ǥ  Ǥ —ƒ–‹ϐ‹…ƒ–‹‘ ‘ˆ ‹…”‡ƒ•‡† ‡–‡”‘˜‹”—•  ȋ“ǦȌ ƒ† ‹ˆ‡…tious virions (plaque ƒ••ƒ›Ȍƒˆ–‡”‹”‘–”‡ƒ–‡–‹‡ƒ…‡ŽŽ•Ǣ”‡sults represent mean ά  ‘ˆ –Š”‡‡ ‹†‡pendent experiments (ȗȗ’δͲǤͲͳǡ ȗ’δͲǤͲͷȌǤ …ƒŽ‡„ƒ”ǣ…ǡ†ǡ‡ǡˆͳͲͲ ρǢ͸†’‹ͳͲͲρǡͻ† ’‹ ʹͲͲ Pǡ ͳ͸ † ’‹ǡ ʹ ‘–Š•’‹͵ͲρǤ

Cell Physiol Biochem 2014;33:52-66

1,20E+07 1,00E+07 8,00E+06 6,00E+06 4,00E+06

1,00E+08

2,00E+06 0,00E+00

0,00E+00

0h

8h

CVB3 8h

CVB3 + Fe 8h

Increased iron deposition in hearts of CVB3-infected SWR/J mice and the effect of iron on viral replication ‡Šƒ˜‡’”‡˜‹‘—•Ž›•Š‘™–Šƒ–Ȁ ‹…‡„—–‘–ͷ͹Ȁ͸‹…‡ƒ”‡•—•…‡’–‹„Ž‡ˆ‘” –Š‡†‡˜‡Ž‘’‡–‘ˆ…Š”‘‹…͵›‘…ƒ”†‹–‹•™Š‹…Š‹•ƒ••‘…‹ƒ–‡†™‹–Š•‡˜‡”‡”‡‘†‡ŽŽ‹‰ ’”‘…‡••‡•ƒ–Žƒ–‡”•–ƒ‰‡•‘ˆ‹ˆ‡…–‹‘ȏͳͶȐǤ –‡”‡•–‹‰Ž›ǡ‹–Š‹••–—†›™‡ˆ‘—†–Šƒ–‹–Š‡ ‹ˆ‡…–‡†ƒ††ƒƒ‰‡†›‘…ƒ”†‹—‘ˆȀ ‹…‡ȋ ‹‰Ǥͳǡ”‹…Š”‘‡Ȍ–Š‡‹”‘†‡’‘•‹–•ƒ”‡ ’”‡•‡–™‹–Š‹‹ˆ‡…–‡†›‘…›–‡•ǡƒ••Š‘™„›”—••‹ƒ„Ž—‡ȋ…Ȍƒ†‹—‘Š‹•–‘…Š‡‹…ƒŽ ͵˜‹”ƒŽ’”‘–‡‹ǦͳȋͳȌ•–ƒ‹‹‰•ȋ‡Ȍ‹…‘””‡•’‘†‹‰–‹••—‡•‡…–‹‘•Ǥ …‘–”ƒ•–ǡ͵Ǧ ‹ˆ‡…–‡†ͷ͹Ȁ͸‹…‡ǡ™Š‹…Šƒ”‡”‡•‹•–ƒ––‘…Š”‘‹…›‘…ƒ”†‹–‹•ǡ†‹†‘–”‡˜‡ƒŽ•‹‰‹ϐ‹…ƒ– …ƒ”†‹ƒ…†ƒƒ‰‡ȋ„Ȍǡ‹”‘†‡’‘•‹–•ȋ†Ȍ‘”’‡”•‹•–‡–˜‹”—•‹ˆ‡…–‹‘ȋˆȌǤ‡ƒŽ•‘ˆ‘—†–Šƒ–‹ –Š‡…‘—”•‡‘ˆ…Š”‘‹…›‘…ƒ”†‹–‹•‹Ȁ ‹…‡ǡ‹”‘‹•‘–‘Ž›‹…‘”’‘”ƒ–‡†‹†ƒƒ‰‡† ›‘…›–‡•„—–ƒŽ•‘‹ƒ…”‘’Šƒ‰‡•‹•‹‰‹ϐ‹…ƒ–ƒ‘—–••–ƒ”–‹‰ƒ–ͳ͸†ƒ›•’Ǥ‹Ǥȋ ‹‰ǤͳȌǤ  ›‘…›–‡•ǡ‹”‘ƒ……——Žƒ–‹‘™ƒ•†‡–‡…–ƒ„Ž‡ƒŽ”‡ƒ†›ˆ”‘†ƒ›͸’Ǥ‹Ǥ‘Ǥ –‡”‡•–‹‰Ž›ǡ‹–Š‡

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Results

Cellular Physiology and Biochemistry

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

57

Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

DHE

A

© 2014 S. Karger AG, Basel www.karger.com/cpb

overlay

VP1

d0

d16

DHE

B

Control

CVB3

Fe

Fig. 2.š‹†ƒ–‹˜‡•–”‡••‹‹”‘ǦŽ‘ƒ†‡†͵Ǧ‹ˆ‡…–‡†Š‡ƒ”–•‘ˆȀ ‹…‡ǤǤ—’‡”‘š‹†‡™Š‹…Š‹•”‡Ž‡ƒ•‡† ˆ”‘‹ϐ‹Ž–”ƒ–‹‰ƒ…”‘’Šƒ‰‡•ȋ™Š‹–‡ƒ””‘™•Ȍƒ††‡–‡…–‡†„›•–ƒ‹‹‰™‹–Š–Š‡•—’‡”‘š‹†‡Ǧ•‡•‹–‹˜‡†›‡ †‹Š›†”‘‡–Š‹†‹—  ȋ”‡† ϐŽ—‘”‡•…‡…‡Ȍǡ •Š‘™• ‹…”‡ƒ•‡† •‹‰ƒŽ ‹–‡•‹–› ‹ …Ž‘•‡ ˜‹…‹‹–› ‘ˆ ‹ˆ‡…–‡† ›‘…›–‡•ȋͳ͸†ƒ›•’Ǥ‹Ǣ…‘Ǧ•–ƒ‹‹‰™‹–Š͵ͳȋ‰”‡‡Ȍƒ† ȌǤǤš‹†ƒ–‹˜‡•–”‡••ȋ•–ƒ‹‹‰Ȍ‹ ͵ƒ†‹”‘–”‡ƒ–‡†ʹ͸ͶǤ͹ƒ…”‘’Šƒ‰‡•Ǥ…ƒŽ‡„ƒ”ǣʹͲͲρǤ

Oxidative stress is induced by CVB3 replication and iron ‘”†‡”–‘‰ƒ‹‘”‡‹•‹‰Š–‹–‘–Š‡‡ˆˆ‡…–•‘ˆ‹”‘‹–Š‡˜‹”—•Ǧ‹ˆ‡…–‡†›‘…ƒ”†‹—ǡ ™‡ ‹˜‡•–‹‰ƒ–‡† –Š‡ •–”‡•• …‘†‹–‹‘• ‹ –Š‡•‡ Š‡ƒ”–•Ǥ ͵ ›‘…ƒ”†‹–‹• ™ƒ• ˆ‘—† –‘ „‡ associated with increased oxidative stress mainly within infected areas, as visualized „› …‘Ž‘…ƒŽ‹œƒ–‹‘ ‘ˆ ͵ ͳ ‹ ›‘…›–‡•ǡ ™‹–Š ‹ϐ‹Ž–”ƒ–‹‰ ƒ…”‘’Šƒ‰‡• ƒ• •‘—”…‡ ‘ˆ •—’‡”‘š‹†‡ȋ ‹‰Ǥʹǡ™Š‹–‡ƒ””‘™•ǡͳ͸†ƒ›•’‹ǤȌǤ …‘–”ƒ•–ǡ”‡•‹•–ƒ–ͷ͹Ȁ͸‹…‡”‡˜‡ƒŽ‡† Ž‡••‘š‹†ƒ–‹˜‡•–”‡••™Š‹…Š…‘””‡Žƒ–‡•™‹–ŠŽ‘™‹—‡…‡ŽŽƒ…–‹˜‹–›ƒˆ–‡”͵‹ˆ‡…–‹‘ȋ†ƒ–ƒ ‘–•Š‘™ȌǤ —”–Š‡”‹ˆ‘”ƒ–‹‘‹•’”‘˜‹†‡†‹‹”‘ǦŽ‘ƒ†‡†ƒ†˜‹”—•Ǧ•–‹—Žƒ–‡†ʹ͸ͶǤ͹ ƒ…”‘’Šƒ‰‡•ǡ”‡˜‡ƒŽ‹‰ƒ—’”‡‰—Žƒ–‹‘‘ˆ•—’‡”‘š‹†‡’”‘†—…–‹‘—†‡”„‘–Š…‘†‹–‹‘•Ǥ• ‡š’‡…–‡†ǡ”‡’Ž‹…ƒ–‹‰͵™ƒ•ˆ‘—†–‘•–”‡••–Š‡…‡ŽŽ•–‘ƒŠ‹‰Š‡”‡š–‡––Šƒ‹”‘ȋ ‹‰ǤʹȌǤ —Ž–‹˜ƒ–‡†—–”‡ƒ–‡†ƒ…”‘’Šƒ‰‡•™‡”‡ˆ‘—†–‘„‡‡‰ƒ–‹˜‡ˆ‘”•—’‡”‘š‹†‡‡š’”‡••‹‘Ǥ

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…‘—”•‡‘ˆ›‘…ƒ”†‹–‹•–Š‡ƒŒ‘”‹–›‘ˆ‹”‘†‡’‘•‹–•™‡”‡‘„•‡”˜‡†‹›‘…›–‡•Ž‘…ƒ–‡†ƒ––Š‡ „‘”†‡”–‘—‹ˆ‡…–‡†–‹••—‡ȋͻ†ƒ›•’Ǥ‹ȌǤ˜‡ƒˆ–‡”–Š‡†‡…Ž‹‡‘ˆ‹ϐŽƒƒ–‹‘—‡”‘—• damaged cardiomyocytes were found to be heavily loaded with iron, as exemplarily shown ‹ƒŠ‡ƒ”–ʹ‘–Š•’Ǥ‹ǤǤ‘‰ƒ‹‘”‡‹•‹‰Š–‹–‘–Š‡‹–‡”ƒ…–‹‘‘ˆ‹”‘ƒ†˜‹”ƒŽ”‡’Ž‹…ƒ–‹‘ǡ ™‡“—ƒ–‹ϐ‹‡†–Š‡‡–‡”‘˜‹”ƒŽ„›“Ǧ‹—–”‡ƒ–‡†ƒ†‹”‘Ǧ–”‡ƒ–‡†ȋͳͲͲρ‘ŽȀȌ ͵Ǧ‹ˆ‡…–‡† ‡ƒ …‡ŽŽ•ǡ ƒ ™‡ŽŽ †‡ϐ‹‡† …‡ŽŽ …—Ž–—”‡ ‘†‡Ž ‘ˆ ͵ ›‘…ƒ”†‹–‹•Ǥ ”‘Ǧ –”‡ƒ–‡†•ƒ’Ž‡•”‡˜‡ƒŽ‡†ƒͶǦˆ‘Ž†‹…”‡ƒ•‡‹–Š‡—„‡”‘ˆ˜‹”ƒŽ–”ƒ•…”‹’–•…‘’ƒ”‡† –‘—–”‡ƒ–‡†‹ˆ‡…–‡†…‡ŽŽ•ȋ ‹‰ǤͳȌǤ‘””‡•’‘†‡––‘–Š‡”‡•—Ž–•‹“Ǧǡ™‡ˆ‘—†‹ ’Žƒ“—‡ƒ••ƒ›‡š’‡”‹‡–•ƒ•‹‰‹ϐ‹…ƒ–‹…”‡ƒ•‡‘ˆ‹ˆ‡…–‹‘—•˜‹”‹‘•‹‡ƒ…‡ŽŽ•‹’”‡•‡…‡ ‘ˆ‹”‘ȋ ‹‰ǤͳȌǤ

Cellular Physiology and Biochemistry Fig. 3. ‹  ƒ† ’”‘–‡‹ †‡–‡…–‹‘ ‹ ͵Ǧ‹ˆ‡…–‡† Š‡ƒ”–• ƒ† …—Ž–‹˜ƒ–‡† ƒ…”‘’Šƒ‰‡•Ǥ Ǥ –”‘‰ ‹  ‡š’”‡••‹‘ „› ‹ϐ‹Ž–”ƒ–‹‰ ƒ…”‘’Šƒ‰‡• ‹ –Š‡ Š‡ƒ”– ‘ˆ ƒ…—–‡Ž› ͵Ǧ‹ˆ‡…–‡† Ȁ ‹…‡•Š‘™„›”ƒ†‹‘ƒ…–‹˜‡ ȋͺ†ƒ›•’Ǥ‹Ǥǡ•…ƒŽ‡„ƒ”ǣͳͲͲρȌǤ Ǥ‡’”‡•‡–ƒ–‹˜‡‹—‘„Ž‘–ˆ‘” ‹’”‘–‡‹‡š’”‡••‹‘‹Ȁ  heart extracts revealing highest Ž‡˜‡Ž•†—”‹‰ƒ…—–‡›‘…ƒ”†‹–‹•ȋͺ †ƒ›• ’Ǥ‹ǤȌǤ Ǥ ‹ ’”‘–‡‹ ‡š’”‡••‹‘‹‹”‘‘”͵Ǧ–”‡ƒ–‡† ʹ͸ͶǤ͹ƒ…”‘’Šƒ‰‡•ͺŠ‘—”•’Ǥ‹Ǥǡƒ• •Š‘™„› Ǥ…ƒŽ‡„ƒ”ǣʹͲͲρǤ

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb

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A

iNOS mRNA d8

iNOS mRNA d0

SWR/J

C57BL/6

B iNOS ȕ-tubulin

d0

d4

d8

d16

d28

C

Fe

CVB3

Induction of iNOS by iron and CVB3 ’”‡‰—Žƒ–‹‘‘ˆ–Š‡‹‹•‘™–‘„‡ƒƒŒ‘”…ƒ—•‡‘ˆ‘š‹†ƒ–‹˜‡•–”‡••‹–Š‡Š‡ƒ”–Ǥ ‡ Šƒ˜‡ ’”‡˜‹‘—•Ž› •Š‘™ –Šƒ– ‹ ‡š’”‡••‹‘ ‹ …ƒ”†‹ƒ… ƒ…”‘’Šƒ‰‡• …‘–”‹„—–‡• –‘ –Š‡‹—‘’ƒ–Š‘Ž‘‰›‘ˆ‘‰‘‹‰›‘…ƒ”†‹–‹•‹•—•…‡’–‹„Ž‡Ȁ ‹…‡ȏͷȐǤ ‘”†‡”–‘ ‡˜ƒŽ—ƒ–‡™Š‡–Š‡”‹”‘‹ϐŽ—‡…‡•‹‡š’”‡••‹‘‹–Š‡…‘—”•‡‘ˆ͵›‘…ƒ”†‹–‹•ǡŠ‡ƒ”–• ‘ˆ‹”‘ǦŽ‘ƒ†‡†Ȁ ‹…‡™‡”‡ƒƒŽ›œ‡†‹–Š‡…‘—”•‡‘ˆ›‘…ƒ”†‹–‹•Ǥ …‘–”ƒ•––‘”‡•‹•–ƒ– ͷ͹Ȁ͸‹…‡ǡȀ ‹…‡”‡˜‡ƒŽ—‡”‘—•‹’‘•‹–‹˜‡…‡ŽŽ•‹ƒ”‡ƒ•‘ˆ•‡˜‡”‡ ‹ϐŽƒƒ–‹‘ˆ‘ŽŽ‘™‹‰͵‹ˆ‡…–‹‘ȋ ‹‰Ǥ͵ȌǤ‘””‡•’‘†‡––‘–Š‡‹•‹–—Š›„”‹†‹œƒ–‹‘ ”‡•—Ž–•ǡ–Š‡ƒš‹—‘ˆ‹’”‘–‡‹‡š’”‡••‹‘™ƒ•‘„•‡”˜‡†ͺ†ƒ›•’‹Ǥȋ ‹‰Ǥ͵ȌǤ ‘”†‡” –‘‡˜ƒŽ—ƒ–‡–Š‡‡ˆˆ‡…–‘ˆ‹”‘ƒ†͵‹ˆ‡…–‹‘‘‹‡š’”‡••‹‘in vitroǡ…—Ž–‹˜ƒ–‡† ʹ͸ͶǤ͹ƒ…”‘’Šƒ‰‡•™‡”‡‹˜‡•–‹‰ƒ–‡†„›‹—‘ϐŽ—‘”‡•…‡…‡•–ƒ‹‹‰•Ǥ‘””‡•’‘†‹‰–‘ ‘—”‘„•‡”˜ƒ–‹‘•‘ˆ•—’‡”‘š‹†‡’”‘†—…–‹‘ǡ™‡ˆ‘—†ƒ—’”‡‰—Žƒ–‹‘‘ˆ‹’”‘–‡‹Ž‡˜‡Ž• ‹„‘–Šǡ͵Ǧ•–‹—Žƒ–‡†ƒ•™‡ŽŽƒ•‹‹”‘–”‡ƒ–‡†ʹ͸ͶǤ͹ƒ…”‘’Šƒ‰‡•ǡ™Š‡…‘’ƒ”‡† –‘—–”‡ƒ–‡†…‘–”‘Ž…‡ŽŽ•ȋ ‹‰Ǥ͵ȌǤ

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control

Cellular Physiology and Biochemistry

DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb

59

Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

AA SWR/J

a

b

c HO-1 mRNA d8

HO-1 mRNA d0

HO-1 protein d8

C57BL/6

d

f

e

BB Control

Fe

Fe CVB3

CVB3

Fe CVB3

HO1

overlay

VP1

C

C relative values

Fig. 4.Ǧͳƒ†’”‘–‡‹ †‡–‡…–‹‘ ‹ ͵Ǧ‹fected hearts and cultivated …‡ŽŽ•Ǥ Ǥ ‡–‡…–‹‘ ‘ˆ Ǧͳ  „› ‹ •‹–— hybridiza–‹‘ ȋƒǡ „ǡ † ƒ† ‡Ȍ ƒ† ’”‘tein by immunohistochem‹•–”› ȋ… ƒ† ˆȌ ™‹–Š‹ ƒ…”‘’Šƒ‰‡• ȋƒ””‘™•Ȍ ƒ† …ƒ”†‹‘›‘…›–‡• ȋƒ””‘™Š‡ƒ†Ȍ ‹ hearts of infected susceptible and resistant mice, revealing Š‹‰ŠǦͳƒ†’”‘–‡‹ expression in susceptible Ȁ  ‹…‡ †—”‹‰ ƒ…—–‡ ͵ ›‘…ƒ”†‹–‹• ȋͺ †ƒ›• ’Ǥ‹ǤȌǤ…ƒŽ‡„ƒ”ǣͳͲͲρȋƒǦˆȌǤ Ǥ Ǧͳ  ‹ ͵Ǧ‹ˆ‡…–‡† ƒ†Ȁ‘” ‹”‘Ǧ–”‡ƒ–‡† ‡ƒ cells show increased protein ‡š’”‡••‹‘ͺŠ‘—”•’Ǥ‹‹–Š‡ —…Ž‡—•ȋ͵Ȍ‘”…›–‘’Žƒ• ȋ‹”‘Ȍ ‘” „‘–Š ȋ͵Ȁ‹”‘ȌǤ …ƒŽ‡ „ƒ”ǣ ʹͲ ρǤ Ǥ Ǧͳ representative immunoblot and densitometric analyses of three independent experi‡–•”‡˜‡ƒŽ‹…”‡ƒ•‡†Ǧͳ ’”‘–‡‹ ‡š’”‡••‹‘ ‹  ʹ͸ͶǤ͹Ǧ‡ƒ …‡ŽŽ …‘…—Ž–—”‡• ͺ Š‘—”• ’Ǥ‹Ǥ ‹ ’”‡•‡…‡ ‘ˆ ‹”‘ǡ•Š‘™‹‰–Šƒ–͵ƒ† iron potentiate each others ƒ…–‹˜‹–› ȋ•‹‰‹ϐ‹…ƒ…‡ ˜•Ǥ —treated cocultured control …‡ŽŽ•ǣ ȗ’δͲǤͲͷǡ ȗȗ’δͲǤͲͳ ƒ† ȗȗȗ’δͲǤͲͲͳȌǤ

Cell Physiol Biochem 2014;33:52-66

***

13 12 11 10 9 8 7 6 5 4 3 2 1 0

** *

control

Fe 8h

CVB3 8h Fe+CVB3 8h

HO-1

Heme-oxygenase 1 is upregulated by iron and CVB3 ‹–”‹…‘š‹†‡‹•‘™ƒ•‘‡‘ˆ–Š‡‘•–’‘–‡–‹†—…‡”•‘ˆǦͳ‰‡‡‡š’”‡••‹‘‹ various tissues in vivoȏͳͷȐƒ•™‡ŽŽƒ•‹†‹ˆˆ‡”‡–…‡ŽŽ–›’‡•in vitroȏͳ͸ȐǤ ‘”†‡”–‘‡˜ƒŽ—ƒ–‡ ™Š‡–Š‡” –Š‡ ‘š‹†ƒ–‹˜‡ •–”‡•• ”‡ƒ…–‹‘• ƒ› ‹ϐŽ—‡…‡ Ǧͳ ‡š’”‡••‹‘ in vivo, ™‡ ϐ‹”•– ‹˜‡•–‹‰ƒ–‡† Š‡ƒ”–• ‘ˆ ͵Ǧ‹ˆ‡…–‡† Ȁ  ƒ† ͷ͹Ȁ͸ ‹…‡Ǥ —•…‡’–‹„Ž‡ Ȁ  ‹…‡

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ȕ-tubulin

Cellular Physiology and Biochemistry A

C

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb



Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

B

CVB

1mM L-NAME CVB

Fe

1mM L-NAME Fe

Fe CVB

1mM L-NAME Fe CVB

Control

continued

Ǧͷ‡š’”‡••‹‘‹•‹ϔŽ—‡…‡†„›‹ƒ…–‹˜‹–›ƒ†‡†‹ƒ–‡•‘š‹†ƒ–‹˜‡•–”‡•• • ‡–‹‘‡† „‡ˆ‘”‡ǡ •—•–ƒ‹‡† ‹ ƒ…–‹˜‹–› Šƒ• „‡‡ ’”‘˜‡ –‘ ”‡•—Ž– ‹ ‘‰‘‹‰ …ƒ”†‹ƒ…‹Œ—”›‹͵›‘…ƒ”†‹–‹•Ǥ•‹–‹•™‡ŽŽ‘™–Šƒ–…Šƒ‰‡•‹‹’”‘–‡‹•–”‘‰Ž›

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”‡˜‡ƒŽ…‘’ƒ”ƒ„Ž›Š‹‰Š‡”ƒ‘—–•‘ˆǦͳƒ†’”‘–‡‹‡š’”‡••‹‰‹—‡…‡ŽŽ•‹ …ƒ”†‹ƒ…‹ϐŽƒƒ–‘”›Ž‡•‹‘•…‘’ƒ”‡†–‘ͷ͹Ȁ͸‹…‡ƒ–ƒ›–‹‡‘ˆ‹ˆ‡…–‹‘ȋ†ƒ–ƒ‘– •Š‘™Ȍǡ”‡ƒ…Š‹‰ƒƒš‹—†—”‹‰–Š‡ƒ…—–‡’Šƒ•‡‘ˆ–Š‡†‹•‡ƒ•‡ȋͺ†ƒ›•’Ǥ‹Ǥǡ ‹‰ǤͶȌǤ  †‡–ƒ‹Ž‡† ‹—‘Š‹•–‘…Š‡‹…ƒŽ •–—†› ƒ– –Š‡ …‡ŽŽ—Žƒ” Ž‡˜‡Ž ”‡˜‡ƒŽ‡† –Šƒ– ‹…”‡ƒ•‡† Ǧͳ’”‘–‡‹‡š’”‡••‹‘‹•ƒ‹Ž›‘„•‡”˜‡†‹ƒ…”‘’Šƒ‰‡•ȋƒ””‘™•Ȍ„—–‹•ƒŽ•‘†‡–‡…–‡†‹ •‹‰Ž‡…ƒ”†‹‘›‘…›–‡•ȋƒ””‘™Š‡ƒ†ȌǤ ‘”†‡”–‘‹˜‡•–‹‰ƒ–‡–Š‡‡ˆˆ‡…–•‘ˆ‹”‘ƒ†͵‘Ǧͳ‡š’”‡••‹‘in vitroǡ‡ƒ…‡ŽŽ• ™‡”‡–”‡ƒ–‡†„›‡‹–Š‡”‹”‘ƒ†Ȁ‘”͵Ǥ•†‡‘•–”ƒ–‡†‹ ‹‰—”‡Ͷǡ‹—‘ϐŽ—‘”‡•…‡…‡ ‘ˆ—–”‡ƒ–‡†…‘–”‘Ž…‡ŽŽ•‹ŽŽ—•–”ƒ–‡•–Š‡’”‡•‡…‡‘ˆǦͳ‹—…Ž‡‘Ž‹ǡƒϐ‹†‹‰™Š‹…Š™ƒ• ’”‡˜‹‘—•Ž›”‡’‘”–‡†„›‹‘Ž–‹‹…—Ž–—”‡†ƒ•–”‘…›–‡•ȏͳ͹ȐǤ …—„ƒ–‹‘‘ˆ…‡ŽŽ•™‹–Š‹”‘ƒ†Ȁ ‘”͵”‡•—Ž–‡†‹–Š‡—’”‡‰—Žƒ–‹‘‘ˆǦͳ’”‘–‡‹‹†‹ˆˆ‡”‡–…‘’ƒ”–‡–•‘ˆ–Š‡…‡ŽŽ• ƒ†”‡˜‡ƒŽ‡†ƒ•’”‡ƒ†‘ˆǦͳ™‹–Š‹–Š‡—…Ž‡—•ȋ͵Ȍƒ•™‡ŽŽƒ•‘˜‡”–Š‡…›–‘’Žƒ•ȋ‹”‘Ȍ ‘”„‘–Šȋ͵Ȁ‹”‘ȌǤ‘•‹—Žƒ–‡–Š‡‹–‡”ƒ…–‹‘‘ˆ›‘…›–‡•ƒ†ƒ…”‘’Šƒ‰‡•‘„•‡”˜‡†in vivo ™‹–Š ”‡‰ƒ”† –‘ Ǧͳ ‡š’”‡••‹‘ǡ ™‡ ’‡”ˆ‘”‡† ™‡•–‡” „Ž‘– ƒƒŽ›•‡• ‘ˆ …›–‘’Žƒ•‹… ’”‘–‡‹ ‡š–”ƒ…–• ‘„–ƒ‹‡† ˆ”‘  ʹ͸ͶǤ͹ Ȃ‡ƒ …‡ŽŽ …‘…—Ž–—”‡• ȋ ‹‰Ǥ ͶȌǤ ‘’ƒ”‡† –‘ —–”‡ƒ–‡† …‡ŽŽ•ǡ ƒ •‹‰‹ϐ‹…ƒ– ’‘–‡–‹ƒ–‹‘ ‹ Ǧͳ ’”‘–‡‹ ‡š’”‡••‹‘ ™ƒ• ‘„•‡”˜‡† ƒˆ–‡” ƒ††‹–‹‘‘ˆ͵ȋ’δͲǤͲͷȌǡ‹”‘ȋ’δͲǤͲͳȌ‘”‹”‘Ȁ͵ȋ’δͲǤͲͲͳȌǤ

Cellular Physiology and Biochemistry

DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb



Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

D

…‘””‡Žƒ–‡ ™‹–Š …Šƒ‰‡• ‹ ‹–”‹–‡•ǡ ™‡ ‡ƒ•—”‡† ‹–”‹–‡ Ž‡˜‡Ž• ‹  ʹ͸ͶǤ͹Ȁ‡ƒ …‡ŽŽ …‘…—Ž–—”‡•ƒ†ˆ‘—†ǡƒ•‡š’‡…–‡†ǡƒ—’”‡‰—Žƒ–‹‘‘ˆ‹–”‹–‡Ž‡˜‡Ž•—’‘‹”‘‘”Ȁƒ†͵ ‹ˆ‡…–‹‘Ǥ …—„ƒ–‹‘ ™‹–Š Ǧ ”‡•—Ž–‡† ‹ •‹‰‹ϐ‹…ƒ–Ž› †‡…”‡ƒ•‡† Ž‡˜‡Ž• ‘ˆ ‹–”‹–‡ ‹ ͵ƒ†͵Ȁ‹”‘–”‡ƒ–‡†…‡ŽŽ•ȋ ‹‰ǤͷȌǤ ‘”†‡”–‘‹˜‡•–‹‰ƒ–‡™Š‡–Š‡”Ǧͳ‹•”‡‰—Žƒ–‡† ˜‹ƒ ‹ǡ ™‡ „Ž‘…‡† ‹ ƒ…–‹˜‹–› ‹ Ǧ –”‡ƒ–‡†  ʹ͸ͶǤ͹ Ȃ‡ƒ …‡ŽŽ …‘…—Ž–—”‡•Ǥ ‡•–‡” „Ž‘– ƒƒŽ›•‹• ”‡˜‡ƒŽ‡† –Šƒ– ‹Š‹„‹–‹‘ ‘ˆ ‹ ƒ…–‹˜‹–› „› Ǧ •‹‰‹ϐ‹…ƒ–Ž› ”‡†—…‡•Ǧͳ’”‘–‡‹‡š’”‡••‹‘‹͵ǡ‹”‘ƒ†͵Ȁ‹”‘–”‡ƒ–‡†…‡ŽŽ•ȋȋ’δͲǤͲͲͳȌ ‹‰Ǥ ͷȌǤŠ‹•‘„•‡”˜ƒ–‹‘‹••—’’‘”–‡†„›‹—‘ϐŽ—‘”‡•…‡…‡’”‘˜‹‰ƒ”‡†—…–‹‘‹–Š‡Ǧͳ •‹‰ƒŽ‹–‡•‹–›—’‘„Ž‘…‹‰‘ˆ‹„›Ǧˆ‘ŽŽ‘™‡†„›‹”‘‘”Ȁƒ†͵–”‡ƒ–‡– ȋ ‹‰ǤͷȌǡ‹†‹…ƒ–‹‰–Šƒ–‹‘†—Žƒ–‡•Ǧͳ‡š’”‡••‹‘Ǥ —’‡”‘š‹†‡ ’”‘†—…–‹‘ …‘–”‹„—–‡• –‘ ‘š‹†ƒ–‹˜‡ •–”‡•• ƒ† …ƒ „‡ ‘„•‡”˜‡† ‹ …‡ŽŽ• —’‘͵‹ˆ‡…–‹‘ƒ†–”‡ƒ–‡–™‹–Š‹”‘Ǥ ‘”†‡”–‘‡˜ƒŽ—ƒ–‡–Š‡…‘–”‹„—–‹‘‘ˆǦͳ ‡š’”‡••‹‘‘–Š‡‰‡‡”ƒ–‹‘‘ˆ•—’‡”‘š‹†‡ǡǦͳ™ƒ•„Ž‘…‡†„›Ǥ••Š‘™‹ ‹‰—”‡ ͷǡ™‡ˆ‘—†ƒ…Ž‡ƒ””‡†—…–‹‘‹•—’‡”‘š‹†‡’”‘†—…–‹‘ƒˆ–‡”Ǧͳ‹Š‹„‹–‹‘—’‘‹”‘‘”Ȁ

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Fig. 5. Ǧͳ ‹• ‘†—Žƒ–‡† „›  ’”‘†—…–‹‘ ƒ† ‹creases oxidative stress in Ǧʹ͸ͶǤ͹ ƒ…”‘’Šƒ‰‡•Ȁ ‡ƒ …‘…—Ž–—”‡•Ǥ Ǥ ‹–”‹–‡ production is decreased by Ǧ ‹ ƒ…”‘’Šƒ‰‡• which were treated with ͵ ȋȗȗȗ’δͲǤͲͲͳȌǡ ‹”‘ ȋ’ε ͲǤͲͷȌǡ ‘” ‹”‘ Ȁ͵ ȋȗ’δ ͲǤͲͳȌǤ Ǥ ‡…”‡ƒ•‡† Ǧͳ ’”‘–‡‹ ‡š’”‡••‹‘ ™Š‡  ’”‘†—…–‹‘ ‹• ‹Š‹„‹–‡† „› Ǧ ‹ immunoblots (densitometric analyses of three independent experiments ȋȗȗȗ’δ ͲǤͲͲͳȌǤ Ǥ  —‘ϐŽ—‘”‡•…‡…‡ •Š‘™• †‡…”‡ƒ•‡†Ǧͳ‡š’”‡••‹‘‹ ͵ǡ‹”‘ƒ†͵Ȁ‹”‘Ǧ treated cocultured cells in ’”‡•‡…‡ ‘ˆ ǦǤ …ƒŽ‡ „ƒ”ǣ ʹͲ ρǤ Ǥ —’‡”‘š‹†‡ production, as detected „›  •–ƒ‹‹‰ ȋ”‡†Ȍ ‹  ʹ͸ͶǤ͹ ƒ…”‘’Šƒ‰‡• ȋ‰”‡‡Ȍ ™Š‹…Š ™‡”‡ …‘…—Ž–—”‡† ™‹–Š ͵Ǧ‹ˆ‡…–‡† ‡ƒ …‡ŽŽ•ǡ ‹• †‡…”‡ƒ•‡† ‹ ’”‡•‡…‡‘ˆǦͳǦ†‘™”‡‰ulating tin mesoporphyrin ȋǡ ʹͲȌǤ ‡ˆ‘”‡ –Š‡ ‡š’‡”‹‡–ǡ  ʹ͸ͶǤ͹ macrophages were loaded ™‹–Š ‡ŽŽ”ƒ…‡” ‰”‡‡ ‹ order to differentiate them ˆ”‘ ‡ƒ …‡ŽŽ•Ǥ …ƒŽ‡ „ƒ”ǣ ͷͲρǤ

Cell Physiol Biochem 2014;33:52-66

Cellular Physiology and Biochemistry

a

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb



Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

b

Fig. 6.’‘’–‘•‹•ƒ†‹”‘‹…ƒ”†‹‘›‘…›–‡•ʹ‘–Š•’Ǥ‹Ǥ‘ˆ͵Ǧ‹ˆ‡…–‡†Ȁ ‹…‡Ǥ —‘Š‹•–‘…Š‡‹•–”›ˆ‘”ƒ…–‹˜ƒ–‡†…ƒ•’ƒ•‡Ǧ͵ȋƒȌ‹ƒ…ƒ”†‹‘›‘…›–‡ʹ‘–Š•’Ǥ‹Ǥ͵ˆ‘ŽŽ‘™‡†„›”—••‹ƒ„Ž—‡•–ƒ‹ ȋ„Ȍ•Š‘™•–Šƒ–‹”‘ǦŽ‘ƒ†‡†…ƒ”†‹‘›‘…›–‡•ƒ”‡—†‡”‰‘‹‰ƒ’‘’–‘•‹•Ǥ…ƒŽ‡„ƒ”ǣͳͲρǤ

ƒ†͵–”‡ƒ–‡–†‡‘•–”ƒ–‹‰–Šƒ–Ǧͳ‡š’”‡••‹‘ƒ……‘—–•ˆ‘”‘š‹†ƒ–‹˜‡ƒ…–‹˜‹–‹‡•‹ ͵‹ˆ‡…–‹‘Ǥ HO-1 expression and apoptosis ‘”†‡”–‘‹˜‡•–‹‰ƒ–‡–Š‡…‘–”‹„—–‹‘‘ˆ͵’‡”•‹•–‡…‡ǡ‹…”‡ƒ•‡†‹”‘•–‘”ƒ‰‡ƒ• well as of oxidative stress on the survival of myocytes in vivo,™‡‹˜‡•–‹‰ƒ–‡†͵Ǧ‹ˆ‡…–‡† Š‡ƒ”–• ‘ˆ Ȁ  ‹…‡ „› Ž‹‰Š– ‹…”‘•…‘’›Ǥ • ‡š‡’Žƒ”‹Ž› †‡‘•–”ƒ–‡† ‹ ‹‰—”‡ ͸ǡ ™‡ ‘„•‡”˜‡†ƒ…–‹˜ƒ–‡†…ƒ•’ƒ•‡Ǧ͵‡š’”‡••‹‘ȋƒȌ‹–Š‘•‡…ƒ”†‹‘›‘…›–‡•™Š‹…Šƒ”‡ƒŽ•‘Ž‘ƒ†‡† ™‹–Š ‹”‘ ȋ„Ȍ ƒ– Žƒ–‡” •–ƒ‰‡• ‘ˆ –Š‡ †‹•‡ƒ•‡Ǥ Š‡•‡ ϐ‹†‹‰• ‹†‹…ƒ–‡ –Šƒ– ‹”‘ …‘–ƒ‹‹‰ ›‘…›–‡•—†‡”‰‘ƒ’‘’–‘•‹•‹’‡”•‹•–‡–Ž›͵Ǧ‹ˆ‡…–‡†‹…‡ǡ™Š‹…Š…‘””‡•’‘†•™‡ŽŽ–‘ ‘—”‘„•‡”˜ƒ–‹‘•‹Š—ƒ‹ϐŽƒƒ–‘”›‹”‘Ǧ‹†—…‡†…ƒ”†‹‘›‘’ƒ–Š›ȋ†ƒ–ƒ‘–•Š‘™ȌǤ ‘ ˆ—”–Š‡” †‡–‡”‹‡ ™Š‡–Š‡” –Š‡ Š‘•– …‡ŽŽ †‡ƒ–Š ƒ…Š‹‡”› ‹• ƒ…–‹˜ƒ–‡† „› ‹”‘ ƒ†͵‹ˆ‡…–‹‘in vitroǡ‡ƒ…‡ŽŽ•™‡”‡—•‡†ƒ•ƒ‘†‡Ž•›•–‡Ǥ•†‡‘•–”ƒ–‡†‹ ‹‰—”‡͹ǡ…ƒ•’ƒ•‡Ǧ͵™Š‹…Š™ƒ•ˆ‘—†–‘„‡ƒ…–‹˜ƒ–‡†„›‹”‘ǡŽ‘…ƒŽ‹œ‡•ƒ‹Ž›ƒ”‘—†–Š‡ —…Ž‡—•Ǥ Ž•‘ǡ ͵ ‹†—…‡† …ƒ•’ƒ•‡Ǧ͵ ƒ…–‹˜ƒ–‹‘ǡ Š‘™‡˜‡” ™‹–Š ƒ ’”‹ƒ”› Ž‘…ƒŽ‹œƒ–‹‘ ‹ –Š‡…›–‘’Žƒ•ǤŠ‡’”‡•‡…‡‘ˆ„‘–Š•–‹—Ž‹ǡ͵ƒ†‹”‘ǡ‹†—…‡•—…Ž‡ƒ”…‘†‡•ƒ–‹‘‹ ƒ††‹–‹‘–‘ƒ…–‹˜ƒ–‹‘‘ˆ…ƒ•’ƒ•‡Ǧ͵Ǥ ‘†‡Ž‹‡ƒ–‡–Š‡”‘Ž‡‘ˆǦͳ‹…ƒ•’ƒ•‡Ǧ͵ƒ…–‹˜ƒ–‹‘‘ˆ‹”‘Ǧƒ†͵Ǧ–”‡ƒ–‡†…‡ŽŽ•ǡ ™‡—•‡†ƒǦͳ•‹ƒ’’”‘ƒ…ŠǤ ’ƒ”ƒŽŽ‡Ž–‘–Š‡‡š’‡”‹‡–•’”‡•‡–‡†‹ ‹‰—”‡͹ǡ™‡ ’‡”ˆ‘”‡†‹—‘ϐŽ—‘”‡•…‡…‡•–ƒ‹‹‰ˆ‘”–Š‡†‡–‡…–‹‘‘ˆƒ…–‹˜ƒ–‡†…ƒ•’ƒ•‡Ǧ͵ǡŠ‘™‡˜‡”‹ ’”‡•‡…‡‘ˆǦͳ•‹ǤǦͳ•‹Ž‡…‹‰”‡•—Ž–‡†‹ƒ•‹‰‹ϐ‹…ƒ–”‡†—…–‹‘‘ˆ•‹‰ƒŽ‹–‡•‹–›‘ˆ …ƒ•’ƒ•‡Ǧ͵•‹‰ƒŽ•‹‹”‘ƒ†Ȁ‘”͵–”‡ƒ–‡†…‡ŽŽ•ȋ ‹‰Ǥ͹Ȍ…‘’ƒ”‡†–‘–Š‡…‘””‡•’‘†‡– ”‡•—Ž–• ‹ ‹‰Ǥ ͹Ǥ ƒ‰‡ ƒƒŽ›•‹• ‘ˆ …ƒ•’ƒ•‡Ǧ͵ •‹‰ƒŽ ‹–‡•‹–› ‹ ͵Ǧ‹ˆ‡…–‡† ƒ† ‹”‘Ǧ –”‡ƒ–‡†•ƒ’Ž‡•”‡˜‡ƒŽƒ•‹‰‹ϐ‹…ƒ–†‘™”‡‰—Žƒ–‹‘‘ˆ…ƒ•’ƒ•‡Ǧ͵‡š’”‡••‹‘‹’”‡•‡…‡‘ˆ Ǧͳ •‹ ȋ ‹‰Ǥ ͹Ȍ ȋ’δͲǤͲͲͳȌǤ Š‡•‡ ”‡•—Ž–• •—„•–ƒ–‹ƒ–‡ –Š‡ ”‘Ž‡ ‘ˆ Ǧͳ ‹ …ƒ•’ƒ•‡Ǧ͵ activation which is also observed in vivo ‹ ‹”‘ǦŽ‘ƒ†‡† ›‘…›–‡• ‘ˆ ͵Ǧ‹ˆ‡…–‡† ‹…‡ ȋ…‘’ƒ”‡ ‹‰Ǥ͸ȌǤ ‘”†‡”–‘˜‡”‹ˆ›–Š‡‡ˆϐ‹…‹‡…›‘ˆ–Š‡Ǧͳ•‹ƒ’’”‘ƒ…Šǡ™‡’”‡˜‹‘—•Ž› –‡•–‡†„›“Ǧ–Š‡ƒ‘—–‘ˆǦͳ‹”‡Žƒ–‹‘–‘…‘””‡•’‘†‡–Š Ž‡˜‡Ž•‹‡ƒ…‡ŽŽ•™Š‹…Š™‡”‡–”ƒ•ˆ‡…–‡†™‹–Šƒ–‹ǦŠ—ƒǦͳ•‹‹͵ƒ†‹”‘Ǧ –”‡ƒ–‡† …‡ŽŽ•Ǥ Ǧͳ  ™ƒ• ˆ‘—† –‘ „‡ †‡…”‡ƒ•‡† –‘ ‘”‡ –Šƒ ͷͲΨ ƒˆ–‡” Ǧͳ ‰‡‡ •‹Ž‡…‹‰‹͵Ǧƒ†‹”‘Ǧ–”‡ƒ–‡†…‡ŽŽ•ȋ ‹‰Ǥ͹Ȍȋ’δͲǤͲͷȌǤ

͵ ‹ˆ‡…–‹‘ –”‹‰‰‡”• ƒ…”‘’Šƒ‰‡• –‘ ‹ϐ‹Ž–”ƒ–‡ –Š‡ Š‡ƒ”–ǡ ”‡Ž‡ƒ•‹‰ ‹ ȏͷȐ and superoxide anions which are suggested to contribute to oxidative stress and cardiac damage ‹ …Š”‘‹… ˜‹”ƒŽ ›‘…ƒ”†‹–‹• ȏ͹ȐǤ ‡”‡ǡ ™‡ †‡‘•–”ƒ–‡ –Šƒ– ‹ –Š‡ …Š”‘‹… …‘—”•‡ ‘ˆ –Š‡ †‹•‡ƒ•‡ƒ…”‘’Šƒ‰‡•‘ˆȀ ‹…‡ǡ„—–‘––Š‘•‡‘ˆ”‡•‹•–ƒ–ͷ͹Ȁ͸‹…‡ǡ”‡˜‡ƒŽŠ‹‰Š ƒ‘—–•‘ˆ‹”‘ǡ™Š‹…Š‹•ƒŽ•‘‘™–‘—’”‡‰—Žƒ–‡’”‘†—…–‹‘ȏͳͺȐǤ‡…‡–Ž›ǡƒ•–”‘‰

