calcium sensing receptor promotes cardiac fibroblast proliferation and extracellular matrix secretion

Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating and Secretion Accepted: January 13, 2014 Cardiac Fibroblast Proliferation 1421-9778/14/0333-0557$39.50/0 This is an Open Access article licensed under the terms of the Creative Commons AttributionNonCommercial 3.0 Unported license (CC BY-NC) (www.karger.com/OA-license), applicable to the online version of the article only Distribution permitted for non-commercial purposes only Original Paper Calcium Sensing Receptor Promotes Cardiac Fibroblast Proliferation and Extracellular Matrix Secretion Xinying Zhanga,e Tiemin Zhangb,e Jichao Wua Xiangjing Yuc Dan Zhenga Fan Yanga Ting Li a Lina Wanga Yajun Zhaoa Shiyun Donga Xin Zhonga Songbin Fud Chang-qing Xua Fanghao Lua Wei-hua Zhanga Department of Pathophysioloy, dDepartment of Genetics, Harbin Medical University, bDepartment of *HQHUDO6XUJHU\WKH)LUVW$IÀOLDWHG+RVSLWDORI+DUELQ0HGLFDO8QLYHUVLW\ cClinical Laboratory, Harbin Cancer Hospital of Harbin Medical University, Harbin, 150086, China; erepresents equal contribution to this paper a Key Words &DOFLXPVHQVLQJUHFHSWRU‡&DUGLDFÀEUREODVW‡,QWUDFHOOXODUFDOFLXP‡&DUGLDFÀEURVLV Abstract Aims: Calcium-sensing receptor (CaR) acts as a G protein coupled receptor that mediates the increase of the intracellular Ca2+ FRQFHQWUDWLRQ 7KH H[SUHVVLRQ RI &D5 KDV EHHQ FRQÀUPHG in various cell types, including cardiomyocytes, smooth muscle cells, neurons and vascular HQGRWKHOLDOFHOOV+RZHYHUZKHWKHU&D5LVH[SUHVVHGDQGIXQFWLRQVLQFDUGLDFÀEUREODVWVKDV remained unknown The present study investigated whether CaR played a role in cardiac ÀEUREODVWSUROLIHUDWLRQDQGH[WUDFHOOXODUPDWUL[(&0VHFUHWLRQERWKLQFXOWXUHGUDWQHRQDWDO FDUGLDF ÀEUREODVWV DQG LQ D PRGHO RI FDUGLDF K\SHUWURSK\ LQGXFHG E\ LVRSURWHUHQRO ,62 Methods and Results:,PPXQRÁXRUHVFHQFHLPPXQRKLVWRFKHPLVWU\DQG:HVWHUQEORWDQDO\VLV UHYHDOHGWKHSUHVHQFHRI&D5LQFDUGLDFÀEUREODVWV&DOFLXPDQGFDOLQGRODVSHFLÀFDFWLYDWRU RI&D5HOHYDWHGWKHLQWUDFHOOXODUFDOFLXPFRQFHQWUDWLRQLQFDUGLDFÀEUREODVWV3UHWUHDWPHQWRI FDUGLDFÀEUREODVWVZLWKFDOKH[DVSHFLÀFLQKLELWRURI&D58DQG$3%DWWHQXDWHG the calindol- and extracellular calcium-induced increase in intracellular calcium ([Ca2+]i) &DUGLDFÀEUREODVWSUROLIHUDWLRQDQGPLJUDWLRQZHUHDVVHVVHGE\077'LPHWK\OWKLD]RO 2-yl)-2,5-diphenyltetrazolium bromide), cell count and the cell scratch assay ECM production was detected by expression of matrix metalloproteinase-3 and -9 (MMP-3 and -9) Activation RI &D5 SURPRWHG FDUGLDF ÀEUREODVW SUROLIHUDWLRQ DQG PLJUDWLRQ DQG (&0 VHFUHWLRQ 0RUH LPSRUWDQWO\FDOKH[VXSSUHVVHGFDUGLDFÀEUREODVWSUROLIHUDWLRQDQGPLJUDWLRQDQG003 DQGH[SUHVVLRQ7RIXUWKHULQYHVWLJDWHWKHHIIHFWRI&D5RQFDUGLDFÀEURVLVDPRGHORI ,62LQGXFHG FDUGLDF K\SHUWURSK\ ZDV HVWDEOLVKHG 3UHWUHDWPHQW ZLWK FDOKH[ SUHYHQWHG FDUGLDFÀEURVLVDQGGHFUHDVHGWKHH[SUHVVLRQRI003DQGH[SUHVVLRQConclusions: Our UHVXOWVDUHWKHÀUVWUHSRUWWKDW&D5SOD\VDQLPSRUWDQWUROHLQ&D2+ signaling involved in cardiac ÀEURVLVWKURXJKWKHSKRVSKROLSDVH&LQRVLWROSKRVSKDWH3/&,33) pathway Copyright © 2014 S Karger AG, Basel Wei-hua Zhang and Fanghao Lu Department of Pathophysioloy, Harbin Medical University, Harbin, 150086 (China) Fax +8645186674548, E-Mail zhangwh116@126.com and E-Mail lufanghao1973@126.com 557 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion Introduction ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•ƒ”‡–Š‡ƒ‹‘Ǧ›‘…›–‡…‡ŽŽ–›’‡‹–Š‡Š‡ƒ”–Ǥ†‡”’Š›•‹‘Ž‘‰‹…ƒŽ …‹”…—•–ƒ…‡•ǡ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• ’Žƒ› ƒ ‹’‘”–ƒ– ”‘Ž‡ ‹ ƒ‹–ƒ‹‹‰ Š‘‡‘•–ƒ•‹• ‘ˆ –Š‡ ‡š–”ƒ…‡ŽŽ—Žƒ” ƒ–”‹šǡ ‹ ƒ††‹–‹‘ –‘ ’”‘†—…‹‰ ‰”‘™–Š ˆƒ…–‘”•ǡ …›–‘‹‡• ƒ† ƒ–”‹š ‡–ƒŽŽ‘’”‘–‡‹ƒ•‡•ȋ•ȌȏͳȐǤ‘™‡˜‡”ǡ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•’”‘‘–‡›‘…ƒ”†‹ƒŽ•…ƒ””‹‰ ƒ†”‡‘†‡Ž‹‰‹‹Œ—”‡†Š‡ƒ”–•‹‹•…Š‡‹…ǡŠ›’‡”–‡•‹˜‡ƒ†Š›’‡”–”‘’Š‹…Š‡ƒ”–†‹•‡ƒ•‡•Ǥ š…‡••‹˜‡’”‘Ž‹ˆ‡”ƒ–‹‘ƒ†‹‰”ƒ–‹‘‘ˆ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•”‡•—Ž–‹ϐ‹„”‘•‹•ȏʹǡ͵ȐǤ›‘…ƒ”†‹ƒŽ ϐ‹„”‘•‹• …ƒ ƒŽ–‡” –Š‡ ‡…Šƒ‹…ƒŽ ’”‘’‡”–‹‡• ‘ˆ –Š‡ Š‡ƒ”–•ǡ ‹’ƒ‹”‹‰ …ƒ”†‹ƒ… ˆ—…–‹‘ ƒ† ’”‘‘–‹‰ Š‡ƒ”– ˆƒ‹Ž—”‡ ȏͶȐǤ ‘‡ ‡˜‹†‡…‡ Šƒ• •Š‘™ –Šƒ– –Š‡ …›–‘•‘Ž‹… …ƒŽ…‹— ‹‘ ȋȏƒʹΪȐiȌƒ…–•ƒ•ƒ•‡…‘†‡••‡‰‡”–‘”‡‰—Žƒ–‡˜‡”•ƒ–‹Ž‡…‡ŽŽˆ—…–‹‘•‹…Ž—†‹‰‡š…‹–ƒ–‹‘Ǧ …‘†‹–‹‘…‘—’Ž‹‰ǡ‰‡‡–”ƒ•…”‹’–‹‘ǡ…‡ŽŽ‰”‘™–Šǡ†‹ˆˆ‡”‡–‹ƒ–‹‘ƒ†ƒ’‘’–‘•‹•Ǥ‘™‡˜‡”ǡ Ž‹––Ž‡ ‹ˆ‘”ƒ–‹‘ ‹’Ž‹‡• –Šƒ– –Š‡ ƒʹΪ •‹‰ƒŽ‹‰ ’ƒ–Š™ƒ› ‹• ‹˜‘Ž˜‡† ‹ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•– ’”‘Ž‹ˆ‡”ƒ–‹‘ƒ†‡š–”ƒ…‡ŽŽ—Žƒ”ƒ–”‹šȋȌ’”‘†—…–‹‘ȏͷȐǤ ƒŽ…‹—•‡•‹‰”‡…‡’–‘”ȋƒȌǡƒ ’”‘–‡‹…‘—’Ž‡†”‡…‡’–‘”ǡƒ…–‹˜ƒ–‡•–Š‡’Š‘•’Š‘Ž‹’ƒ•‡ ȋȌ•‹‰ƒŽ‹‰’ƒ–Š™ƒ›ǡ”‡•—Ž–‹‰‹ƒ‹…”‡ƒ•‡‹‹‘•‹–‘Ž’Š‘•’Šƒ–‡ȋȌƒ……——Žƒ–‹‘ ƒ†ƒʹڏ‘„‹Ž‹œƒ–‹‘ȏ͸ȐǤ—”’”‡˜‹‘—•™‘”†‡‘•–”ƒ–‡†–Šƒ–ƒ‹†—…‡†…ƒ”†‹‘›‘…›–‡ ƒ’‘’–‘•‹•–Š”‘—‰Š–Š‡‹–‘…Š‘†”‹ƒŽƒ†‡†‘ȋ•ƒ”…‘Ȍ’Žƒ•‹…”‡–‹…—Ž—ƒ’‘’–‘–‹…’ƒ–Š™ƒ› ‹ –Š‡ ˆƒ‹Ž‹‰ Š‡ƒ”–Ǥ ‘”‡‘˜‡”ǡ ‘—” ”‡•—Ž–• •Š‘™‡† ‘„˜‹‘—• ›‘…ƒ”†‹ƒŽ ϐ‹„”‘•‹• ˆ‘ŽŽ‘™‹‰ ƒ†‹‹•–”ƒ–‹‘‘ˆƒƒƒ…–‹˜ƒ–‘”ȏ͹ȐǤ‘™‡˜‡”ǡƒ–’”‡•‡–ǡ‘‡˜‹†‡…‡Šƒ†•Š‘™–Šƒ–ƒ ™ƒ•‡š’”‡••‡†‹…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ –Š‹••–—†›ǡ‘—””‡•—Ž–•†‡‘•–”ƒ–‡–Šƒ–ƒ‹•’”‡•‡–‹”ƒ–…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ‡ ‹†‡–‹ϐ‹‡†„‘–Š–Š‡’”‘–‡‹Ž‡˜‡Ž•ƒ†–Š‡ˆ—…–‹‘ƒŽ”‡‰—Žƒ–‹‘‘ˆƒ‹…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ Materials and Methods Materials •‘’”‘–‡”‡‘ŽȋȌǡ…ƒŽ‹†‘Žǡ…ƒŽŠ‡šʹ͵ͳǡ•‘†‹—–ƒ—”‘—”•‘†‡‘š›…Š‘Žƒ–‡™‡”‡’—”…Šƒ•‡†ˆ”‘‹‰ƒǦ Ž†”‹…Šȋ–Ǥ‘—‹•ǡǡȌǤ–‹„‘†‹‡•ˆ‘”ƒǡǦʹǡǦ͵ǡǦͻǡǦ͵ǡǦͶǡȽǦƒ†  ™‡”‡ˆ”‘ƒ–ƒ”—œ‹‘–‡…Š‘Ž‘‰›Ǥ•‡…‘†ƒ”›ƒ–‹„‘†›™ƒ•Ǧ‰ ȋ”‘‡‰ƒȌǤ‘Ž›˜‹›Ž‹†‡‡†‹ϐŽ—‘”‹†‡ ȋ Ȍ‡„”ƒ‡•™‡”‡ˆ”‘Šƒ–ƒȋ‘™’ƒ”–‘ˆ ‡ƒŽ–Š…ƒ”‡‹ˆ‡…‹‡…‡•ǡ—…‹‰Šƒ•Š‹”‡ǡȌǤ ‡‘ƒ–ƒŽ”ƒ–…ƒ”†‹ƒ…ϔ‹„”‘„Žƒ•–•‹…—„ƒ–‹‘ ‡‘ƒ–ƒŽ ”ƒ– …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• ™‡”‡ ’”‡’ƒ”‡† ˆ”‘ ʹǦ –‘ ͵Ǧ†ƒ›Ǧ‘Ž† ‡‘ƒ–ƒŽ ‹•–ƒ” ”ƒ–• ȋ‹ƒŽ ‡•‡ƒ”…Š •–‹–—–‡ ‘ˆ ƒ”„‹ ‡†‹…ƒŽ ‹˜‡”•‹–›ǡ Š‹ƒȌǤ Š‡ ”ƒ–• ™‡”‡ ƒ‡•–Š‡–‹œ‡† ƒ† •ƒ…”‹ϐ‹…‡† „› ‹‡”•‹‘ ‹–‘ ͹ͷΨ ȋ˜Ȁ˜Ȍ ƒŽ…‘Š‘ŽǤ Š‡ ˜‡–”‹…Ž‡• ™‡”‡ ”‡‘˜‡† ƒ† ™ƒ•Š‡† –Š”‡‡ –‹‡• ‹ Ǧƒ• „ƒŽƒ…‡†•ƒŽ–•‘Ž—–‹‘ȋ‘ŽȀǣͲǤͶŽǡͲǤͲ͸ʹͶǡͺǤͲƒŽǡͲǤ͵ͷƒ͵ǡƒ†ͲǤͲ͸ƒʹͶ͹ʹǡ’ ͹ǤʹȌƒ–Ͷιǡ–Š‡‹…‡†ƒ†‹…—„ƒ–‡†™‹–ŠͲǤʹͷΨȋ™Ȁ˜Ȍ–”›’•‹ˆ‘”ͳͲ‹ƒ–͵͹ιǤ‘–‡”‹ƒ–‡–Š‡ †‹‰‡•–‹‘ǡ ƒ ‡“—ƒŽ ˜‘Ž—‡ ‘ˆ …‘Ž† —Ž„‡……‘ǯ• ‘†‹ϐ‹‡† ƒ‰Ž‡ǯ• ‡†‹— ȋȌ …‘–ƒ‹‹‰ ͳͲ Ψ ȋ˜Ȁ˜Ȍ ‡™„‘”…ƒŽˆ•‡”—™ƒ•ƒ††‡†ǤŠ‡•—’‡”ƒ–ƒ–™ƒ•†‹•…ƒ”†‡†Ǥ‡ŽŽ•™‡”‡‹…—„ƒ–‡†™‹–Šˆ”‡•ŠͲǤʹͷΨ –”›’•‹ˆ‘”ͳͷ‹ƒ–͵͹ιǡƒ†–Š‡•—’‡”ƒ–ƒ–™ƒ•…‘ŽŽ‡…–‡†ǤŠ‡Žƒ––‡”†‹‰‡•–‹‘•–‡’™ƒ•”‡’‡ƒ–‡†ˆ‘—” –‹‡•Ǥ ‡ŽŽ• ‹ –Š‡ •—’‡”ƒ–ƒ– ™‡”‡ –Š‡ ‹•‘Žƒ–‡† „› …‡–”‹ˆ—‰ƒ–‹‘ ˆ‘” ͳ͵ ‹ ƒ– ʹͶͲͲ ‰Ȁ‹ ƒ– ”‘‘ –‡’‡”ƒ–—”‡Ǥ ‡ŽŽ• ™‡”‡ ”‡Ǧ•—•’‡†‡† ‹  …‘–ƒ‹‹‰ ʹͲ Ψ ȋ˜Ȁ˜Ȍ ‡™„‘” …ƒŽˆ •‡”—ǡ ͳͲͲ ȀŽ ’‡‹…‹ŽŽ‹ƒ†ͳͲͲ‰ȀŽ•–”‡’–‘›…‹ǡƒ†–Š‡™‡”‡…—Ž–—”‡†ƒ•‘‘Žƒ›‡”•ƒ–ƒ†‡•‹–›‘ˆͷέͳͲͶ…‡ŽŽ•Ȁ …ʹƒ–͵͹鋐ƒŠ—‹†‹ϐ‹‡†ƒ–‘•’Š‡”‡…‘–ƒ‹‹‰ͷΨȋ˜Ȁ˜ȌʹǤˆ–‡”ͳŠ‹…—„ƒ–‹‘‹ϐŽƒ••ƒ–͵͹ιǡ ‹ƒŠ—‹†‹ϐ‹‡†ƒ–‘•’Š‡”‡™‹–ŠͷΨʹǡ–Š‡—ƒ––ƒ…Š‡†…‡ŽŽ•™‡”‡†‹•…ƒ”†‡†ƒ†–Š‡ƒ––ƒ…Š‡†…‡ŽŽ•™‡”‡ …‘–‹—‘—•Ž›…—Ž–—”‡†ǤŠ‡‡†‹—™ƒ•…Šƒ‰‡†–™‘–‹‡•’‡”™‡‡Ǥ Experimental protocol Š”‡‡†ƒ›•ƒˆ–‡”„‡‹‰•‡‡†‡†ǡ–Š‡…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•™‡”‡†‹˜‹†‡†”ƒ†‘Ž›‹–‘ˆ‘—”‰”‘—’•ǣȋͳȌ …‘–”‘Ž‰”‘—’ǣ‘–”‡ƒ–‡–ǢȋʹȌƒ‰‹‘–‡•‹ȋ‰Ȍ ‰”‘—’ǣ’”‡Ǧ–”‡ƒ–‡†™‹–ŠͳͲͲ‘ŽȀ‰ ˆ‘ŽŽ‘™‡† „›–”‡ƒ–‡–™‹–ŠʹǤͷƒŽʹˆ‘”ʹͶŠǢȋ͵ȌƒŽ‹†‘ŽΪƒ‰‹‘–‡•‹ȋ‰Ȍ ‰”‘—’ǣ’”‡Ǧ‹…—„ƒ–‡†™‹–ŠͳͲͲ 558 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion ‘ŽȀ‰ ˆ‘ŽŽ‘™‡†„›–”‡ƒ–‡–™‹–Š͵Ɋ…ƒŽ‹†‘Žˆ‘”ʹͶŠǢȋͶȌƒŽŠ‡šʹ͵ͳΪƒ‰‹‘–‡•‹ȋ‰Ȍ ‰”‘—’ǣ ’”‡Ǧ‹…—„ƒ–‡†™‹–ŠͳͲͲ‘ŽȀ‰ ˆ‘ŽŽ‘™‡†„›–”‡ƒ–‡–™‹–ŠʹɊ…ƒŽŠ‡šʹ͵ͳˆ‘”ʹͶŠǤ —‘ϔŽ—‘”‡•…‡–ƒ†‹—‘Š‹•–‘…Š‡‹…ƒŽ•–ƒ‹‹‰ ƒ”ƒˆϐ‹Ǧ‡„‡††‡† –‹••—‡ •‡…–‹‘• ȋͲǤʹ ɊȌ ™‡”‡ †‡’ƒ”ƒˆϐ‹‹œ‡† ™‹–Š š›Ž‡‡• ƒ† ”‡Š›†”ƒ–‡† ™‹–Š ‰”ƒ†‡† ™ƒ•Š‡• ™‹–Š ‡–Šƒ‘Ž ˆ‘ŽŽ‘™‡† „› ’Š‘•’Šƒ–‡Ǧ„—ˆˆ‡”‡† •ƒŽ‹‡ ȋȌǤ Š‡ǡ ‡ƒ…Š •Ž‹…‡ ™ƒ• –”‡ƒ–‡† ™‹–Š͵ͲɊŽ͵Ψʹʹȋ”‡ƒ‰‡–Ȍǡ‹…—„ƒ–‡†ƒ–”‘‘–‡’‡”ƒ–—”‡ˆ‘”ʹͲ‹ƒ†™ƒ•Š‡†–™‹…‡™‹–ŠǤ Š‡ǡ͵ͲɊŽ‰‘ƒ–•‡”—ȋ”‡ƒ‰‡–Ȍ™ƒ•ƒ††‡†ǡˆ‘ŽŽ‘™‡†„›‹…—„ƒ–‹‘ƒ–”‘‘–‡’‡”ƒ–—”‡ˆ‘”ʹͲ‹ƒ† –™‘™ƒ•Š‡•™‹–ŠǤƒ…Š•Ž‹…‡™ƒ•‹…—„ƒ–‡†‹͵ͲɊŽ’”‹ƒ”›ƒ–‹„‘†›ȋ‘—•‡ƒ–‹Ǧ”ƒ–ƒ‘‘…Ž‘ƒŽ ƒ–‹„‘†›ǡͳǣʹͲͲ†‹Ž—–‹‘Ȍƒ†’Žƒ…‡†‹ƒ™‡–„‘šƒ–Ͷι‘˜‡”‹‰Š–Ǥˆ–‡”™ƒ•Š‹‰™‹–Šǡ–Š‡•Ž‹…‡•™‡”‡ ‹…—„ƒ–‡†‹͵ͲɊŽ„‹‘–‹›Žƒ–‡†’‘Ž›…Ž‘ƒŽ•‡…‘†ƒ”›ƒ–‹„‘†›ȋ”‡ƒ‰‡–Ȍƒ–”‘‘–‡’‡”ƒ–—”‡ˆ‘”͵Ͳ‹ǡ ˆ‘ŽŽ‘™‡† „› ™ƒ•Š‹‰ ™‹–Š Ǥ Š‡ †‹ƒ‹‘„‡œ‹†‹‡ ȋȌ ‡–Š‘† ™ƒ• —•‡† ˆ‘” …‘Ž‘” †‡˜‡Ž‘’‡–ǡ ˆ‘ŽŽ‘™‡† „› ™ƒ•Š‹‰ ™‹–Š –ƒ’ ™ƒ–‡”Ǥ Ž‹…‡• ™‡”‡ –Š‡ …‘—–‡”•–ƒ‹‡† ™‹–Š Š‡ƒ–‘š›Ž‹ǡ ‹…—„ƒ–‡† ‹ ƒ‘‹ƒǡ†‡Š›†”ƒ–‡†™‹–Š‰”ƒ†‹‡–‡–Šƒ‘Žǡ–”ƒ•’ƒ”‡–‹œ‡†™‹–Šš›Ž‡‡•ƒ†ϐ‹ƒŽŽ›•‡ƒŽ‡†™‹–Š‡—–”ƒŽ ‰—Ǥ‡ŽŽ•™‹–Š„”‘™•–ƒ‹‹‰’ƒ”–‹…Ž‡•‹–Š‡‹”…›–‘’Žƒ•ƒ†—…Ž‡—•™‡”‡†‡‘–‡†’‘•‹–‹˜‡—†‡”ƒŽ‹‰Š– ‹…”‘•…‘’‡Ǥ ›–‘’Žƒ•‹…’”‘–‡‹‹•‘Žƒ–‹‘ˆ”‘…ƒ”†‹ƒ…ϔ‹„”‘„Žƒ•–• ”‘–‡‹‹•‘Žƒ–‹‘ƒ†ƒƒŽ›•‹•™‡”‡’‡”ˆ‘”‡†ƒ•†‡•…”‹„‡†’”‡˜‹‘—•Ž›ȏͺȐǤ”‹‡ϐŽ›ǡ™Š‘Ž‡Ǧ…‡ŽŽŽ›•ƒ–‡• ™‡”‡…‘ŽŽ‡…–‡†‹Ž›•‹•„—ˆˆ‡”ȋ͸ʹǤͷ‘ŽȀ‘ˆ”‹•ǦŽǡʹ‘ŽȀ‘ˆǡʹǤ͵Ψƒ†ͳͲΨ‰Ž›…‡”‘Žȏ’ ͸ǤͺȐȌǡ™‹–Š ƒ ’”‘–‡ƒ•‡‹Š‹„‹–‘”‹š–—”‡Ȍǡ ƒ† –‘–ƒŽ ’”‘–‡‹ …‘–‡– ™ƒ• †‡–‡”‹‡†„› –Š‡ „‹…‹…Š‘‹‹… ƒ…‹†‡–Š‘†ȋ‹‡”…‡ȌǤ“—ƒŽƒ‘—–•‘ˆ’”‘–‡‹•ƒ’Ž‡•™‡”‡•‡’ƒ”ƒ–‡†„›Ǧ ƒ†–”ƒ•ˆ‡””‡†–‘ ‹–”‘…‡ŽŽ—Ž‘•‡—•‹‰•–ƒ†ƒ”†–‡…Š‹“—‡•Ǥ ‡ƒ•—”‡‡–‘ˆ‹–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—‹…ƒ”†‹ƒ…ϔ‹„”‘„Žƒ•–• ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•™‡”‡Ž‘ƒ†‡†™‹–ŠͳɊ‘ŽȀ Ž‘—ǦͶȀƒ–͵͹ιˆ‘”͵Ͳ‹—–‡•‹„—ˆˆ‡”‡† •ƒŽ‹‡ȋ‹‘ŽȀǣͳ͵ͲƒŽǡͷŽǡͳͲ‰Ž—…‘•‡ǡͳ‰ŽʹǡͳǤͲƒŽʹǡƒ†ʹͷȏ’͹ǤͶȐȌǡƒ†‰”‘—’•‘ˆͷ –‘ͺ…‡ŽŽ•™‡”‡‘‹–‘”‡†—•‹‰ƒ‹˜‡”–‡†Ž›’—•Ǧ͹Ͳ‹…”‘•…‘’‡Ǥ’‡…–”‘ϐŽ—‘”‘‡–”‹…‡ƒ•—”‡‡–• ™‡”‡…‘ŽŽ‡…–‡†—•‹‰–Š‡‡Ž–ƒ…ƒ›•–‡•’‡…–”‘ϐŽ—‘”‘‡–‡”ȋŠ‘–‘‡…Š‘Ž‘‰›Ȍǡ™Š‡”‡–Š‡ϐ‹‡Ž†™ƒ• ‡š…‹–‡†ƒ–Ͷͺͺǡ™‹–Š‡‹••‹‘ƒ–ͷ͵ͲȏͻȐǤ MTT Proliferation Assay ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• ȋͳšͳͲͷ …‡ŽŽ• ’‡” ™‡ŽŽȌ ™‡”‡ •‡‡†‡†‹ ͸Ǧ™‡ŽŽ ’Žƒ–‡• ƒ† –”‡ƒ–‡† ™‹–Š ‘” ™‹–Š‘—– †‹ˆˆ‡”‡– ”‡ƒ‰‡–• ˆ‘” Ͷͺ Š‘—”•Ǥ Š‡”‡ƒˆ–‡”ǡ  ȋȋ͵ǦȋͶǡͷǦ‹‡–Š›Ž–Š‹ƒœ‘ŽǦʹǦ›ŽȌǦʹǡͷǦ†‹’Š‡›Ž–‡–”ƒœ‘Ž‹— „”‘‹†‡Ȍȋͷ‰ȀȌ™ƒ•ƒ††‡†–‘‡ƒ…Š™‡ŽŽǡƒ†’Žƒ–‡•™‡”‡‹…—„ƒ–‡†ƒ–͵͹ιˆ‘”ͶȂ͸Š‘—”•ǤŠ‡‡†‹— ™ƒ•–Š‡”‡’Žƒ…‡†™‹–ŠͳͷͲɊ†‹‡–Š›Ž•—Žˆ‘š‹†‡ȋȌƒ†ƒ‰‹–ƒ–‡†ˆ‘”ͳͲ‹—–‡•Ǥ„•‘”„ƒ…‡™ƒ• ‡ƒ•—”‡†ƒ–ͶͻͲ—•‹‰ƒ‹…”‘’Žƒ–‡”‡ƒ†‡”Ǥ ‡ŽŽ‘—–‹‰ ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•ȋͳšͳͲͷ’‡”™‡ŽŽȌ™‡”‡•‡‡†‡†‹͸Ǧ™‡ŽŽ’Žƒ–‡•ƒ†–”‡ƒ–‡†™‹–Š‘”™‹–Š‘—–†‹ˆˆ‡”‡– ”‡ƒ‰‡–•ˆ‘”ƒ†‡•‹‰ƒ–‡†’‡”‹‘†ǤŠ‡…‡ŽŽ•™‡”‡–Š‡†‹‰‡•–‡†„›ͲǤʹͷΨ–”›’•‹ˆ‘”͸Ȃͺ‹—–‡•ƒ–͵͹ιǤ Š‡…‡ŽŽ—„‡”™ƒ•…‘—–‡†—•‹‰ƒŠ‡‘…›–‘‡–‡”Ǥ …”ƒ–…Š™‘—†”‡’ƒ‹”ƒ••ƒ› ƒ–…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•™‡”‡–”‡ƒ–‡†™‹–Š…ƒŽ…‹—ǡ…ƒŽ‹†‘Žƒ†…ƒŽŠ‡šʹ͵ͳǡƒ†–Š‡•—„Œ‡…–‡†–‘–Š‡ in vitro •…”ƒ–…Šƒ••ƒ›ƒ•†‡•…”‹„‡†’”‡˜‹‘—•Ž›ȏͳͲǡͳͳȐǤƒ‰‡•™‡”‡…ƒ’–—”‡†ƒ–ͲǡʹͶƒ†ͶͺŠ‘—”•ƒˆ–‡” –”‡ƒ–‡–—•‹‰’Šƒ•‡Ǧ…‘–”ƒ•–‹…”‘•…‘’›Ǥ Animals ƒŽ‡‹•–ƒ””ƒ–•ȋʹͲͲȂʹͷͲ‰Ȍ™‡”‡‘„–ƒ‹‡†ˆ”‘–Š‡š’‡”‹‡–ƒŽ‹ƒŽ‡–‡”‘ˆƒ”„‹‡†‹…ƒŽ ‹˜‡”•‹–›ȋƒ”„‹ǡ‡‘’Ž‡ǯ•‡’—„Ž‹…‘ˆŠ‹ƒȌǤŽŽƒ‹ƒŽ‡š’‡”‹‡–ƒŽ’”‘–‘…‘Ž•…‘’Ž‹‡†™‹–Š–Š‡ǮǮ —‹†‡ ˆ‘”–Š‡ƒ”‡ƒ†•‡‘ˆƒ„‘”ƒ–‘”›‹ƒŽ•ǯǯ’—„Ž‹•Š‡†„›–Š‡Š‹‡•‡ƒ–‹‘ƒŽ•–‹–—–‡•‘ˆ‡ƒŽ–ŠǤŠ‡ •–—†› ™ƒ• ƒ’’”‘˜‡† „› –Š‡ •–‹–—–‹‘ƒŽ ‹ƒŽ ‡•‡ƒ”…Š ‘‹––‡‡ ‘ˆ ƒ”„‹ ‡†‹…ƒŽ ‹˜‡”•‹–›Ǥ ŽŽ ƒ‹ƒŽ•™‡”‡Š‘—•‡†ƒ––Š‡ƒ‹ƒŽ…ƒ”‡ˆƒ…‹Ž‹–›‘ˆƒ”„‹‡†‹…ƒŽ‹˜‡”•‹–›ƒ–ʹͷι™‹–ŠͳʹȀͳʹŠŽ‹‰Š–Ȁ 559 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion †ƒ”…›…Ž‡•ƒ†™‡”‡ƒŽŽ‘™‡†ˆ”‡‡ƒ……‡••–‘‘”ƒŽ”ƒ–…Š‘™ƒ†™ƒ–‡”–Š”‘—‰Š‘—––Š‡•–—†›’‡”‹‘†Ǥƒ–• ™‡”‡”ƒ†‘Ž›ƒ••‹‰‡†–‘†‹ˆˆ‡”‡––”‡ƒ–‡–‰”‘—’•Ǥ š’‡”‹‡–ƒŽ‰”‘—’• ȾǦ†”‡‡”‰‹…•–‹—Žƒ–‹‘…ƒ„‡‹†—…‡†„›ƒŠ‹‰Š†‘•‡‘ˆ‹•‘’”‘–‡”‡‘ŽȋȌǡ‹Œ‡…–‡†•—„…—–ƒ‡‘—•Ž› ƒ• ”‡’‘”–‡† „‡ˆ‘”‡ ȏͳʹȐǤ ‹š–› ”ƒ–• ™‡”‡ ”ƒ†‘Ž› †‹˜‹†‡† ‹–‘ ˆ‘—” ‰”‘—’•Ǥ  ‰”‘—’ ͳ ȋα ͳͲȌǡ –Š‡ ”ƒ–• ™‡”‡ •—„…—–ƒ‡‘—•Ž› ‹Œ‡…–‡† ™‹–Š •ƒŽ‹‡ ˆ‘” Ͷ †ƒ›• ƒ• ƒ …‘–”‘ŽǤ  ‰”‘—’ ʹ ȋǡ α ͳͷȌǡ –Š‡ ”ƒ–• ™‡”‡ •—„…—–ƒ‡‘—•Ž›‹Œ‡…–‡†™‹–Šȋͳ͹Ͳ‰Ȁ‰Ȁ†‹•ƒŽ‹‡Ȍˆ‘”Ͷ†ƒ›•–‘‹†—…‡…ƒ”†‹ƒ…Š›’‡”–”‘’Š›Ǥ‰”‘—’ ͵ȋǦǡαͳͺȌǡ–Š‡”ƒ–•™‡”‡•—„…—–ƒ‡‘—•Ž›‹Œ‡…–‡†™‹–Š…ƒŽ‹†‘ŽȋͳͲɊ‘ŽȀ‰Ȁ†‹•ƒŽ‹‡Ȍˆ‘”Ͷ™‡‡• ƒˆ–‡”ƒ†‹‹•–”ƒ–‹‘‘ˆˆ‘”Ͷ†ƒ›•ǤƒŽ‹†‘Ž‹•ƒ•’‡…‹ϐ‹…ƒ…–‹˜ƒ–‘”‘ˆƒǤ‰”‘—’ͶȋǦǡαͳ͹Ȍǡ–Š‡”ƒ–• ™‡”‡•—„…—–ƒ‡‘—•Ž›‹Œ‡…–‡†™‹–Š…ƒŽŠ‡šʹ͵ͳȋͳͲɊ‘ŽȀ‰Ȁ†‹•ƒŽ‹‡Ȍˆ‘”Ͷ™‡‡•ƒˆ–‡”ƒ†‹‹•–”ƒ–‹‘‘ˆ ˆ‘”Ͷ†ƒ›•ǤƒŽŠ‡šʹ͵ͳ‹•ƒ•’‡…‹ϐ‹…‹Š‹„‹–‘”‘ˆƒǤ ‡‘†›ƒ‹…‡ƒ•—”‡‡– ƒ”†‹ƒ… ˆ—…–‹‘• ™‡”‡ ƒ••‡••‡† —•‹‰ ƒ ‡…Š‘…ƒ”†‹‘‰”ƒ’Š› •›•–‡ ȋ ͹ ͳͲǡ ȌǤ …Š‘…ƒ”†‹‘‰”ƒ•™‡”‡’‡”ˆ‘”‡†‘•‡ŽˆǦ„”‡ƒ–Š‹‰”ƒ–•—†‡”ƒ‡•–Š‡•‹ƒȋ‹–”ƒ’‡”‹–‘‡ƒŽ‹Œ‡…–‹‘‘ˆͳͲ Ψ…ŠŽ‘”ƒŽŠ›†”ƒ–‡ƒ–ͲǤ͵ŽȀͳͲͲ‰„‘†›™‡‹‰Š–ȌǤ‡ˆ–˜‡–”‹…—Žƒ”ȋȌ’ƒ”ƒ‡–‡”•‡ƒ•—”‡†‹…Ž—†‡‹Ž‡– ˜‡–”‹…—Žƒ”•‡’–ƒŽ†‡ˆ‡…–•ȋ†Ȍƒ†Ž‡ˆ–˜‡–”‹…—Žƒ”‡Œ‡…–‹‘ˆ”ƒ…–‹‘ȋ ȌǤŽŽ’ƒ”ƒ‡–‡”•”‡’”‡•‡––Š‡ ‡ƒ•‘ˆϐ‹˜‡…‘•‡…—–‹˜‡…ƒ”†‹ƒ……›…Ž‡•ȏͳ͵ȐǤ ƒ••‘ǯ••–ƒ‹‹‰ ‡ƒ”–•ˆ”‘†‹ˆˆ‡”‡–‰”‘—’•ǡϐ‹š‡†‹ͳͲΨˆ‘”ƒŽ‹ǡ™‡”‡’”‘…‡••‡†ƒ†’ƒ”ƒˆϐ‹‡„‡††‡†Ǥ‡ƒ”– •‡…–‹‘• ȋͶ Ȍ ™‡”‡ –Š‡ •–ƒ‹‡† ™‹–Š ƒ••‘ǯ• –”‹…Š”‘‡ ”‡ƒ‰‡– ȏͳͶȐǤ Š‡ ’‡”…‡– ‘ˆ ϐ‹„”‘•‹• ȋ„Ž—‡ •–ƒ‹‹‰Ȍ–‘–‘–ƒŽ–‹••—‡™ƒ•ƒƒŽ›œ‡†ƒ†…ƒŽ…—Žƒ–‡†„›–Š‡Ǧ“—ƒ–‹–ƒ–‹˜‡ƒ—–‘ƒ–‹…’”‘‰”ƒ ȋ‹‘ǡ‘›‘ǡ ƒ’ƒȌǡ™‹–Š–Š‡ƒ˜‡”ƒ‰‡˜ƒŽ—‡”‡’”‡•‡–‹‰ƒ–Ž‡ƒ•–ͺ‹ƒ‰‡•’‡”Š‡ƒ”–ǡƒƒŽ›œ‡†‹†‘—„Ž‡Ǧ „Ž‹†ˆƒ•Š‹‘Ǥ‡”‹ƒŽǡ–”ƒ•˜‡”•‡…”›‘•‡…–‹‘•ȋ͹–Š‹…Ȍ‘ˆŠ‡ƒ”–•™‡”‡…—–™‹–Š–Š‡ͳͻͷͲ ”‹‰‘…—– ȋ‡‹…ƒǡ‡–œŽƒ”ǡ ‡”ƒ›Ȍƒ–ǦʹͲ郐†™‡”‡•–‘”‡†ƒ–ǦͺͲιǤ Statistical analysis ƒ–ƒƒ”‡‡š’”‡••‡†ƒ•–Š‡‡ƒάǤ‡Ǧ™ƒ›ƒƒŽ›•‹•‘ˆ˜ƒ”‹ƒ…‡ȋȌ™ƒ•—•‡†–‘†‡–‡”‹‡ •–ƒ–‹•–‹…ƒŽ•‹‰‹ϐ‹…ƒ…‡„‡–™‡‡‰”‘—’•ƒ†ƒŽ•‘–Š‡–™‘Ǧ–ƒ‹Ž‡†–—†‡–ǯ•–Ǧ–‡•–Ǥ’”‘„ƒ„‹Ž‹–›˜ƒŽ—‡‘ˆ’δͲǤͲͷ ™ƒ•–ƒ‡–‘‹†‹…ƒ–‡•–ƒ–‹•–‹…ƒŽ•‹‰‹ϐ‹…ƒ…‡Ǥ Results ”‘–‡‹‡š’”‡••‹‘‘ˆƒ‹”ƒ–‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϔ‹„”‘„Žƒ•–• ‘ †‡–‡”‹‡ ™Š‡–Š‡” ƒ ‹• ‡š’”‡••‡† ‹ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–•ǡ ƒ ‡š’”‡••‹‘ ™ƒ• †‡–‡…–‡† „› ‹—‘ϐŽ—‘”‡•…‡…‡ ƒ† ‹—‘Š‹•–‘…Š‡‹…ƒŽ •–ƒ‹‹‰Ǥ  ‡‘ƒ–ƒŽ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–•ǡ †‡‡’ ›‡ŽŽ‘™ ‹—‘•–ƒ‹‹‰ ‘” ‰”‡‡ ϐŽ—‘”‡•…‡…‡ ™ƒ• ’”‡•‡– –Š”‘—‰Š‘—– ƒŽŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•‹†‹…ƒ–‹‰–Š‡‡š’”‡••‹‘‘ˆƒƒ––Š‡’”‘–‡‹Ž‡˜‡Ž‹–Š‡•‡…‡ŽŽ•Ǥ Žƒ…‘ˆ•’‡…‹ϐ‹…•–ƒ‹‹‰™ƒ•†‡‘•–”ƒ–‡†‹–Š‡…‘–”‘Ž‰”‘—’ǡ‹–Š‡ƒ„•‡…‡‘ˆƒ–‹Ǧƒ ƒ–‹„‘†›ȋ ‹‰Ǥͳƒƒ†„ȌǤ Š‡ ‡š’”‡••‹‘ ‘ˆ ƒ ’”‘–‡‹ ™ƒ• ƒŽ•‘ ‡šƒ‹‡† ‹ ‡‘ƒ–ƒŽ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• „› ™‡•–‡”„Ž‘––‹‰Ǥƒ’”‘–‡‹•™‹–Šƒ”‡Žƒ–‹˜‡‘Ž‡…—Žƒ”ƒ••„‡–™‡‡ͳʹͲǦͳͶͲƒ™‡”‡ †‡–‡…–‡†‹‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•ȋ ‹‰Ǥͳ…ȌǤ —”–Š‡”‘”‡ǡ’”‡ƒ†•‘”’–‹‘‘ˆƒ–‹Ǧƒ ƒ–‹„‘†›™‹–Š•–ƒ†ƒ”†ƒƒ–‹‰‡‡Ž‹‹ƒ–‡†–Š‡ͳ͵Ͳƒ„ƒ†Ǥ‘‰‡–Š‡”ǡ–Š‡•‡”‡•—Ž–• ‹†‹…ƒ–‡–Šƒ–ƒ‹•‡š’”‡••‡†‹‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ …–‹˜ƒ–‹‘‘ˆƒ‡Ž‡˜ƒ–‡†–Š‡‹–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—…‘…‡–”ƒ–‹‘–Š”‘—‰Š–Š‡Ǧ͹ ’ƒ–Š™ƒ› ‘ …‘ϐ‹” –Šƒ– ƒ ˆ—…–‹‘ƒŽŽ› ”‡‰—Žƒ–‡• ‹–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹— …‘…‡–”ƒ–‹‘ ‹ ‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•ǡ…‡ŽŽ•™‡”‡–”‡ƒ–‡†™‹–ŠʹǤͷƒŽʹǡ‹ŽʹȋͳȌǡƒ‹Š‹„‹–‘”‘ˆ –Š‡ƒΪǦƒʹΪ‡š…Šƒ‰‡”ǡƒ††ŽʹȋʹͲͲɊȌǡƒ‹Š‹„‹–‘”‘ˆ–Š‡Ǧ–›’‡…ƒŽ…‹—…Šƒ‡Žǡ‹–Š‡ 560 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion Fig 1.ƒ‹•‡š’”‡••‡†‹‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•ǤȋƒȌ—‘ϐŽ—‘”‡•…‡…‡•–ƒ‹‹‰ˆ‘”ƒȋ‰”‡‡Ȍ ƒ†ȋ„Ž—‡ȌǤȋ„Ȍ—‘Š‹•–‘…Š‡‹•–”›ˆ‘”ƒȋ›‡ŽŽ‘™Ȍ’”‘–‡‹‡š’”‡••‹‘‹‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘Ǧ „Žƒ•–•ǤŠ‡„ƒ”‰”ƒ’Š•Š‘™•–Š‡“—ƒ–‹ϐ‹…ƒ–‹‘‘ˆ–Š‡ƒ”‡ƒ•‘ˆ’‘•‹–‹˜‡…‡ŽŽ•ȋ”‹‰Š–’ƒ‡ŽǢαͺȌǤƒ–ƒƒ”‡’”‡Ǧ •‡–‡†ƒ•–Š‡‡ƒάǤƒ‰‹ϐ‹…ƒ–‹‘ǣέͶͲͲǤȋ…Ȍ‡•–‡”„Ž‘––‹‰ƒƒŽ›•‹•‘ˆƒ‡š’”‡••‹‘‹‡‘ƒ–ƒŽ …ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ Fig 2. ƒ ƒ…–‹˜ƒ–‹‘ ‹†—…‡• ƒʹΪ ”‡Ž‡ƒ•‡ˆ”‘–Š‡‹‡‘ƒ–ƒŽ…ƒ”Ǧ †‹ƒ… ϐ‹„”‘„Žƒ•–•Ǥ ȋƒȌ ƒ ƒ…–‹˜ƒ–‹‘ ‹†—…‡† ƒʹΪ ”‡Ž‡ƒ•‡ ˆ”‘ –Š‡  ™‹–Š–Š‡–”‡ƒ–‡–‘ˆʹǤͷƒŽʹǤ ȋ„Ȍ ƒŽ‹†‘Ž ‹†—…‡† ƒʹΪ ”‡Ž‡ƒ•‡ ˆ”‘ –Š‡ Ǥ Š‡ …Šƒ‰‡ ‹ ϐŽ—‘”‡Ǧ •…‡…‡ ‹–‡•‹–›ǡ …‘””‡•’‘†‹‰ –‘ ‹–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹— …‘…‡–”ƒ–‹‘ǡ ™ƒ• ‡ƒ•—”‡† „› Ž—‘ǦͶ  •–ƒ‹Ǧ ‹‰ǡȋαͶǦͷȌǤȗ’δͲǤͲͷ˜•Ǥ…‘–”‘ŽǤ „ƒ–Š•‘Ž—–‹‘ǤŠ‡•‡–”‡ƒ–‡–•‹†—…‡†ƒ–‹‡Ǧ†‡’‡†‡–‹…”‡ƒ•‡‹‹–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—Ǥ ‹‹Žƒ”Ž›ǡ ƒ†‹‹•–”ƒ–‹‘ ‘ˆ ͳͲ Ɋ …ƒŽ‹†‘Ž ‡˜‘‡† ƒ ‹…”‡ƒ•‡ ‹ ‹–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹— …‘…‡–”ƒ–‹‘ ‹ ‘”‡ –Šƒ ͻͷ Ψ ‘ˆ ‡‘ƒ–ƒŽ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• ‹ ƒ ‰‹˜‡ ‘„•‡”˜ƒ–‹‘ ϐ‹‡Ž†Ǥ ˆ–‡” ’”‡–”‡ƒ–‡– ™‹–Š …ƒŽŠ‡šʹ͵ͳǡ ƒ •’‡…‹ϐ‹… ‹Š‹„‹–‘” ‘ˆ ƒǡ ‹–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹— …‘…‡–”ƒ–‹‘™ƒ•‘–•‹‰‹ϐ‹…ƒ–Ž›‹…”‡ƒ•‡†…‘’ƒ”‡†–‘ƒ†‹‹•–”ƒ–‹‘‘ˆ…ƒŽ‹†‘ŽƒŽ‘‡Ǥ ‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•™‡”‡’”‡–”‡ƒ–‡†ˆ‘”ͳͲ‹™‹–ŠͳͲɊ ȋ–Šƒ’•‹‰ƒ”‰‹Ȍǡ ™Š‹…Š ‹Š‹„‹–• ”‡ϐ‹ŽŽ‹‰ ‘ˆ –Š‡ ͵Ǧ•‡•‹–‹˜‡ …ƒŽ…‹— ”‡Ž‡ƒ•‡ ’‘‘Ž•Ǥ —„•‡“—‡–Ž›ǡ …‡ŽŽ• 561 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion Fig 3. ƒ ‹  † — … ‡ † …ƒ”†‹ƒ… ϐ‹Ǧ „ ” ‘ „ Ž ƒ • – ’ ”‘ Ž ‹ ˆ‡ ”ƒ Ǧ –‹‘ ƒ† ‹‰”ƒ–‹‘Ǥ ‡ŽŽ ’”‘Ǧ Ž ‹ ˆ ‡ ” ƒ – ‹ ‘  ‹†—…‡† „› ƒ…–‹˜ƒ–‹‘ ‘ˆ ƒǡ ƒ• ‡˜‹†‡…‡† „› ȋƒȌ …‡ŽŽ ˜ ‹ ƒ „ ‹ Ž ‹ – › ƒ† ȋ„Ȍ …‡ŽŽ  —  „ ‡ ” Ǥ ȋ…Ȍ …”ƒ–…Š ™‘—† ”‡Ǧ ’ƒ‹” ƒ••ƒ› ˆ‘” ‘„•‡”Ǧ ˜ƒ–‹‘ ‘ˆ  ‡ ‘  ƒ – ƒ Ž …ƒ”†‹ƒ… ϐ‹„Ǧ ”‘„Žƒ•– ‹Ǧ ‰”ƒ–‹‘Ǥ ™‡”‡ –”‡ƒ–‡† ™‹–Š ʹǤͷ  ƒŽʹ ƒ† ͳͲ Ɋ …ƒŽ‹†‘Žǡ ™Š‹…Š ˆƒ‹Ž‡† –‘ ‡Ž‹…‹– ƒ› ‹…”‡ƒ•‡ ‘ˆ ‹–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹— …‘…‡–”ƒ–‹‘Ǥ ”‡‹…—„ƒ–‹‘ ‘ˆ ‡‘ƒ–ƒŽ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• ˆ‘” ͳͲ ‹ ™‹–Š ͹͵ͳʹʹǡ ƒ ’Š‘•’Šƒ–‹†›Ž‹‘•‹–‘ŽǦ•’‡…‹ϐ‹…  „Ž‘…‡”ǡ ƒ„‘Ž‹•Š‡† –Š‡ ‹…”‡ƒ•‡ ‹ ‹–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—‹†—…‡†„›ͳͲɊ…ƒŽ‹†‘Ž‘”ʹǤͷƒŽʹǤ”‡–”‡ƒ–‡–™‹–ŠʹǦǡ ƒ͵”‡…‡’–‘”Ǧ•’‡…‹ϐ‹…‹Š‹„‹–‘”ǡˆ‘”ͳͲ‹‡Ž‹‹ƒ–‡†–Š‡‡ˆˆ‡…–‘ˆͳͲɊ…ƒŽ‹†‘Žƒ†ʹǤͷ ƒŽʹ‘–Š‡‹†—…–‹‘‘ˆ‹–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—”‡Ž‡ƒ•‡ȋ ‹‰ǤʹȌǤŠ‡•‡”‡•—Ž–•†‡‘•–”ƒ–‡ –Šƒ– –Š‡ ƒ…–‹˜ƒ–‹‘ ‘ˆ ƒ ”‡•—Ž–‡† ‹ ‹–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹— ‹…”‡ƒ•‡ ‹ ‡‘ƒ–ƒŽ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–•–Š”‘—‰Š•–‹—Žƒ–‹‘‘ˆ–Š‡Ǧ͵’ƒ–Š™ƒ›Ǥ ˆˆ‡…–•‘ˆƒ‘…‡ŽŽ’”‘Ž‹ˆ‡”ƒ–‹‘ƒ†‹‰”ƒ–‹‘‹‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϔ‹„”‘„Žƒ•–• ‘‹˜‡•–‹‰ƒ–‡™Š‡–Š‡”‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–’”‘Ž‹ˆ‡”ƒ–‹‘‹•”‡‰—Žƒ–‡†„›ƒǡ–Š‡ …‡ŽŽ•™‡”‡•–ƒ”˜‡†‘˜‡”‹‰Š–ƒ†’”‡–”‡ƒ–‡†™‹–Šƒƒ…–‹˜ƒ–‘”•ǡ‡‹–Š‡”ƒŽʹȋʹǤͷȌ‘” …ƒŽ‹†‘ŽȋͳͲɊȌǡˆ‘”ͳŠ‘—”ƒ†–Š‡‹…—„ƒ–‡†™‹–Š‰ ˆ‘”ͶͺŠ‘—”•ǤŠ‡”‡•—Ž–•‹†‹…ƒ–‡† –Šƒ– „‘–Š ƒŽʹ ƒ† …ƒŽ‹†‘Ž ‡Šƒ…‡† …‡ŽŽ †‡•‹–›ǡ ™Š‡”‡ƒ• –Š‡ ƒ „Ž‘…‡” …ƒŽŠ‡šʹ͵ͳ ƒ„‘Ž‹•Š‡†–Š‡•‡‡ˆˆ‡…–•Ǥ••Š‘™‹ ‹‰—”‡•͵ƒƒ†„ǡƒŽʹƒ†…ƒŽ‹†‘Ž‹†—…‡†…ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•– ’”‘Ž‹ˆ‡”ƒ–‹‘ǡ ƒ• ‡˜‹†‡…‡† „› …‡ŽŽ ˜‹ƒ„‹Ž‹–› ȋ ‹‰Ǥ ͵ƒȌ ƒ† ‹…”‡ƒ•‡† …‡ŽŽ —„‡” ȋ ‹‰Ǥ͵„ȌǤ ‘ƒ††”‡••™Š‡–Š‡”–Š‡‹‰”ƒ–‹‘‘ˆ‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–‹•‹†—…‡†„›ƒǡ…‡ŽŽ ‹‰”ƒ–‹‘ ™ƒ• •–—†‹‡† ‹ ƒ ™‘—†ǦŠ‡ƒŽ‹‰ ƒ••ƒ›Ǥ ‡ŽŽ• ‹ …—Ž–—”‡ ™‡”‡ •…”ƒ’‡† ‘ˆˆ ™‹–Š ƒ ’‹’‡––‡–‹’ǡ’”‘†—…‹‰ƒ™‹†‡ƒ…‡ŽŽ—Žƒ”ƒ”‡ƒǤƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•‹‰”ƒ–‹‰‹–‘–Š‹•ƒ…‡ŽŽ—Ǧ Žƒ”ƒ”‡ƒ™‡”‡…‘—–‡†ƒ†‡š’”‡••‡†ƒ•–Š‡—„‡”‘ˆ‹‰”ƒ–‡†…‡ŽŽ•Ǥ”‡ƒ–‡–™‹–ŠʹǤͷ ƒŽʹƒ†ͳͲɊ…ƒŽ‹†‘Ž•‹‰‹ϐ‹…ƒ–Ž›‹…”‡ƒ•‡†–Š‡‹‰”ƒ–‹‘‘ˆ‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘Ǧ „Žƒ•–•ƒˆ–‡”ʹͶŠ‹…—„ƒ–‹‘ǡ™Š‹Ž‡ƒ†‹‹•–”ƒ–‹‘‘ˆ…ƒŽŠ‡šʹ͵ͳ”‡†—…‡†–Š‹•‡ˆˆ‡…–ȋ ‹‰Ǥ͵…ȌǤ 562 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion Fig Š‡‡ˆˆ‡…–‘ˆƒ‘‡š–”ƒ…‡ŽŽ—Žƒ”ƒ–”‹š’”‘†—…–‹‘Ǥš’”‡••‹‘‘ˆȋƒȌƒǡȋ„ȌǦʹǡȋ…ȌǦͻǡ ȋ†ȌǦͶƒ†ȋ‡ȌƒǦ’”‘–‡‹•ǡƒ•‹†—…‡†„›–Š‡ƒ…–‹˜ƒ–‘”•…ƒŽ‹†‘Žƒ†ʹǤͷƒŽʹǤ”‘–‡‹Ž‡˜‡Ž• ™‡”‡‘”ƒŽ‹œ‡†–‘ ȋαͶǦͷȌǤȗ’δͲǤͲͷ˜•Ǥ…‘–”‘Ž‰”‘—’ǡ͓’δͲǤͲͷ˜•ǤʹǤͷƒŽʹ‰”‘—’Ǥ ˆˆ‡…–•‘ˆƒ‘‡š–”ƒ…‡ŽŽ—Žƒ”ƒ–”‹šȋȌ’”‘†—…–‹‘‹‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϔ‹„”‘„Žƒ•–• ‘ ‹˜‡•–‹‰ƒ–‡ ƒ ”‡‰—Žƒ–‹‰  ’”‘†—…–‹‘ǡ ƒ ‡š’”‡••‹‘ ™ƒ• †‡–‡…–‡†Ǥ  –Š‡ …ƒŽ‹†‘Ž ƒ† ƒŽʹ ‰”‘—’•ǡ ƒ ‡š’”‡••‹‘ ™ƒ• —’”‡‰—Žƒ–‡† …‘’ƒ”‡† –‘ …‘–”‘Ž ƒ† …ƒŽǦ Š‡šʹ͵ͳǦ–”‡ƒ–‡†‰”‘—’ȋ ‹‰ǤͶƒȌǤ 563 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion Fig 5.˜‘Ž˜‡‡–‘ˆƒ‹–Š‡ˆ—…–‹‘ƒŽ…Šƒ‰‡•‘ˆǦ‹†—…‡†…ƒ”†‹ƒ…Š›’‡”–”‘’Š›ǤȋƒȌƒ†ȋ„Ȍƒ ‹†—…‡†…Šƒ‰‡•‹›‘…ƒ”†‹ƒŽˆ—…–‹‘‹…ƒ”†‹ƒ…Š›’‡”–”‘’Š›ǡƒ••Š‘™„›ƒƒŽ›•‹•‘ˆ–Š‡Ž‡ˆ–˜‡–”‹…—Žƒ” ‡Œ‡…–‹‘ˆ”ƒ…–‹‘ȋ Ȍƒ†–Š‡‹–‡”˜‡–”‹…—Žƒ”•‡’–ƒŽ†‹ƒ•–‘Ž‹…–Š‹…‡••ȋ†ȌǤȋ…Ȍ‡’”‡•‡–ƒ–‹˜‡ƒ•Ǧ •‘–”‹…Š”‘‡•–ƒ‹‹‰‘ˆŠ‡ƒ”–•‡…–‹‘•ˆ”‘Ǧ‹†—…‡†…ƒ”†‹ƒ…Š›’‡”–”‘’Š›ǤŠ‡„ƒ”‰”ƒ’Š•Š‘™•–Š‡ “—ƒ–‹ϐ‹…ƒ–‹‘‘ˆ–Š‡ϐ‹„”‘–‹…ƒ”‡ƒ•‹Š‹•–‘Ž‘‰‹…ƒŽ•‡…–‹‘•ȋ”‹‰Š–’ƒ‡ŽǢαͺȌǤƒ‰‹ϐ‹…ƒ–‹‘ǣͶͲέʹͲͲǤ •‘–‘Ž›’Žƒ›ƒ”‘Ž‡‹ƒ–”‹š†‡‰”ƒ†ƒ–‹‘ǡ„—–ƒ”‡ƒŽ•‘‹˜‘Ž˜‡†‹…‘ŽŽƒ‰‡•›Ǧ –Š‡•‹• ”‡‰—Žƒ–‹‘Ǥ ‘ ”‡˜‡ƒŽ ™Š‡–Š‡” ƒ ‡ˆˆ‡…–‹‰ ‘  •‡…”‡–‹‘ǡ Ǧʹ ƒ† Ǧͻ ™‡”‡†‡–‡…–‡†„›‡•–‡”„Ž‘––‹‰Ǥ•‡š’‡…–‡†ǡͳͲɊ…ƒŽ‹†‘Žƒ†ʹǤͷƒŽʹ•‹‰‹ϐ‹Ǧ …ƒ–Ž›‡Šƒ…‡†–Š‡‡š’”‡••‹‘‘ˆǦʹƒ†Ǧͻ…‘’ƒ”‡†–‘…‘–”‘Žƒ†…ƒŽŠ‡šʹ͵ͳǦ –”‡ƒ–‡†‰”‘—’•ȋ ‹‰ǤͶ„ǡ…ȌǤǦͶ‡š’”‡••‹‘™ƒ••‹‰‹ϐ‹…ƒ–Ž›†‡…”‡ƒ•‡†‹…ƒŽ‹†‘Žƒ† ƒŽʹ‰”‘—’•ǡ„—–™ƒ•‹…”‡ƒ•‡†‹…ƒŽŠ‡šʹ͵ͳǦ–”‡ƒ–‡†‰”‘—’ȋ ‹‰ǤͶ†ȌǤ ȽǦȋƒŽ’ŠƒǦ•‘‘–Š—•…Ž‡ƒ…–‹Ȍ’‘•‹–‹˜‡ϐ‹„”‘„Žƒ•–•™‡”‡‹†‡–‹ϐ‹‡†ƒ•’”‹ƒ”›…‡ŽŽ –›’‡”‡•’‘•‹„Ž‡ˆ‘”‹–‡”•–‹–‹ƒŽƒ–”‹šƒ……——Žƒ–‹‘‹ϐ‹„”‘–‹…†‹•‡ƒ•‡•Ǥ—””‡•—Ž–••Š‘™‡† –Šƒ–ȽǦ‡š’”‡••‹‘™ƒ•‹…”‡ƒ•‡†‹–Š‡…ƒŽ‹†‘Žƒ†ƒŽʹ‰”‘—’•ȋ ‹‰ǤͶ‡ȌǤ ˆˆ‡…–• ‘ˆ ƒ ƒ…–‹˜ƒ–‹‘ ‘ …ƒ”†‹ƒ… ϔ‹„”‘•‹• ƒ† ›‘…ƒ”†‹ƒŽ ˆ—…–‹‘ ‹ Ǧ‹†—…‡† …ƒ”†‹ƒ…Š›’‡”–”‘’Š› ‘ˆ—”–Š‡”‹˜‡•–‹‰ƒ–‡–Š‡in vivo‡ˆˆ‡…–‘ˆƒ‘…ƒ”†‹ƒ…ϐ‹„”‘•‹•‹…ƒ”†‹ƒ…Š›’‡”–”‘’Š›ǡ ™‡ ‡•–ƒ„Ž‹•Š‡† ƒ ‘†‡Ž ‘ˆ …ƒ”†‹ƒ… Š›’‡”–”‘’Š› „› ‹Œ‡…–‹‰ ”ƒ–• ™‹–Š ƒ Š‹‰Š †‘•‡ ‘ˆ  ȋͳ͹Ͳ‰Ȁ‰Ȍˆ‘”Ͷ†ƒ›•Ǥƒ••‘ǯ••–ƒ‹‹‰ƒƒŽ›•‡••Š‘™‡†–Šƒ–…‘ŽŽƒ‰‡†‡’‘•‹–‹‘™ƒ• ‹…”‡ƒ•‡†‹–Š‡ǦͶ™ƒ†ǦͶ™Ǧ…ƒŽ‹†‘Ž‰”‘—’•ǡ…‘’ƒ”‡†™‹–Š–Š‡ǦͶ™Ǧ…ƒŽŠ‡šʹ͵ͳ ‰”‘—’•ȋ ‹‰Ǥͷ…ȌǤ 564 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion Fig 6.