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Discussion

Cellular Physiology and Biochemistry

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014



Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

control HeLa

A

© 2014 S. Karger AG, Basel www.karger.com/cpb

Fe

CVB3 + Fe

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D CVB3 + Fe

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CVB3 hHO-1siRNA

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Fe hHO-1 siRNA

Fig. 7.Ǧͳ•‹Ž‡…‹‰‹‡ƒ…‡ŽŽ•…‘””‡Žƒ–‡•™‹–Š”‡†—…–‹‘‘ˆ…ƒ•’ƒ•‡Ǧ͵ƒ…–‹˜ƒ–‹‘ǤǤ…–‹˜ƒ–‡†…ƒ•’ƒ•‡Ǧ͵ ’”‘–‡‹‡š’”‡••‹‘ȋ‰”‡‡Ȍ‹‡ƒ…‡ŽŽ•ƒˆ–‡”‹”‘ƒ†Ȁ‘”͵–”‡ƒ–‡–Ǥ͵ͳ’”‘–‡‹ȋ”‡†Ȍƒ•™‡ŽŽ ƒ•…ƒ•’ƒ•‡Ǧ͵ȋ‰”‡‡Ȍƒ”‡‡˜‡Ž›†‹•–”‹„—–‡†‹–Š‡…›–‘’Žƒ•‘ˆ–Š‡…‡ŽŽǤŠ‡•‹—Ž–ƒ‡‘—•’”‡•‡…‡‘ˆ͵ ƒ†‹”‘‹†—…‡•ƒ…–‹˜ƒ–‹‘‘ˆ…ƒ•’ƒ•‡Ǧ͵‹–Š‡—…Ž‡—•ƒ†…›–‘’Žƒ•ǤǤ‡…”‡ƒ•‡†…ƒ•’ƒ•‡Ǧ͵’”‘–‡‹‡š’”‡••‹‘‹…‡ŽŽ•‹•‘„•‡”˜‡†„›†‘™”‡‰—Žƒ–‹‘‘ˆǦͳ—•‹‰•‹‹ƒŽŽ–”‡ƒ–‡†•ƒ’Ž‡•ǤǤ‡’”‡•‡–ƒ–‹˜‡“—ƒ–‹ϐ‹…ƒ–‹‘‘ˆ…ƒ•’ƒ•‡Ǧ͵‡š’”‡••‹‘™‹–Šƒ†™‹–Š‘—–Ǧͳ•‹‹͵Ǧ‹ˆ‡…–‡†ƒ†‹”‘Ǧ–”‡ƒ–‡† …‡ŽŽ•Ǥ‹š‡Ž‹–‡•‹–‹‡•™Š‹…Š™‡”‡†‡–‡”‹‡†ˆ‘”‹†‹˜‹†—ƒŽ…‡ŽŽ•‘ˆ•‹‹Žƒ”•‹œ‡•ƒŽ‘‰ƒŽ‹‡…”‘••‹‰–Š‡ †‹ƒ‡–‡”‘ˆ–Š‡…‡ŽŽ‘—–•‹†‡‘ˆ–Š‡—…Ž‡—•”‡˜‡ƒŽƒ•‹‰‹ϐ‹…ƒ–†‘™”‡‰—Žƒ–‹‘‘ˆ…ƒ•’ƒ•‡Ǧ͵‹’”‡•‡…‡‘ˆ Ǧͳ•‹ȋȗȗȗ’δͲǤͲͲͳȌǤǤŠ‡‡ˆϐ‹…‹‡…›‘ˆ–Š‡Ǧͳ•‹ƒ’’”‘ƒ…Š™ƒ•ϐ‹”•––‡•–‡†„›“ǦǤǦͳ ™ƒ•ˆ‘—†–‘„‡†‡…”‡ƒ•‡†–‘‘”‡–ŠƒͷͲΨƒˆ–‡”Ǧͳ‰‡‡•‹Ž‡…‹‰‹͵ƒ†‹”‘Ǧ–”‡ƒ–‡†…‡ŽŽ• ȋ ‹‰Ǥ͹Ȍȋȗ’δͲǤͲͷȌǤ…ƒŽ‡„ƒ”ǣͳͲρǤ

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15

fold change of hHO-1 mRNA expression

CVB3 + Fe HO-1 siRNA

Signal intensity

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*

*

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Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb



Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

“—ƒ–‹–ƒ–‹˜‡ –”ƒ‹– Ž‘…—• ‹†‡–‹ϐ‹‡† ‘ ‘—•‡ …Š”‘‘•‘‡ ͻ ™ƒ• ’”‘˜‡ –‘ †‹ˆˆ‡”‡–‹ƒŽŽ› ƒˆˆ‡…––Š‡ƒ…”‘’Šƒ‰‡‹”‘„—”†‡‹Ȁ ƒ†ͷ͹•–”ƒ‹•ȏͳͻȐǡ…‘…Ž—†‹‰–Šƒ–‰‡‡–‹… „ƒ…‰”‘—†‹‰Š–‹ϐŽ—‡…‡–Š‡‹–”ƒ…‡ŽŽ—Žƒ”‹”‘‡–ƒ„‘Ž‹•‹–Š‡–™‘‘—•‡•–”ƒ‹•Ǥ  our manuscript, we provide new evidence that differences in macrophage iron content also ‹ϐŽ—‡…‡•–Š‡‘—–…‘‡‘ˆ–Š‡‡–‡”‘˜‹”ƒŽ›‘…ƒ”†‹–‹•‹–Š‡•‡‰‡‡–‹…ƒŽŽ›†‹ˆˆ‡”‡–‘—•‡ •–”ƒ‹•ǡ•—„•–ƒ–‹ƒ–‹‰–Š‡”‡Ž‡˜ƒ…‡‘ˆ–Š‡Š‘•–Ʋ‰‡‡–‹…•‹–Š‡’ƒ–Š‘‰‡‡•‹•‘ˆ–Š‡†‹•‡ƒ•‡Ǥ –‡”‡•–‹‰Ž›ǡ ‹…‘”’‘”ƒ–‹‘ ‘ˆ ‹”‘ ™ƒ• ‘– ‘Ž› ‘„•‡”˜‡† ‹ ƒ…”‘’Šƒ‰‡•ǡ „—– ƒŽ•‘ ‹ ͵Ǧ‹ˆ‡…–‡†…ƒ”†‹‘›‘…›–‡•‘ˆȀ ‹…‡™‹–Š‹ƒ”‡ƒ•‘ˆ‹…”‡ƒ•‡†‘š‹†ƒ–‹˜‡ƒ…–‹˜‹–‹‡•Ǥ ”‡˜‹‘—•Ž› ‹– ™ƒ• •Š‘™ ‹ ”ƒ– …ƒ”†‹‘›‘…›–‡• –Šƒ– ‹”‘ ‹†—…‡• ‡Šƒ…‡† ‰‡‡”ƒ–‹‘ ‘ˆ  ƒ† •–‹—Žƒ–‡• ‹ ƒ…–‹˜‹–›ǡ –Š—• …‘–”‹„—–‹‰ –‘ Š‡ƒ”– †ƒƒ‰‡ ȏʹͲȐǤ Š‡ ‡Šƒ…‡† …ƒ”†‹ƒ… ‘š‹†ƒ–‹˜‡ ƒ…–‹˜‹–‹‡• •‡‡ ‹ ͵Ǧ‹ˆ‡…–‡† •—•…‡’–‹„Ž‡ ‹…‡ ™‡”‡ …‘””‡Žƒ–‡† ™‹–Š –Š‡ —’”‡‰—Žƒ–‹‘ ‘ˆ –Š‡ Ǧͳ  ƒ† ’”‘–‡‹ ‹ ƒ…”‘’Šƒ‰‡•ǡ ƒ• ™‡ŽŽ ƒ• ‹ ‹ˆ‡…–‡† …ƒ”†‹‘›‘…›–‡•Ǥ‡‘ˆ–Š‡‘•–’‘–‡–‹†—…‡”•‘ˆǦͳ‰‡‡‡š’”‡••‹‘ˆ‘”˜ƒ”‹‘—•–‹••—‡• in vivoȏͳͷȐǡƒ•™‡ŽŽƒ•ˆ‘”†‹ˆˆ‡”‡–…‡ŽŽ–›’‡•in vitroȏͳ͸Ȑ ‹•Ǥ …”‡ƒ•‡†’”‘†—…–‹‘ ™ƒ•†‡•…”‹„‡†–‘…‘–”‹„—–‡–‘‹—‘’ƒ–Š‘Ž‘‰›‹‘‰‘‹‰—”‹‡͵›‘…ƒ”†‹–‹•ȏͷȐǤ  –Š‡ ‘–Š‡”Šƒ† ‹– ™ƒ• ’—„Ž‹•Š‡† –Šƒ– ‹ ’”‡•‡…‡‘ˆ ‡š…‡••‹˜‡ ǡ ’”‘†—…‡†„› ‡‹–Š‡” ‹‘˜‡”‡š’”‡••‹‘‘”–”‡ƒ–‡–™‹–Š‡š…‡••‹˜‡…‘…‡–”ƒ–‹‘•‘ˆƒ†‘‘”ǡ͵ replication is inhibited in vitroǦŠ‘™‡˜‡”‘Ž›†—”‹‰–Š‡‹‹–‹ƒŽ•–ƒ‰‡•‘ˆ‹ˆ‡…–‹‘™Š‡ ™ƒ•ƒ††‡†„‡ˆ‘”‡‘”Žƒ–‡•–ͳŠ‘—”’‘•–‹ˆ‡…–‹‘ȏʹͳȐǤ”‡˜‹‘—•Ž›‹–™ƒ••Š‘™–Šƒ–‹”ƒ–Ž‹˜‡” ‹”‘‘˜‡”Ž‘ƒ†–”‹‰‰‡”•‘š‹†ƒ–‹˜‡•–”‡••ƒ–‡ƒ”Ž›–‹‡•ǡ™‹–Š—’”‡‰—Žƒ–‹‘‘ˆ‹‡š’”‡••‹‘ ‹˜‘Ž˜‹‰–Š‡Ȁ Ǧƒ’’ƒ’ƒ–Š™ƒ›ƒ–Žƒ–‡”–‹‡•ȏʹʹȐǡƒ†–Šƒ–‹”ƒ–Š‡ƒ”–•‹†—…–‹‘ ‘ˆ…ƒ”†‹ƒ…‹’”‘‘–‡•›‘…ƒ”†‹ƒŽ‹Œ—”›†—‡–‘‘š‹†ƒ–‹˜‡•–”‡••ȏʹ͵Ȑ. —”‘„•‡”˜ƒ–‹‘•‹ •—•…‡’–‹„Ž‡Ȁ ‹…‡”‡˜‡ƒŽ‹‰…Š”‘‹…›‘…ƒ”†‹–‹•‹†‹…ƒ–‡–Šƒ––Š‡ȀǦͳ’ƒ–Š™ƒ› ‹•Ž‹‡Ž›‹†—…‡†„›„‘–Šǡ͵ƒ•™‡ŽŽƒ•‹”‘ǤIn vitro experiments revealed that blockage ‘ˆ’”‘†—…–‹‘„›Ǧ†‡…”‡ƒ•‡•Ǧͳ’”‘–‡‹‡š’”‡••‹‘‹͵ƒ†‹”‘Ǧ–”‡ƒ–‡† …‡ŽŽ•ǡ—†‡”Ž‹‹‰–Š‡”‘Ž‡‘ˆ‹‘†—Žƒ–‹‰Ǧͳƒ…–‹˜‹–›ǤŽ–Š‘—‰ŠƒŽ‘™‡š’”‡••‹‘Ž‡˜‡Ž ‘ˆǦͳ‹•†‡•…”‹„‡†–‘„‡ƒ••‘…‹ƒ–‡†™‹–Š’”‘–‡…–‹‘ƒ‰ƒ‹•–‘š‹†ƒ–‹˜‡•–”‡••ȏʹͶȐǡƒ‡š…‡•• expression of this enzyme was found to be associated with increased oxygen cytotoxicity, „‡‹‰ †‡–”‹‡–ƒŽ ˆ‘” –Š‡ …‡ŽŽ ’Š›•‹‘Ž‘‰› ȏʹͷȐǤ ‘–ƒ„Ž›ǡ ‹ ‘—” …‡ŽŽ …—Ž–—”‡ ‡š’‡”‹‡–• ™‡ ‘„•‡”˜‡† ‘– ‘Ž› ƒ …›–‘’Žƒ•‹…ǡ „—– ƒŽ•‘ ƒ ‹–”ƒ—…Ž‡ƒ” Ž‘…ƒŽ‹œƒ–‹‘ ‘ˆ Ǧͳ —†‡” ‹”‘ƒ†͵–”‡ƒ–‡–ǡ™Š‹…Š‹‰Š–„‡‡š’Žƒ‹‡†„›–Š‡ƒ––‡’–‘ˆ–Š‡…‡ŽŽ–‘’”‘˜‹†‡ cytoprotection against oxidative stress by regulating the expression of antioxidant genes ȏʹ͸ȐǤ Both superoxide and iron have been described to contribute to cardiac damage and …ƒ”†‹‘›‘…›–‡ƒ’‘’–‘•‹•ȏʹ͹ǡʹͺȐǤ‡‘„•‡”˜‡†ƒƒ…–‹˜ƒ–‹‘‘ˆ…ƒ•’ƒ•‡Ǧ͵in vitro as well as in vivo ‹…ƒ”†‹‘›‘…›–‡•‹ƒ••‘…‹ƒ–‹‘™‹–Š‹”‘‘˜‡”Ž‘ƒ†ƒ†͵‹ˆ‡…–‹‘Ǥ‘”‡˜‘˜‡”ǡ ™‡ˆ‘—†–Šƒ––Š‡•‡ƒ’‘’–‘–‹…’”‘…‡••‡•…‘—Ž†„‡‹Š‹„‹–‡†‹Ǧͳ•‹‡š’‡”‹‡–•Ǥ  ’”‡˜‹‘—•‡š’‡”‹‡–•™‹–ŠŽ‡—‡‹ƒ…‡ŽŽ•ǡ‹–™ƒ•ƒŽ•‘†‡‘•–”ƒ–‡†–Šƒ–Ǧͳ‡š’”‡••‹‘ …‘–”‹„—–‡•–‘ƒ’‘’–‘•‹•‡†‹ƒ–‡†„›‘š‹†ƒ–‹˜‡•–”‡••ȏʹͻȐǤ‘™‡˜‡”ǡ‹ƒ‘†‡Ž‘ˆ…ƒ”†‹ƒ… ‹•…Š‡‹ƒƒ†”‡’‡”ˆ—•‹‘ǡǦͳŠƒ•„‡‡†‡•…”‹„‡†–‘ƒ––‡—ƒ–‡…ƒ”†‹‘›‘…›–‹…ƒ’‘’–‘•‹• ȏ͵ͲȐ „› ”‡†—…‹‰  ǦN ƒ† Ǧͳ —…Ž‡ƒ” –”ƒ•Ž‘…ƒ–‹‘ǡ –Š‡”‡„› ”‡†—…‹‰ –Š‡ Ž‡˜‡Ž ‘ˆ …ƒ•’ƒ•‡Ǧ͵ƒ…–‹˜ƒ–‹‘ƒ†–Š‡•—”‰‡‘ˆ‹ϐŽƒƒ–‘”›…›–‘‹‡•ȏ͵ͳȐǤ–Š‡‘–Š‡”Šƒ†ǡ‘”‡ ”‡…‡–ϐ‹†‹‰••Š‘™‡†ƒ††‹–‹‘ƒŽ†‡Ž‡–‡”‹‘—•‡ˆˆ‡…–•‘ˆǦͳ„›’”‘˜‹‰–Šƒ–•›•–‡‹…Ǧͳ ‘˜‡”‡š’”‡••‹‘ƒ‰‰”ƒ˜ƒ–‡•’”‡••—”‡‘˜‡”Ž‘ƒ†Ǧ‹†—…‡†…ƒ”†‹ƒ…Š›’‡”–”‘’Š›ȏ͵ʹȐǤ ’‘”–ƒ–Ž›ǡ increased cellular iron deposition not only contributes to the heart disease by potentiating ‘š‹†ƒ–‹˜‡•–”‡••ƒ†ƒ’‘’–‘•‹•ǡ„—–ƒŽ•‘„›‹…”‡ƒ•‹‰˜‹”ƒŽ”‡’Ž‹…ƒ–‹‘™‹–ŠƒͶǦˆ‘Ž†‹…”‡ƒ•‡ ‹͵–”ƒ•…”‹’–—„‡”•‹‹”‘Ǧ–”‡ƒ–‡†‡ƒ…‡ŽŽ•Ǥ‘ˆƒ”ǡ–Š‡‘Ž‡…—Žƒ”„ƒ•‹•„›™Š‹…Š ‹”‘ ƒŽ–‡”• ͵ ”‡’Ž‹…ƒ–‹‘ ‹• —‘™Ǥ ‘™‡˜‡”ǡ ƒŽ•‘ ‹ ‘–Š‡” ˜‹”—• ‹ˆ‡…–‹‘•ǡ •—…Š ƒ•  Ǧͳǡ ƒ ‹…”‡ƒ•‡† ‹”‘ Ž‘ƒ† ‹ ƒ…”‘’Šƒ‰‡• ™ƒ• ˆ‘—† –‘ „‡ ƒ••‘…‹ƒ–‡† ™‹–Š ‹…”‡ƒ•‡† ˜‹”ƒŽ”‡’Ž‹…ƒ–‹‘ȏ͵͵ȐǤ‹‹Žƒ”Ž›ǡŠ‡’ƒ–‹–‹•˜‹”—•ƒ†‡š’”‡••‹‘™ƒ••Š‘™–‘ „‡‹…”‡ƒ•‡†„›‹”‘Ǥ –‡”‡•–‹‰Ž›ǡ‹‡š’”‡••‹‘•–—†‹‡•”‡˜‡ƒŽ‡†‘†—Žƒ–‘”›‡ˆˆ‡…–• ‘ˆ‹”‘‘”‡’Ž‹…ƒ–‹‘ȏ͵ͶȐǤŠ‡–Š‡”†‹ˆˆ‡”‡–‹ƒŽ‹‡š’”‡••‹‘‹‰Š–ƒŽ•‘’Žƒ›ƒ ”‘Ž‡ˆ‘”–Š‡‹…”‡ƒ•‡†͵”‡’Ž‹…ƒ–‹‘‹‘—”•—•…‡’–‹„Ž‡ƒ‹ƒŽ•Šƒ•–‘„‡‹˜‡•–‹‰ƒ–‡†‹ ˆ—”–Š‡”‡š’‡”‹‡–•Ǥ

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Cellular Physiology and Biochemistry

Cellular Physiology and Biochemistry

Cell Physiol Biochem 2014;33:52-66 DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb



Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

•—ƒ”›ǡ™‡Šƒ˜‡†‡‘•–”ƒ–‡†–Šƒ–Ǧͳ‡†‹ƒ–‡•ƒ’ƒ–Š‘Ž‘‰‹……”‘••–ƒŽ„‡–™‡‡ ƒ…”‘’Šƒ‰‡•ƒ†…ƒ”†‹‘›‘…›–‡•‹͵Ǧ‹†—…‡†›‘…ƒ”†‹–‹•ǡ–Š—•”‡•—Ž–‹‰‹‹…”‡ƒ•‡† ‘š‹†ƒ–‹˜‡ •–”‡•• ƒ† –Š‡ ‡˜‘Ž˜‡‡– ‘ˆ ƒ’‘’–‘–‹… …ƒ”†‹‘›‘…›–‡•ǡ ϐ‹ƒŽŽ› …‘–”‹„—–‹‰ –‘ Š‡ƒ”– ˆƒ‹Ž—”‡Ǥ ”‘ –Š‡•‡ †ƒ–ƒ ‹– …ƒ „‡ …‘…Ž—†‡† –Šƒ– ‹ ‰‡‡–‹…ƒŽŽ› •—•…‡’–‹„Ž‡ ‹…‡ ‹”‘•—„•–ƒ–‹ƒŽŽ›‹…”‡ƒ•‡•–Š‡†‡–”‹‡–ƒŽ‡ˆˆ‡…–•‘ˆ͵„›–Š‡ȀǦͳ’ƒ–Š™ƒ›ǡ–Š—• ‹…”‡ƒ•‹‰–Š‡’ƒ–Š‘‰‡‹…‹–›ƒ†‹Œ—”›‹–Š‡Š‡ƒ”–Ǥ•ƒ…‘•‡“—‡…‡ǡ’ƒ–‹‡–••—ˆˆ‡”‹‰ from chronic heart failure should not be prescribed iron unless it is proven that this severe †‹•‡ƒ•‡‹•‘––Š‡”‡•—Ž–‘ˆƒ…Š”‘‹…‡–‡”‘˜‹”—•‹ˆ‡…–‹‘Ǥ &RQÁLFWRI,QWHUHVW

‘‡Ǥ Acknowledgements

‡ –Šƒƒ†”ƒ—†•…Š—Šˆ‘”‡š…‡ŽŽ‡––‡…Š‹…ƒŽƒ••‹•–ƒ…‡ǤŠ‹•™‘”™ƒ••—’’‘”–‡† „›ƒ‰”ƒ–‘ˆ–Š‡ ȋ‘ǤͲͳͲͺͳ͹Ȍƒ†–Š‡‡—–•…Š‡ ‘”•…Š—‰•‰‡‡‹•…Šƒˆ–ȋ Ǧ ͳͻȌ–‘ǤǤƒ†ǤǤ

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ƒ”–‹‘ǡ‹—ǡ‘Ž‡ ǣ‹”ƒŽ‹ˆ‡…–‹‘ƒ†–Š‡’ƒ–Š‘‰‡‡•‹•‘ˆ†‹Žƒ–‡†…ƒ”†‹‘›‘’ƒ–Š›Ǥ‹”…‡• ͳͻͻͶǢ͹ͶǣͳͺʹǦͳͺͺǤ ƒƒ•

ǣ–‹‘Ž‘‰›ǡ‡˜ƒŽ—ƒ–‹‘ǡƒ†ƒƒ‰‡‡–‘ˆƒ…—–‡›‘…ƒ”†‹–‹•Ǥƒ”†‹‘Ž‡˜ʹͲͲͳǢͻǣͺͺǦͻͷǤ …ƒ—•ǡŠ‘™ǡ‹Ž•‘ ǡ†‡”•‘ǡ —Ž‹œ‹ƒ ǡ ƒ—–– ǡŽ‹‰‡Žǡ‡‹•‡Žǡƒ†‘Žˆ ǣ‹”‡…–›‘…ƒ”†‹ƒŽ‹Œ—”›„›‡–‡”‘˜‹”—•ǣƒ…‡–”ƒŽ”‘Ž‡‹–Š‡‡˜‘Ž—–‹‘‘ˆ—”‹‡›‘…ƒ”†‹–‹•ǤŽ‹ —‘Ž —‘’ƒ–Š‘Žͳͻͻ͵Ǣ͸ͺǣͳͷͻǦͳ͸ͻǤ ‹ǡ—ǡ —‘ǡ ‹ƒ‰ǡƒ‰ǡ—‡ǡ‹‘‰ǣ‹ˆˆ‡”‡–‹ƒŽƒ…”‘’Šƒ‰‡’‘Žƒ”‹œƒ–‹‘‹ƒŽ‡ƒ† ˆ‡ƒŽ‡Ȁ…‹…‡‹ˆ‡…–‡†™‹–Š…‘š•ƒ…‹‡˜‹”—•͵†‡ϐ‹‡••—•…‡’–‹„‹Ž‹–›–‘˜‹”ƒŽ›‘…ƒ”†‹–‹•Ǥ‹”…‡• ʹͲͲͻǢͳͲͷǣ͵ͷ͵Ǧ͵͸ͶǤ œƒŽƒ› ǡƒ—–‡”ǡƒŽ† ǡ‡‹œ‹‡”Žǡƒ†‘ŽˆǡŽ‹‰‡Žǣ—•–ƒ‹‡†‹–”‹…‘š‹†‡•›–Š‡•‹•…‘–”‹„—–‡• –‘‹—‘’ƒ–Š‘Ž‘‰›‹‘‰‘‹‰›‘…ƒ”†‹–‹•ƒ––”‹„—–ƒ„Ž‡–‘ –‡”Ž‡—‹ǦͳͲ†‹•‘”†‡”•Ǥ ƒ–Š‘Ž ʹͲͲ͸Ǣͳ͸ͻǣʹͲͺͷǦʹͲͻ͵Ǥ Šƒ†‘—” ǡƒƒ•ǡ ‡”†‹ƒ†›ǡ…Š—Žœ‡ǡ•‘–ǡƒŽ—ǡ‹Ž†Š‹”–ǡ…Š—ŽœǣŠƒ…‡† ƒ†•—’‡”‘š‹†‡‰‡‡”ƒ–‹‘‹†›•ˆ—…–‹‘ƒŽŠ‡ƒ”–•ˆ”‘‡†‘–‘š‡‹…”ƒ–•Ǥ Š›•‹‘Ž‡ƒ”–‹”…Š›•‹‘Ž ʹͲͲʹǢʹͺ͵ǣͳͳͲͺǦͳͳͳͷǤ ‹”ƒ‘ƒǡ‹•Š‹‘–‘ǡƒƒ†ƒǡ—”‘ƒ™ƒǡ…Š‹ƒ‹ǡŠ‹”ƒ‹ǡƒ•ƒ›ƒƒǣ‘Ž‡‘ˆ‘š›‰‡†‡”‹˜‡† ˆ”‡‡”ƒ†‹…ƒŽ•‹–Š‡’ƒ–Š‘‰‡‡•‹•‘ˆ…‘š•ƒ…‹‡˜‹”—•͵›‘…ƒ”†‹–‹•‹‹…‡Ǥƒ”†‹‘˜ƒ•…‡•ͳͻͻ͵Ǣʹ͹ǣͻͷ͹Ǧ ͻ͸ͳǤ ›–‡”ǡ––‡”„‡‹ǡ‘”•‡ǡŠ‘‹ǣ‡‡‘š›‰‡ƒ•‡Ȁ…ƒ”„‘‘‘š‹†‡•‹‰ƒŽ‹‰’ƒ–Š™ƒ›•ǣ ”‡‰—Žƒ–‹‘ƒ†ˆ—…–‹‘ƒŽ•‹‰‹ϐ‹…ƒ…‡Ǥ‘Ž‡ŽŽ‹‘…Š‡ʹͲͲʹǢʹ͵ͶǦʹ͵ͷǣʹͶͻǦʹ͸͵Ǥ ‹Ž‡ ǡƒ•—Ǧ‘†ƒǡƒŽ–‡”ǡ›””‡ŽŽǣ‡‡‘š›‰‡ƒ•‡ͳ‡†‹ƒ–‡•ƒƒ†ƒ’–‹˜‡”‡•’‘•‡–‘ ‘š‹†ƒ–‹˜‡•–”‡••‹Š—ƒ•‹ϐ‹„”‘„Žƒ•–•Ǥ”‘…ƒ–Ž…ƒ†…‹ͳͻͻͶǢͻͳǣʹ͸Ͳ͹Ǧʹ͸ͳͲǤ ‹‰ǡŠ‡ǡƒ„ ǡ—„›ǡ—‹Žƒ

ǡ˜ƒ•ǡ —––‡”‹†‰‡ ǣ ”‘‘˜‡”Ž‘ƒ†—’”‡‰—Žƒ–‡• Šƒ‡‘š›‰‡ƒ•‡ͳ‹–Š‡Ž—‰‘”‡”ƒ’‹†Ž›–Šƒ‹‘–Š‡”–‹••—‡•Ǥ ‡––ͳͻͻͻǢͶͶ͹ǣͳͳͳǦͳͳͶǤ

ƒ‘™•ƒǡ‘œ‡–”›–ǡ‹–‘™•ƒǡ‘™ƒ ǡƒ”–ƒǡ‘‹‘™•ƒǡ‘”‘†—Ž‹Ǧƒ†œ‹‡Œƒǡ ƒƒ•‹ƒǡ‘Ž‘•‹ǡ ‹Ž‹’’ƒ–‘• ǡ…—””ƒ›

ǡ‡”ǡ‘‹‘™•‹ǣ ”‘†‡ϐ‹…‹‡…›ǣƒ‘‹‘—• •‹‰‹’ƒ–‹‡–•™‹–Š•›•–‘Ž‹……Š”‘‹…Š‡ƒ”–ˆƒ‹Ž—”‡Ǥ—”‡ƒ”– ʹͲͳͲǢ͵ͳǣͳͺ͹ʹǦͳͺͺͲǤ …Š™ƒ”–œǡ‹ǡ…Š™ƒ”–œǡ‘‘’‡” ǡ”ƒ•‡Ž–‘ǣƒ”Ž‹‡•–…ƒ”†‹ƒ…–‘š‹…‹–›‹†—…‡†„›‹”‘ ‘˜‡”Ž‘ƒ†•‡Ž‡…–‹˜‡Ž›‹Š‹„‹–•‡Ž‡…–”‹…ƒŽ…‘†—…–‹‘Ǥ ’’ŽŠ›•‹‘ŽʹͲͲʹǢͻ͵ǣ͹Ͷ͸Ǧ͹ͷͳǤ ‹ ǡ‘–Šƒ”…Š™‹„‡…ǡ•…Š‘’‡ǡ‡•…Šƒǡ—•ƒǡ—–•…Š‘™ǡ‡‹…Š‡„ƒ…Š ǡ‘—–•‹ƒ•ǡ ‘„ƒŽœǡ‘ŽŽ‡”ǡ’‹ŽŽƒ ǡ‡‹…ŠŠƒ”†–ǡ…Š—Ž–Š‡‹••ǡƒ—•…Š‹‰‡”ǣ‘ŽŽƒ‰‡†‡‰”ƒ†ƒ–‹‘‹ ƒ—”‹‡›‘…ƒ”†‹–‹•‘†‡Žǣ”‡Ž‡˜ƒ…‡‘ˆƒ–”‹š‡–ƒŽŽ‘’”‘–‡‹ƒ•‡‹ƒ••‘…‹ƒ–‹‘™‹–Š‹ϐŽƒƒ–‘”› ‹†—…–‹‘Ǥƒ”†‹‘˜ƒ•…‡•ʹͲͲʹǢͷ͸ǣʹ͵ͷǦʹͶ͹Ǥ

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Cellular Physiology and Biochemistry

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DOI: 10.1159/000356649 Published online: January 02, 2014

© 2014 S. Karger AG, Basel www.karger.com/cpb



Ursu et al.: Heme Oxygenase-1 in Enteroviral Myocarditis

Ž‹‰‡Žǡ‘Š‡ƒ†Žǡƒ—ǡŽ„”‡…Š–ǡ‡‡ƒǡƒŽŽ ǡƒ†‘Žˆǣ‰‘‹‰‡–‡”‘˜‹”—•Ǧ‹†—…‡† ›‘…ƒ”†‹–‹•‹•ƒ••‘…‹ƒ–‡†™‹–Š’‡”•‹•–‡–Š‡ƒ”–—•…Ž‡‹ˆ‡…–‹‘ǣ“—ƒ–‹–ƒ–‹˜‡ƒƒŽ›•‹•‘ˆ˜‹”—•”‡’Ž‹…ƒ–‹‘ǡ –‹••—‡†ƒƒ‰‡ǡƒ†‹ϐŽƒƒ–‹‘Ǥ”‘…ƒ–Ž…ƒ†…‹ͳͻͻʹǢͺͻǣ͵ͳͶǦ͵ͳͺǤ ‹ŽŽ‹•ǡ‘Ž‹•‘ǡ ”‡†‡”‹…ǡƒ—Ž…Žƒ” ǡ‹ŽŽ‘—‰Š„›ǣ‘†—Žƒ–‹‘‘ˆŠ‡‡‘š›‰‡ƒ•‡ ƒ…–‹˜‹–›‹”ƒ–„”ƒ‹ƒ†•’Ž‡‡„›‹Š‹„‹–‘”•ƒ††‘‘”•‘ˆ‹–”‹…‘š‹†‡Ǥ‹‘…Š‡‹‘’Š›•‡•‘— ͳͻͻͷǢʹͳͶǣͳͳͷʹǦͳͳͷ͸Ǥ ƒ––ƒǡ‹ƒ‘•ǣ‹–”‹…‘š‹†‡‹†—…‡•Š‡‡‘š›‰‡ƒ•‡Ǧͳ‰‡‡‡š’”‡••‹‘‹‡•ƒ‰‹ƒŽ…‡ŽŽ•Ǥ‹†‡› – ͳͻͻͻǢͷͷǣͳ͹͵ͶǦͳ͹͵ͻǤ ‹ ǡ ‡–‹Ž‡ǡ„”ƒŠƒ ǡƒ‡ŽŽƒǡƒƒ˜‘ ǡƒœœƒ ǡ—””‘ǡƒ…‹–‹ ǡ˜‘Žƒǡƒ’‹•‹ǣ —‘…›–‘…Š‡‹…ƒŽŽ‘…ƒŽ‹œƒ–‹‘ƒ†‡š’”‡••‹‘‘ˆŠ‡‡‘š›‰‡ƒ•‡Ǧͳ‹’”‹ƒ”›ƒ•–”‘‰Ž‹ƒŽ…‡ŽŽ…—Ž–—”‡• †—”‹‰†‹ˆˆ‡”‡–‹ƒ–‹‘ǣ‡ˆˆ‡…–‘ˆ‰Ž—–ƒƒ–‡Ǥ‹‘…Š‡‹‘’Š›•‡•‘—ʹͲͲͶǢ͵ͳͷǣͷͳ͹ǦͷʹͶǤ ›ƒǡ—•–ǣŠ‡”‘Ž‡‘ˆ‹”‘‹‘š›‰‡Ǧ‡†‹ƒ–‡†–‘š‹…‹–‹‡•Ǥ”‹–‡˜‘š‹…‘ŽͳͻͻʹǢʹʹǣͳͳͻǦͳͶͳǤ ƒ‰ ǡƒ”ƒ†ƒ”ǡ—•–‘†‹‘ǡ‡›ƒ”†ǡ Ž‡‹‰ǡƒ’ƒ‰ƒǡ‘„‡”–•ǡ‘›ƒ”–…Š—ǡ ‹‡–”‹…Š ǡƒ’Žƒ ǡ†”‡™•ǣ ‡‡–‹…˜ƒ”‹ƒ–‹‘‹‘ͳƒƒˆˆ‡…–•’”‘–‡‹–”ƒˆϐ‹…‹‰ƒ†‘†‹ϐ‹‡• ƒ…”‘’Šƒ‰‡‹”‘Ž‘ƒ†‹‰‹‹…‡Ǥƒ– ‡‡–ʹͲͲ͹Ǣ͵ͻǣͳͲʹͷǦͳͲ͵ʹǤ —‘œ ǡŠ‹‘‰ǡ ƒ”…Àƒǡ”‘…‘•‘ǡ‘”‘ǡ‡†”‘œ‘ǡ‹ƒœǦŽ‹œ‘†‘ ǡƒŽƒ•ǡƒ””ƒǡïӇœǡ ‹†ƒŽ‰‘ǡƒ˜ƒ†‡”‘ǣ ”‘‹†—…‡•’”‘–‡…–‹‘ƒ†‡…”‘•‹•‹…—Ž–—”‡†…ƒ”†‹‘›‘…›–‡•ǣ”‘Ž‡‘ˆ”‡ƒ…–‹˜‡ ‘š›‰‡•’‡…‹‡•ƒ†‹–”‹…‘š‹†‡Ǥ ”‡‡ƒ†‹…‹‘Ž‡†ʹͲͳͲǢͶͺǣͷʹ͸Ǧͷ͵ͶǤ ƒ”ƒ‰‘œƒǡ…ƒ’‘ ǡƒ—”ƒǡ…‹ŽŽƒǡ‘™‡•–‡‹ ǣ‹–”‹…‘š‹†‡‹Š‹„‹–‹‘‘ˆ…‘š•ƒ…‹‡˜‹”—• ”‡’Ž‹…ƒ–‹‘‹˜‹–”‘Ǥ Ž‹ ˜‡•–ͳͻͻ͹ǢͳͲͲǣͳ͹͸ͲǦͳ͹͸͹Ǥ ‘”‡Œ‘ǡƒ”‡Žƒǡ‹†‡Žƒǡ ‡”ž†‡œǣŠ”‘‹…‹”‘‘˜‡”Ž‘ƒ†‡Šƒ…‡•‹†—…‹„Ž‡‹–”‹…‘š‹†‡ •›–Šƒ•‡‡š’”‡••‹‘‹”ƒ–Ž‹˜‡”Ǥ‹–”‹…š‹†‡ʹͲͲͷǢͳ͵ǣͷͶǦ͸ͳǤ ‰ƒ”ƒ•Š‹ ǡ‹•Š‹†ƒǡ‘•Š‹†ƒǡƒƒ•Š‹–ƒǡ‘•ƒƒǡ‘”‹ǡ—œ—›ƒǡƒ†ƒǣ †—…‹„Ž‡‹–”‹…‘š‹†‡ •›–Šƒ•‡ƒ—‰‡–•‹Œ—”›‡Ž‹…‹–‡†„›‘š‹†ƒ–‹˜‡•–”‡••‹”ƒ–…ƒ”†‹ƒ…›‘…›–‡•Ǥ Š›•‹‘Ž‡ŽŽŠ›•‹‘Ž ͳͻͻͺǢʹ͹ͶǣʹͶͷǦʹͷʹǤ ‡‡”›ǡ‘‰ǡ”‹†Šƒ” ǡ‘†‰‡”•ǡ‹ ǡ’‹–œǣ‹ˆˆ‡”‡…‡•‹„ƒ•ƒŽƒ†Š›’‡”‘š‹ƒǦ ƒ••‘…‹ƒ–‡†‡š’”‡••‹‘‹‘š‹†ƒ–Ǧ”‡•‹•–ƒ–Šƒ•–‡”ϐ‹„”‘„Žƒ•–•Ǥ Š›•‹‘Ž—‰‡ŽŽ‘ŽŠ›•‹‘Ž ͳͻͻ͸Ǣʹ͹ͳǣ͸͹ʹǦ͸͹ͻǤ —––‡”ǡ‡‡”›ǣ‡˜‡”•ƒŽ‘ˆǦͳ”‡Žƒ–‡†…›–‘’”‘–‡…–‹‘™‹–Š‹…”‡ƒ•‡†‡š’”‡••‹‘‹•†—‡–‘ ”‡ƒ…–‹˜‡‹”‘Ǥ  ͳͻͻͻǢͳ͵ǣͳͺͲͲǦͳͺͲͻǤ ‹ǡ‡‹•ǡƒ‰ ǡ‡‰ǡ‡Ž•–‘ǡ‹•Šǡ‹–Šǡ‘”†‡” ǡ‘Ž–‡ǡ ƒ—‹–œ ǡ‡‡”›ǣ ‡‡‘š›‰‡ƒ•‡Ǧͳ’”‘–‡‹Ž‘…ƒŽ‹œ‡•–‘–Š‡—…Ž‡—•ƒ†ƒ…–‹˜ƒ–‡•–”ƒ•…”‹’–‹‘ˆƒ…–‘”•‹’‘”–ƒ–‹ ‘š‹†ƒ–‹˜‡•–”‡••Ǥ ‹‘ŽŠ‡ʹͲͲ͹ǢʹͺʹǣʹͲ͸ʹͳǦʹͲ͸͵͵Ǥ ‹ ǡ‹‡–œǡ˜‘ƒ”•†‘”ˆǣ—’‡”‘š‹†‡‹†—…‡•ƒ’‘’–‘•‹•‹…ƒ”†‹‘›‘…›–‡•ǡ„—–’”‘Ž‹ˆ‡”ƒ–‹‘ƒ† expression of transforming growth factor-Eͳ‹…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ ‡––ͳͻͻͻǢͶͶͺǣʹͲ͸ǦʹͳͲǤ ƒ‰ǡ—ǡŽǦ‘—•ƒǡ‹—ǡ ƒ‹ ǡƒ–—”‹ǡ”˜ƒ’ƒŽŽ‹ǡƒ––ƒǡƒƒ”Žƒǡ‹…‡ǡ”‹‡•– ǡŽ‘—‰Šǣ ”‘Ǧ‹†—…‡†…ƒ”†‹ƒ…†ƒƒ‰‡ǣ”‘Ž‡‘ˆƒ’‘’–‘•‹•ƒ††‡ˆ‡”ƒ•‹”‘š‹–‡”˜‡–‹‘Ǥ Šƒ”ƒ…‘Ž š’Š‡”ʹͲͳͳǢ͵͵͸ǣͷ͸Ǧ͸͵Ǥ ‹‹–‘‡ǡ‹‹–‘‡ǡ‹—Žƒǡƒ–›ƒƒǡƒ˜‘Žƒ‹‡ǡ‘‹•–‹‡ǣ‘–”‹„—–‹‘‘ˆ–Š‡‹†—…–‹‘ ‘ˆŠ‡‡‘š›‰‡ƒ•‡Ǧͳ–‘‡–‘’‘•‹†‡Ǧ‹†—…‡†ƒ’‘’–‘•‹•‹ƒ…—–‡›‡Ž‘„Žƒ•–‹…Ž‡—‡‹ƒǤƒ‡ƒ–‘Ž‘‰‹…ƒ ʹͲͲ͵ǢͺͺǣͳͳʹǦͳͳ͵Ǥ

‹ƒ‰ǡŠ‡ǡƒ‰ǡƒ ǡŠ‡ǡ‹ǡŠ—ǡƒ‘ǡ‹—ǣǦͳ‰‡‡‘˜‡”‡š’”‡••‹‘‡Šƒ…‡•–Š‡ „‡‡ϐ‹…‹ƒŽ‡ˆˆ‡…–•‘ˆ•—’‡”’ƒ”ƒƒ‰‡–‹…‹”‘‘š‹†‡Žƒ„‡Ž‡†„‘‡ƒ””‘™•–”‘ƒŽ…‡ŽŽ•–”ƒ•’Žƒ–ƒ–‹‘‹ •™‹‡Š‡ƒ”–•—†‡”™‡–‹•…Š‡‹ƒȀ”‡’‡”ˆ—•‹‘ǣƒ •–—†›Ǥƒ•‹…‡•ƒ”†‹‘ŽʹͲͳͲǢͳͲͷǣͶ͵ͳǦͶͶʹǤ ‡ŠǡŠ‡ǡƒ‰ǡ‹ǡ‹ ǣǦͳƒ…–‹˜ƒ–‹‘…ƒƒ––‡—ƒ–‡…ƒ”†‹‘›‘…›–‹…ƒ’‘’–‘•‹•˜‹ƒ inhibition of NF-Nƒ†Ǧͳ–”ƒ•Ž‘…ƒ–‹‘ˆ‘ŽŽ‘™‹‰…ƒ”†‹ƒ…‰Ž‘„ƒŽ‹•…Š‡‹ƒƒ†”‡’‡”ˆ—•‹‘Ǥ —”‰‡• ʹͲͲͻǢͳͷͷǣͳͶ͹Ǧͳͷ͸Ǥ Š‡ǡ—‘ǡ‘‰ǡŠ‡‰ǡ‹—ǡ ƒ‘ǡƒƒŒ‹ƒǡƒ‰ ǡ‘‰ǣ›•–‡‹…Š‡‡‘š›‰‡ƒ•‡Ǧͳ –”ƒ•‰‡‹…‘˜‡”‡š’”‡••‹‘ƒ‰‰”ƒ˜ƒ–‡•’”‡••—”‡‘˜‡”Ž‘ƒ†Ǧ‹†—…‡†…ƒ”†‹ƒ…Š›’‡”–”‘’Š›‹‹…‡Ǥ‡ŽŽ Š›•‹‘Ž‹‘…Š‡ʹͲͳͳǢʹͺǣʹͷǦ͵ʹǤ ”ƒ‘”‡ǡ‡›‡”ǣŠ‡‡ˆˆ‡…–‘ˆ‹”‘‘˜‡”Ž‘ƒ†‘‹˜‹–”‘ Ǧͳ‹ˆ‡…–‹‘Ǥ Ž‹‹”‘ŽʹͲͲͶǢ͵ͳǣͻʹǦͻͺǤ ƒ”ǡ—ƒ”ǡ —’–ƒǣ Ǧ„‡–ƒƒ†‹”‘†‹ˆˆ‡”‡–Ž›ƒŽ–‡””‡’Ž‹…ƒ–‹‘‹Š—ƒŠ‡’ƒ–‘…›–‡• –Š”‘—‰Š ǦEȀ•‹‰ƒŽ‹‰ƒ†…‡ŽŽ—Žƒ”‹…”‘‡š’”‡••‹‘Ǥ‘‡ʹͲͳʹǢ͹ǣ‡͵ͻʹ͹͸Ǥ

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Cell Physiol Biochem 2014;33:52-66