Š‡‡ˆˆ‡…–‘ˆƒ‘‡š–”ƒ…‡ŽŽ—Žƒ”ƒ–”‹š’”‘†—…–‹‘‹…ƒ”†‹ƒ…Š›’‡”–”‘’Š›ǤȋƒȌ‡•–‡”„Ž‘––‹‰ ƒƒŽ›•‹••Š‘™‹‰ƒ‡š’”‡••‹‘‹…ƒ”†‹ƒ…Š›’‡”–”‘’Š›‘†‡ŽǤš’”‡••‹‘‘ˆȋ„ȌǦʹǡȋ…ȌǦ͵ƒ† ȋ†ȌǦͻ’”‘–‡‹•‹†—…‡†„›–Š‡ƒ…–‹˜ƒ–‘”•…ƒŽ‹†‘ŽǤ”‘–‡‹Ž‡˜‡Ž•™‡”‡‘”ƒŽ‹œ‡†–‘ ȋαͶǦͷȌǤ ȗ’δͲǤͲͷ˜•Ǥ…‘–”‘Ž‰”‘—’ǡ͓’δͲǤͲͷ˜•ǤͶ™Ϊ…ƒŽ‹†‘Ž‰”‘—’Ǥ ‡‘†›ƒ‹… ’ƒ”ƒ‡–‡”•™‡”‡ ‘‹–‘”‡† „› ‡…Š‘…ƒ”†‹‘‰”ƒ’Š›Ǥ ‘’ƒ”‡† ™‹–Š –Š‡ …‘–”‘Ž‰”‘—’ǡ–Š‡‹–‡”˜‡–”‹…—Žƒ”•‡’–ƒŽ†‹ƒ•–‘Ž‹…–Š‹…‡••ȋ†Ȍ™ƒ•‹…”‡ƒ•‡†‹–Š‡Ǧ Ͷ™ǡǦͶ™Ǧ…ƒŽ‹†‘Žƒ†ǦͶ™Ǧ…ƒŽŠ‡šʹ͵ͳ‰”‘—’•ǤŠ‡Ž‡ˆ–˜‡–”‹…—Žƒ”‡Œ‡…–‹‘ˆ”ƒ…–‹‘™ƒ• †‡…”‡ƒ•‡†‹–Š‡ǦͶ™ƒ†ǦͶ™Ǧ…ƒŽ‹†‘Ž‰”‘—’•ƒˆ–‡”ƒ†‹‹•–”ƒ–‹‘ˆ‘”Ͷ™‡‡•ǡ …‘’ƒ”‡†™‹–Š–Š‡…‘–”‘Ž‰”‘—’ȋ ‹‰Ǥͷƒǡ„ȌǤ ‡ƒŽ•‘ˆ‘—†–Šƒ––Š‡‡š’”‡••‹‘‘ˆƒ‹Š‡ƒ”––‹••—‡•™ƒ••‹‰‹ϐ‹…ƒ–Ž›‹…”‡ƒ•‡† ‹–Š‡ǦͶ™ƒ†ǦͶ™Ǧ…ƒŽ‹†‘Ž‰”‘—’•ȋ ‹‰Ǥ͸ƒȌǤ‘‡š’Ž‘”‡–Š‡in vivo‡ˆˆ‡…–‘ˆƒ‘ 565 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion …ƒ”†‹ƒ… ϐ‹„”‘•‹• ‹ …ƒ”†‹ƒ… Š›’‡”–”‘’Š›ǡ ™‡ †‡–‡…–‡† –Š‡ ‡š’”‡••‹‘ ‘ˆ •‡˜‡”ƒŽ ‘Ž‡…—Žƒ” ‹†‹…ƒ–‘”•‘ˆ…ƒ”†‹ƒ…ϐ‹„”‘•‹•ǤŠ‡‡š’”‡••‹‘‘ˆ„‘–ŠǦʹǡ͵ƒ†Ǧͻ‹–Š‡ǦͶ™ƒ†Ǧ Ͷ™Ǧ…ƒŽ‹†‘Ž‰”‘—’•™ƒ•‹…”‡ƒ•‡†…‘’ƒ”‡†–‘–Š‡ǦͶ™Ǧ…ƒŽŠ‡šʹ͵ͳƒ†…‘–”‘Ž‰”‘—’•Ǥ Š‡•‡ ”‡•—Ž–• ˆ—”–Š‡” †‡‘•–”ƒ–‡ –Šƒ– ƒ…–‹˜ƒ–‹‘ ‘ˆ ƒ ‹• ‹˜‘Ž˜‡† ‹ –Š‡ ˆ‘”ƒ–‹‘ ‘ˆ …ƒ”†‹ƒ…ϐ‹„”‘•‹•‹Ǧ‹†—…‡†…ƒ”†‹ƒ…Š›’‡”–”‘’Š›ȋ ‹‰Ǥ͸„ǡ…ǡ†ȌǤ Discussion Š‡ ‡š’”‡••‹‘ ‘ˆ ƒ ‹ ˜ƒ”‹‘—• …‡ŽŽ• ‹• ”‡Žƒ–‡† –‘ —Ž–‹’Ž‡ …‡ŽŽ—Žƒ” ˆ—…–‹‘•ǡ •—…Š ƒ• …‡ŽŽ ’”‘Ž‹ˆ‡”ƒ–‹‘ǡ ƒ’‘’–‘•‹• ƒ† –Š‡ ”‡‰—Žƒ–‹‘ ‘ˆ •›•–‡‹… …ƒŽ…‹— Š‘‡‘•–ƒ•‹• ȏͳͷȐǤ ‘”‹•–ƒ…‡ǡ”‘™‡–ƒŽǤŠƒ˜‡…‘ϐ‹”‡†–Šƒ–ƒ‹•ƒ…–‹˜ƒ–‡†„›‡š–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—‹ ’ƒ”ƒ–Š›”‘‹† …‡ŽŽ• ƒ† ‹• ”‡•’‘•‹„Ž‡ ˆ‘” –Š‡ ”‡Ž‡ƒ•‡ ‘ˆ  ȏͳ͸ǦͳͺȐǤ —” ’”‡˜‹‘—• •–—†‹‡• †‡‘•–”ƒ–‡†–Šƒ––Š‡ƒ…–‹˜ƒ–‹‘‘ˆƒ‹†—…‡†…ƒ”†‹‘›‘…›–‡ƒ’‘’–‘•‹•†—”‹‰‹•…Š‡‹ƒȀ ”‡’‡”ˆ—•‹‘‹Œ—”›ƒ†‹ˆƒ‹Ž‹‰Š‡ƒ”–•ȏ͹ǡͺȐǤ—”’”‡•‡–•–—†›†‡‘•–”ƒ–‡•ǡˆ‘”–Š‡ϐ‹”•– –‹‡ǡ–Š‡‡š‹•–‡…‡‘ˆƒ‹”ƒ–…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•ǤŠ‹•…‘…Ž—•‹‘‹•„ƒ•‡†‘•‡˜‡”ƒŽŽ‹‡• ‘ˆ ‡˜‹†‡…‡ǣ ȋƒȌ ƒ ’”‘–‡‹• ™‡”‡ ‹†‡–‹ϐ‹‡† ‹ ‡‘ƒ–ƒŽ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• „› ‡•–‡” „Ž‘– ƒƒŽ›•‹•Ǣ ȋ„Ȍ ƒ ’”‘–‡‹• ™‡”‡ •Š‘™ –‘ „‡ Ž‘…ƒ–‡† ‹ ‡‘ƒ–ƒŽ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• „› ‹—‘Š‹•–‘Ž‘‰‹…ƒŽ ƒ† ‹—‘ϐŽ—‘”‡•…‡…‡ •–ƒ‹‹‰Ǣ ȋ…Ȍ ‡š–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹— ƒ† …ƒŽ‹†‘Žǡƒ•’‡…‹ϐ‹…ƒ…–‹˜ƒ–‘”‘ˆƒǡ‹…”‡ƒ•‡†‹–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—Ž‡˜‡Ž•ǡ–Š‡”‡„›ƒ…–‹˜ƒ–‹‰ –Š‡Ǧ͵’ƒ–Š™ƒ›ƒ†‹†—…‹‰–Š‡”‡Ž‡ƒ•‡‘ˆ…ƒŽ…‹—ˆ”‘Ǣȋ†Ȍ‡š–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹— ƒ† …ƒŽ‹†‘Ž ’”‘‘–‡† ‡‘ƒ–ƒŽ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•– ’”‘Ž‹ˆ‡”ƒ–‹‘ǡ  ‹‰”ƒ–‹‘ ƒ†  •‡…”‡–‹‘–Š”‘—‰Š–Š‡ƒ…–‹˜ƒ–‹‘ ‘ˆƒǢƒ† ȋ‡Ȍƒ’Žƒ›‡†ƒ ”‘Ž‡ ‹Ǧ‹†—…‡† …ƒ”†‹ƒ… ϐ‹„”‘•‹•‹”ƒ–•Ǥ CaʹΪ •‹‰ƒŽ• ƒ”‡ ™‡ŽŽ •–—†‹‡† ‹ …ƒ”†‹ƒ… ›‘…›–‡• ȏͳͻǡ ʹͲȐǤ ‘™‡˜‡”ǡ ƒʹΪ •‹‰ƒŽ‹‰ ’ƒ–Š™ƒ›•ƒ”‡‘–™‡ŽŽ—†‡”•–‘‘†‹‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ‹‡–ƒŽǤŠƒ˜‡”‡˜‡ƒŽ‡†–Šƒ– CaʹΪƒ…–‹˜‹–›‹Š—ƒ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•‹•‡†‹ƒ–‡†„›—Ž–‹’Ž‡ƒʹΪ•‹‰ƒŽ‹‰’ƒ–Š™ƒ›•ǡ ‹…Ž—†‹‰͵•ǡ•ǡǡ•ƒ†•ȏʹͳȐǤƒǡƒ•ƒ ’”‘–‡‹…‘—’Ž‡†”‡…‡’–‘”ǡ •‡•‡•…Šƒ‰‡•‹‡š–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—…‘…‡–”ƒ–‹‘ǡ–”‹‰‰‡”‹‰ƒ‹…”‡ƒ•‡‹‹–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹—ȏͳͺȐǤƒǡ‹„‘–Š”ƒ–…ƒ”†‹ƒ…›‘…›–‡•ƒ†Š—ƒƒ–”ƒŽ‰ƒ•–”‹…‡ŽŽ•ǡ‹••‡•‹–‹˜‡ –‘…Šƒ‰‡•‹‡š–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—…‘…‡–”ƒ–‹‘ȏͳͷǡʹʹȐǤ—””‡•—Ž–•‹‡‘ƒ–ƒŽ…ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–•ƒ”‡•‹‹Žƒ”–‘–Š‡”‡•—Ž–•†‡•…”‹„‡†ƒ„‘˜‡Ǥ Š‡ •ƒ”…‘’Žƒ•‹…Ȁ‡†‘’Žƒ•‹… ”‡–‹…—Ž—ǡ ƒ •’‡…‹ƒŽ‹œ‡† …ƒŽ…‹—Ǧ•–‘”‹‰ ‘”‰ƒ‡ŽŽ‡ǡ ‹• ™‡ŽŽ ‘™ –‘ „‡ ‹–‹ƒ–‡Ž› ‹˜‘Ž˜‡† ‹ ”‡‰—Žƒ–‹‰ ƒʹΪ ‘˜‡‡– ™‹–Š‹ …‡ŽŽ•Ǥ ͵• ƒ† ›• ’ƒ”–‹…‹’ƒ–‡ ‹ –Š‡ ”‡Ž‡ƒ•‡‘ˆƒʹΪ ˆ”‘ –Š‡ •ƒ”…‘’Žƒ•‹…Ȁ‡†‘’Žƒ•‹… ”‡–‹…—Ž— ȏʹ͵ǡ ʹͶȐǤ –‡”‡•–‹‰Ž›ǡ ‹ ‡– ƒŽǤ ˆ‘—† –Šƒ– ›• ƒ› ‘– „‡ ‹˜‘Ž˜‡† ‹ ƒʹΪ ƒ…–‹˜‹–› ‹ Š—ƒ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–•Ǥ Š‡•‡ ”‡•—Ž–• †‡‘•–”ƒ–‡† –Šƒ– ‹–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹— ‹ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• ƒ”‡ ‡†‹ƒ–‡† „› ͵• ȏʹͳȐǤ —” ”‡•—Ž–• ”‡˜‡ƒŽ –Šƒ– ‹…”‡ƒ•‡† ‹–”ƒ…‡ŽŽ—Žƒ” …ƒŽ…‹—…‘…‡–”ƒ–‹‘•™‡”‡‹Š‹„‹–‡†„›͹͵ͳʹʹƒ†ʹǦǤƒ††‹–‹‘ǡ–Š‡ƒƒ…–‹˜ƒ–‘” …ƒŽ‹†‘Ž‹†—…‡†•‹‰‹ϐ‹…ƒ–Ž›‹…”‡ƒ•‡†‹–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—…‘…‡–”ƒ–‹‘•ǤŠ‡•‡”‡•—Ž–• †‡‘•–”ƒ–‡–Šƒ–ƒ‹•‹˜‘Ž˜‡†‹‹…”‡ƒ•‹‰–Š‡‹–”ƒ…‡ŽŽ—Žƒ”ƒʹΪ…‘…‡–”ƒ–‹‘–Š”‘—‰Š –Š‡Ǧ͵’ƒ–Š™ƒ›Ǥ ƒ”†‹ƒ… ϐ‹„”‘•‹• ‹• ˜‹–ƒŽ ˆ‘” ˜‡–”‹…—Žƒ” ”‡‘†‡Ž‹‰ ȏʹͷȐǤ ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–•ǡ Š‹‰ŠŽ› ƒ…–‹˜‡…‡ŽŽ•–Šƒ–‡šŠ‹„‹–‹…”‡ƒ•‡†’”‘Ž‹ˆ‡”ƒ–‹˜‡ǡ‹‰”ƒ–‘”›ƒ†•‡…”‡–‘”›’”‘’‡”–‹‡•ȏʹ͸ǡʹ͹Ȑǡ ”‡’”‡•‡––Š‡Žƒ”‰‡•–…Žƒ••‘ˆ…‡ŽŽ•”‡•‹†‹‰‹–Š‡Š‡ƒ”–Ǥ”‘Ž‹ˆ‡”ƒ–‹‘‘ˆ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•‹• –Š‡ƒ‹…Šƒ”ƒ…–‡”‹•–‹…‘ˆ›‘…ƒ”†‹ƒŽϐ‹„”‘•‹•ǤŽ–Š‘—‰Š–Š‡‹’‘”–ƒ…‡‘ˆϐ‹„”‘•‹•‹˜ƒ”‹‘—• …ƒ”†‹ƒ…†‹•‡ƒ•‡•Šƒ•„‡‡™‡ŽŽ‡•–ƒ„Ž‹•Š‡†ǡ–Š‡ƒʹΪ•‹‰ƒŽ‹‰‡…Šƒ‹••‹˜‘Ž˜‡†‹…ƒ”†‹ƒ… ϐ‹„”‘•‹•ƒ”‡‘–‘™Ǥ‘‡•–—†‹‡•”‡˜‡ƒŽ‡†–Šƒ–͹‹•Ž‘…ƒ–‡†‹–Š‡‡„”ƒ‡‘ˆ …ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•ƒ†‡†‹ƒ–‡•Š‹‰Š‡”ȏƒʹΪȐi‘•…‹ŽŽƒ–‹‘ˆ”‡“—‡…‹‡•‹†‹…ƒ–‹˜‡‘ˆŠ‹‰Š‡” •–‹ˆˆ‡••ǡ™Š‹…Š™‘—Ž†”‡•—Ž–‹‘”‡‡ˆϐ‹…‹‡–”‡‘†‡Ž‹‰ȏʹͺǦ͵ͲȐǤƒʹڏƒ›’Žƒ› ƒ ‡••‡–‹ƒŽ ”‘Ž‡ ‹ …ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–• ’”‘Ž‹ˆ‡”ƒ–‹‘ ƒ† †‹ˆˆ‡”‡–‹ƒ–‹‘ ȏͳͻȐǤ —” ”‡•—Ž–• …‘ϐ‹”–Šƒ–ƒ…–‹˜ƒ–‹‘‘ˆƒ…‘—Ž†’”‘‘–‡‹‰”ƒ–‹‘ƒ†‡Ž‡˜ƒ–‡–Š‡‡š’”‡••‹‘Ž‡˜‡Ž• ‘ˆǦʹƒ†ͻ‹‡‘ƒ–ƒŽ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ‘ˆ—”–Š‡”†‡‘•–”ƒ–‡–Š‡‹˜‘Ž˜‡‡–‘ˆ 566 Cellular Physiology and Biochemistry Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion ƒ ‹ …ƒ”†‹ƒ… ϐ‹„”‘•‹•ǡ ƒ ”ƒ– ‘†‡Ž ‘ˆ Ǧ‹†—…‡† …ƒ”†‹ƒ… Š›’‡”–”‘’Š› ƒ† ϐ‹„”‘•‹• ™ƒ• ‡•–ƒ„Ž‹•Š‡†ǡˆ‘ŽŽ‘™‡†„›ƒ†‹‹•–”ƒ–‹‘‘ˆ…ƒŽ‹†‘Žƒ†…ƒŽŠ‡šʹ͵ͳǤƒ••‘ǯ••–ƒ‹‹‰•Š‘™‡† –Šƒ–…ƒŽ‹†‘Ž‡Šƒ…‡†…ƒ”†‹ƒ…ϐ‹„”‘•‹•ǡ™Š‡”‡ƒ•…ƒŽŠ‡šʹ͵ͳ†‹‹‹•Š‡†…ƒ”†‹ƒ…ϐ‹„”‘•‹•ǤŠ‡ ‡š’”‡••‹‘‘ˆ„‘–ŠǦʹƒ†Ǧͻ™ƒ•‡Ž‡˜ƒ–‡†„›–”‡ƒ–‡–™‹–Š…ƒŽ‹†‘ŽǤ Š‡ ƒǦ‡†‹ƒ–‡† –”‹‰‰‡”‹‰ ‘ˆ …ƒ”†‹ƒ… ϐ‹„”‘•‹• ˜‹ƒ ƒʹΪ •‹‰ƒŽ‹‰ ƒ› •‡”˜‡ ƒ• ƒ …‘‘’ƒ–Š™ƒ›‹–Š‡ϐ‹„”‘•‹•…ƒ•…ƒ†‡Ǥˆ•‘ǡ‹Š‹„‹–‹‘‘ˆƒʹΪ”‡Ž‡ƒ•‡ˆ”‘–Š‡–Š”‘—‰Š ƒƒ›ƒ––‡—ƒ–‡…ƒ”†‹ƒ…ϐ‹„”‘•‹•ǤŠ‡…‘–”‹„—–‹‘‘ˆƒǦ‡†‹ƒ–‡†ƒʹΪ•‹‰ƒŽ‹…ƒ”†‹ƒ… ϐ‹„”‘„Žƒ•–••Š‘™‹–Š‹••–—†›‹•–Š‡ϐ‹”•–†‡‘•–”ƒ–‹‘‘ˆƒ‹’‘”–ƒ–”‘Ž‡‘ˆ–Š‹•”‡…‡’–‘” ‹Š‡ƒ”–†‹•‡ƒ•‡Ǥ …‘…Ž—•‹‘ǡ‘—””‡•—Ž–•ƒ”‡–Š‡ϐ‹”•––‘”‡’‘”–ƒ‹’‘”–ƒ–”‘Ž‡ˆ‘”ƒ‹ƒʹΪ•‹‰ƒŽ‹‰ ‹˜‘Ž˜‡†‹…ƒ”†‹ƒ…ϐ‹„”‘•‹•ƒ†–Šƒ––Š‹•”‘Ž‡‹•‡†‹ƒ–‡†„›–Š‡Ǧ͵’ƒ–Š™ƒ›Ǥ—”•–—†› ‘’‡•—’ƒ‡™ƒ˜‡—‡–‘‡š’Ž‘”‡–Š‡’‘–‡–‹ƒŽ”‘Ž‡•‘ˆƒʹΪ•‹‰ƒŽ‹‰‹…ƒ”†‹ƒ…ϐ‹„”‘•‹•ƒ† …‘—Ž†Ž‡ƒ†–‘–Š‡†‡˜‡Ž‘’‡–‘ˆ‘”‡‡ˆˆ‡…–‹˜‡ƒ’’”‘ƒ…Š‡•ˆ‘”–”‡ƒ–‹‰…ƒ”†‹ƒ…ϐ‹„”‘•‹•Ǥ &RQÁLFWRI,QWHUHVW Š‡ƒ—–Š‘”•†‡…Žƒ”‡–Šƒ––Š‡›Šƒ˜‡‘…‘ϐŽ‹…–‘ˆ‹–‡”‡•–Ǥ Acknowledgments Š‹• •–—†› ™ƒ• •—’’‘”–‡† „› –Š‡ ƒ–‹‘ƒŽ ƒ–—”ƒŽ …‹‡…‡ ‘—†ƒ–‹‘ ‘ˆ Š‹ƒ ȋͺͳͳ͹Ͳʹͺͻǡ ͺͳͳ͹Ͳʹͳͺǡ ͺͳ͵͹ͲͶʹͳǡ ͺͳ͵͹Ͳ͵͵Ͳǡ ͺͳ͵ͲͲͳ͸Ͷǡ ͺͳͳ͹Ͳͳ͹ͺȌǡ –Š‡ ƒ”„‹ ‡†‹…ƒŽ ‹˜‡”•‹–›‰”ƒ–ˆ‘”‡š…‡ŽŽ‡–›‘—‰‡”•…‹‡–‹•–•ȋ–‘‡‹Š—ƒŠƒ‰Ȍƒ†Š‹ƒ‘•–†‘…–‘”ƒŽ …‹‡…‡ ‘—†ƒ–‹‘ ˆ—†‡† ’”‘Œ‡…– ȋʹͲͳʹͷʹͲ͹͸͸Ȍ ƒ† ‡‹Ž‘‰Œ‹ƒ‰ ‘•–†‘…–‘”ƒŽ —† ȋ‘ǤǦͳͳͲͺͶȌ ˆ‘” ƒ‰Šƒ‘ — ƒ† ƒ”„‹ …‹‡…‡ ƒ† –‡…Š‘Ž‘‰› „—”‡ƒ— —† ȋʹͲͲͻ ͲͳͶȌˆ‘”‹ƒƒ‰Ǥ Reference ƒ—†‹‘ǡƒ”˜‡”ǡ ‹Ž‡•ǡ‘”‰ǣƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•ǣ ”‹‡†‘” ‘‡ǫ Š›•‹‘Ž‡ƒ”–‹”…Š›•‹‘Ž ʹͲͲ͸ǢʹͻͳǣͳͲͳͷǦͳͲʹ͸Ǥ Š‡ ǡƒ‘ǡ—ǡ•‡ ǡƒ—ǡ‹ ǣ—Ž–‹’Ž‡ƒʹΪ•‹‰ƒŽ‹‰’ƒ–Š™ƒ›•”‡‰—Žƒ–‡‹–”ƒ…‡ŽŽ—Žƒ”ƒʹΪ ƒ…–‹˜‹–›‹Š—ƒ…ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ ‡ŽŽŠ›•‹‘ŽʹͲͳͲǢʹʹ͵ǣ͸ͺǦ͹ͷǤ ‘‘–ƒǡ‘ŽŽ‹• ǡƒ…‡œ‹‡ǡ‘†‡”‹…ǡ‡””‹†‰‡ ǡ‡’’‹ƒ––ǣʹǦƒ‹‘‡–Š‘š›†‹’Š‡›Ž „‘”ƒ–‡ȋʹǦȌ‹•ƒ”‡Ž‹ƒ„Ž‡„Ž‘…‡”‘ˆ•–‘”‡Ǧ‘’‡”ƒ–‡†ƒʹΪ‡–”›„—–ƒ‹…‘•‹•–‡–‹Š‹„‹–‘”‘ˆ•͵Ǧ ‹†—…‡†ƒʹΪ”‡Ž‡ƒ•‡Ǥ  ʹͲͲʹǢͳ͸ǣͳͳͶͷǦͳͳͷͲǤ ƒ‰ ǡ‘Šƒ”ǡ…—ŽŽ‘…Šǣ—Ž–‹’Ž‡”‘Ž‡•‘ˆƒŽ’ŠƒǦ•‘‘–Š—•…Ž‡ƒ…–‹‹‡…Šƒ‘–”ƒ•†—…–‹‘Ǥš’ ‡ŽŽ‡•ʹͲͲ͸Ǣ͵ͳʹǣʹͲͷǦʹͳͶǤ ‹ƒ‰ǡ…‘ƒŽ†ǡ…ƒ—•ǡ˜ƒ”‡‡‡ǡƒ‰ǣʹʹ”‡…‡’–‘”Ǧ‡†‹ƒ–‡†ƒʹΪ•‹‰ƒŽ‹‰ƒ† •’‘–ƒ‡‘—•ƒʹΪ”‡Ž‡ƒ•‡•‹Š—ƒ˜ƒŽ˜—Žƒ”›‘ϐ‹„”‘„Žƒ•–•ǡ–‡ƒ”– ʹͲͲͺǢͶͻǣʹʹͳȂʹ͵͸Ǥ ”‘™ǡƒ…‡‘† ǣš–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—•‡•‹‰ƒ†‡š–”ƒ…‡ŽŽ—Žƒ”…ƒŽ…‹—•‹‰ƒŽ‹‰ǤŠ›•‹‘Ž‡˜ ʹͲͲͳǢͺͳǣʹ͵ͻȂͻ͹Ǥ — ǡ —ǡ‡‰ǡŠƒ‰ǡ‘‰ǡŠƒ‘ ǡ‡ǡ‹ǡŠ‘‰ǡ—ǡŠƒ‰ǣ‘Ž‡‘ˆ–Š‡ ƒŽ…‹—Ǧ‡•‹‰‡…‡’–‘”‹ƒ”†‹‘›‘…›–‡’‘’–‘•‹•˜‹ƒ–Š‡ƒ”…‘’Žƒ•‹…‡–‹…—Ž—ƒ†‹–‘…Š‘†”‹ƒŽ ‡ƒ–Šƒ–Š™ƒ›‹ƒ”†‹ƒ…›’‡”–”‘’Š›ƒ†‡ƒ”– ƒ‹Ž—”‡Ǥ‡ŽŽŠ›•‹‘Ž‹‘…Š‡ʹͲͳ͵Ǣ͵ͳǣ͹ʹͺǦ͹Ͷ͵Ǥ —ǡƒ‰ǡƒŽ‘•ǡ‡”‡†‹ ǡ‹ǡƒ‰ǡ —›–‘ǡŠƒ‘ǣ˜‘Ž˜‡‡–‘ˆƒΪȀΪ ‡š…Šƒ‰‡”‹Š›’‘š‹ƒȀ”‡Ǧ‘š›‰‡ƒ–‹‘‹†—…‡†‡‘ƒ–ƒŽ”ƒ–…ƒ”†‹‘›‘…›–‡ƒ’‘’–‘•‹•Ǥ—” Šƒ”ƒ…‘Ž ʹͲͲͶǢͶͺ͸ǣͳʹͳǦͳ͵ͳǤ ”‹ǤŽ•‘ǡƒ–”‹…‹ƒǤŠƒŠƒ”–ǡƒ—‰Ž‡ ‡‹ˆ‡”Ǥƒ† ǣ ƒ”›‡•œƒ”‘•Ǥ‰‹‘–‡•‹ Ǧ†—…‡† š–”ƒ…‡ŽŽ—Žƒ”‹‰ƒŽǦ‡‰—Žƒ–‡†‹ƒ•‡ͳȀʹ…–‹˜ƒ–‹‘•‡†‹ƒ–‡†„›”‘–‡‹‹ƒ•‡Gƒ†–”ƒ…‡ŽŽ—Žƒ” ƒŽ…‹—‹†—Ž–ƒ–ƒ”†‹ƒ… ‹„”‘„Žƒ•–•Ǥ ›’‡”–‡•‹‘ ʹͲͲͺǢͷͳǣ͹ͲͶǦ͹ͳͳǤ 567 Cellular Physiology and Biochemistry 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 Cell Physiol Biochem 2014;33:557-568 DOI: 10.1159/000358634 Published online: February 25, 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion ‹†‘˜ƒǡŠ‡”‡’ƒ‘˜ƒǣš‹†‹œ‡†’Š‘•’Š‘Ž‹’‹†•‹†—…‡’Š‡‘–›’‹…•™‹–…Š‹‰‘ˆ˜ƒ•…—Žƒ”•‘‘–Š —•…Ž‡…‡ŽŽ•in vivo ƒ†in vitro.‹”…‡•ʹͲͲͻǢͳͲͶǣ͸ͲͻǦ͸ͳͺ ’Ž‹ǡŠ”‹•–‡•‡ ǡ…Š‡‹†‡”ǡ‡›†‘”ǡ ƒ‡Ž–‘ˆ–ǡŒ‘Ž„›‡ǣŠ‡ƒ‰‹‘–‡•‹–›’‡ ͳ”‡…‡’–‘”ƒ…–‹˜ƒ–‡•‡š–”ƒ…‡ŽŽ—Žƒ”•‹‰ƒŽǦ”‡‰—Žƒ–‡†‹ƒ•‡•ͳƒ†ʹ„› ’”‘–‡‹Ǧ†‡’‡†‡–ƒ†Ǧ ‹†‡’‡†‡–’ƒ–Š™ƒ›•‹…ƒ”†‹ƒ…›‘…›–‡•ƒ†Žƒ‰‡†‘”ˆˆǦ’‡”ˆ—•‡†Š‡ƒ”–•Ǥƒ•‹…Ž‹Šƒ”ƒ…‘Ž‘š‹…‘Ž ʹͲͲ͹ǢͳͲͲǣʹͺͻǦͻͷǤ — ǡ‹‰ ǡŠƒ‰ǡ‘‰ǡŠƒ‘ ǡƒ‰ǡŠ‘‰ǡ—ǡŠƒ‰ǣš‘‰‡‘—•Š›†”‘‰‡ •—Žϐ‹†‡’”‡˜‡–•…ƒ”†‹‘›‘…›–‡ƒ’‘’–‘•‹•ˆ”‘…ƒ”†‹ƒ…Š›’‡”–”‘’Š›‹†—…‡†„›‹•‘’”‘–‡”‡‘ŽǤ‘Ž‡ŽŽ ‹‘…Š‡ʹͲͳ͵Ǣ͵ͺͳǣͶͳȂͷͲ Š‘‰ǡƒ‰ǡƒ‰ǡ‡‰ǡ‘‰ǡŠƒ‘ ǡ‡ǡ‹ǡ—ǡ— ǡŠƒ‰ǣš‘‰‡‘—• Š›†”‘‰‡•—Žϐ‹†‡ƒ––‡—ƒ–‡•†‹ƒ„‡–‹…›‘…ƒ”†‹ƒŽ‹Œ—”›–Š”‘—‰Š…ƒ”†‹ƒ…‹–‘…Š‘†”‹ƒŽ’”‘–‡…–‹‘Ǥ‘Ž‡ŽŽ ‹‘…Š‡ʹͲͳʹǢ͵͹ͳǣͳͺ͹Ȃͳͻͺ Š‡‰ ǡ— ǣ‡…Šƒ‹…ƒŽ•–”‡–…Š•‹—Žƒ–‡•’”‘Ž‹ˆ‡”ƒ–‹‘‘ˆ˜‡‘—••‘‘–Š—•…Ž‡…‡ŽŽ•–Š”‘—‰Šƒ…–‹˜ƒ–‹‘ ‘ˆ–Š‡‹•—Ž‹ǦŽ‹‡‰”‘™–Šˆƒ…–‘”Ǧͳ”‡…‡’–‘”Ǥ”–‡”‹‘•…Ž‡”Š”‘„ƒ•…‹‘ŽʹͲͲ͹Ǣʹ͹ǣͳ͹ͶͶȂͳ͹ͷͳǤ Šƒ‹ƒ ǡ ƒ–ϐ‹‡Ž†ǡƒ”†ǡŠƒ‹ƒǣ˜‹†‡…‡ˆ‘”ƒˆ—…–‹‘ƒŽ…ƒŽ…‹—Ǧ•‡•‹‰”‡…‡’–‘” –Šƒ–‘†—Žƒ–‡•›‘‰‡‹…–‘‡‹”ƒ–•—„…—–ƒ‡‘—••ƒŽŽƒ”–‡”‹‡•Ǥ Š›•‹‘Ž‡ƒ”–‹”…Š›•‹‘Ž ʹͲͲͷǢʹͺͺǣͳ͹ͷ͸Ȃͳ͹͸ʹǤ ”‘™ǡ ƒ„ƒ ǡ‹……ƒ”†‹ǡ‘„ƒ”†‹ǡ—––‡”•ǡ‹ˆ‘”ǡ—ǡ‡†‹‰‡”ǡ›––‘ ǡ‡„‡”–ǣ Ž‘‹‰ƒ†…Šƒ”ƒ…–‡”‹œƒ–‹‘‘ˆƒ‡š–”ƒ…‡ŽŽ—Žƒ”ƒʹΪ•‡•‹‰”‡…‡’–‘”ˆ”‘„‘˜‹‡’ƒ”ƒ–Š›”‘‹†Ǥƒ–—”‡ ͳͻͻ͵Ǣ͵͸͸ǣͷ͹ͷȂͷͺͲǤ ‘‹‰”ƒ˜‡ǡ—‹ ǡ”‘™ǣǦ‹‘ƒ…‹†•‡•‹‰„›–Š‡‡š–”ƒ…‡ŽŽ—Žƒ”ƒʹΪǦ•‡•‹‰”‡…‡’–‘”Ǥ”‘… ƒ–Ž…ƒ†…‹ʹͲͲͲǢͻ͹ǣͶͺͳͶȂͶͺͳͻǤ ƒ‰ǡ—ǡŠƒ‘ǡŠƒ‰ ǡƒ‘ǡƒ‰ǡ—ǣƒŽ…‹—ƒ†’‘Ž›ƒ‹‡Ǧ”‡‰—Žƒ–‡†…ƒŽ…‹—Ǧ•‡•‹‰ ”‡…‡’–‘”•‹…ƒ”†‹ƒ…–‹••—‡•Ǥ—” ‹‘…Š‡ʹͲͲ͵Ǣʹ͹Ͳǣʹ͸ͺͲȂʹ͸ͺͺǤ — ǡ‹‡ ǡŠƒ‰ǡ•—Œ‹ƒ™ƒǡ —•…‘ǡ‹Ž˜‡”ƒǡ‹ƒ‰ǡ—‡ǡǣ͹Ǧ‡†‹ƒ–‡†ƒʹΪ•‹‰ƒŽ• …‘ˆ‡”ϐ‹„”‘‰‡‡•‹•‹Š—ƒ–”‹ƒŽ ‹„”‹ŽŽƒ–‹‘‹”…‡•ǤʹͲͳͲǢͳͲ͸ǣͻͻʹȂͳͲͲ͵ ‡””‹†‰‡ ǡ‘‘–ƒǡ‘†‡”‹…ǣƒŽ…‹—•‹‰ƒŽŽ‹‰ǣ†›ƒ‹…•ǡŠ‘‡‘•–ƒ•‹•ƒ†”‡‘†‡ŽŽ‹‰Ǥƒ– ‡˜‘Ž‡ŽŽ‹‘ŽʹͲͲ͵ǢͶǣͷͳ͹ȂͷʹͻǤ ‹ ǡ—ǡŠ‡ ǡŠ‘—ǡ•‡ ǡƒ—ǣŠƒ”ƒ…–‡”‹œƒ–‹‘‘ˆ—Ž–‹’Ž‡‹‘…Šƒ‡Ž‹…—Ž–—”‡†Š—ƒ …ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•Ǥ‘•‡ʹͲͲͻǢͳͲǢ͹͵Ͳ͹Ǧ͹͵ͳ͹ ‡‡ ǡ—Ǧǡ‡™‹•ǡ”‘—…Š ǡ—Ž˜‡”•–‘ǡƒ•‘ǡ‘‹‰”ƒ˜‡ǣŽŽ‘•–‡”‹…ƒ…–‹˜ƒ–‹‘‘ˆ –Š‡‡š–”ƒ…‡ŽŽ—Žƒ”ƒʹΪǦ•‡•‹‰”‡…‡’–‘”„›Ǧƒ‹‘ƒ…‹†•‡Šƒ…‡•ͳȀʹ’Š‘•’Š‘”›Žƒ–‹‘Ǥ‹‘…Š‡  ʹͲͲ͹ǢͶͲͶǣͳͶͳȂͳͶͻǤ ‘‘–ƒǡ‡””‹†‰‡ ǣŠ‡‡Ž‡‡–ƒŽ’”‹…‹’Ž‡•‘ˆ…ƒŽ…‹—•‹‰ƒŽ‹‰Ǥ‡ŽŽͳͻͻͷǢͺ͵ǣ͸͹ͷǦ͸͹ͺ ‡””‹†‰‡ ǡ‹’’ǡ‘‘–ƒǣŠ‡˜‡”•ƒ–‹Ž‹–›ƒ†—‹˜‡”•ƒŽ‹–›‘ˆ…ƒŽ…‹—•‹‰ƒŽ‹‰Ǥƒ–‡˜‘Ž‡ŽŽ ‹‘ŽʹͲͲͲǢͳǣͳͳǦʹͳ ™›‰Š‡†ƒ—™ǣ‘Ž‡…—Žƒ”‡…Šƒ‹••‘ˆ›‘…ƒ”†‹ƒŽ”‡‘†‡Ž‹‰ǤŠ›•‹‘Ž‡˜ͳͻͻͻǢ͹ͻǣʹͳͷȂʹ͸ʹǤ ƒƒ„‡ǡŠ‹†‘ǡƒ‰ƒ‹ǣ ‡‡š’”‡••‹‘‹ ‹„”‘„Žƒ•–•ƒ† ‹„”‘•‹•ǣ˜‘Ž˜‡‡–‹ƒ”†‹ƒ… ›’‡”–”‘’Š›Ǥ‹”…‡•ʹͲͲʹǢͻͳǣͳͳͲ͵Ȃͳͳͳ͵Ǥ ˜‡”‡––ǡ–ǢŽ‰‹ǡ ǣ–”‹ƒŽϐ‹„”‘•‹•ƒ†–Š‡‡…Šƒ‹••‘ˆƒ–”‹ƒŽϐ‹„”‹ŽŽƒ–‹‘Ǥ‡ƒ”–Š›–ŠʹͲͲ͹ǢͶǣʹͶȂ ʹ͹Ǥ Žƒ’Šƒǣ…Šƒ‡Ž•ƒ•…‡ŽŽ—Žƒ”•‡•‘”•Ǥƒ–—”‡ʹͲͲ͵ǢͶʹ͸ǣͷͳ͹ȂͷʹͶǤ ƒ…‡ƒǡ‘Žˆ‡ ǡŽƒ’Šƒǣ —…–‹‘ƒŽ͹Šƒ‡Ž•……——Žƒ–‡ƒ––Š‡Žƒ•ƒ‡„”ƒ‡‹ ‡•’‘•‡–‘ Ž—‹† Ž‘™Ǥ‹”…‡•ʹͲͲ͸ǢͻͺǣʹͶͷȂʹͷ͵Ǥ ƒ”–•ǡ‹Šƒ”ƒǡ‡‹ǡ‹‘‰ ǡ”—†‹‡ǡ‡”™‹•‹ǡƒ…‘ƒŽ† ǡ›‹ƒ•‹ǣ‡›”‘Ž‡ˆ‘” ͹…Šƒ‡Ž•‹ƒ‘š‹…‡—”‘ƒŽ†‡ƒ–ŠǤ‡ŽŽʹͲͲ͵Ǣͳͳͷǣͺ͸͵Ȃͺ͹͹Ǥ 568 Copyright: S Karger AG, Basel 2014 Reproduced with the permission of S Karger AG, Basel Further reproduction or distribution (electronic or otherwise) is prohibited without permission from the copyright holder  ... 2014 © 2014 S Karger AG, Basel www.karger.com/cpb Zhang et al.: CaR Regulating Cardiac Fibroblast Proliferation and Secretion Introduction ƒ”†‹ƒ…ϐ‹„”‘„Žƒ•–•ƒ”‡–Š‡ƒ‹‘Ǧ›‘…›–‡…‡ŽŽ–›’‡‹–Š‡Š‡ƒ”–Ǥ†‡”’Š›•‹‘Ž‘‰‹…ƒŽ... 2-yl)-2,5-diphenyltetrazolium bromide), cell count and the cell scratch assay ECM production was detected by expression of matrix metalloproteinase-3 and -9 (MMP-3 and -9) Activation RI &D5 SURPRWHG FDUGLDF... 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