PPAR-γ受体激动剂罗格列酮对兔准分子激光角膜切削术后角膜上皮下雾状混浊的影响
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摘要
目的探讨过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor gamma,PPARγ)激动剂罗格列酮对兔准分子激光角膜切削术(photorefractive keratectomy,PRK)术后角膜上皮下雾状混浊(haze)的影响。方法选取普通级新西兰大白兔64只(64眼),随机分为28μmol·L~(-1)罗格列酮组、14μmol·L~(-1)罗格列酮组、1 g·L~(-1)二甲基亚砜(dimethyl sulfoxide,DMSO)+生理盐水组、单纯PRK组,每组16只(16眼)。均行右眼PRK手术,术后分别给予28μmol·L~(-1)罗格列酮、14μmol·L~(-1)罗格列酮、1 g·L~(-1)DMSO+生理盐水滴眼,单纯PRK组未作任何特殊处理。术后各组用裂隙灯观察并比较角膜上皮愈合情况;分别于PRK术后7 d、28 d比较各组haze情况并行眼前节照相;每组分别于PRK术后7 d、28 d分批处死8只兔,取角膜行免疫组织化学法比较各组转化生长因子-β1(transforming growth factor-β1,TGF-β1)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、基质金属蛋白酶-2(matrix metalloproteinases-2,MMP-2)的表达。结果 PRK术后角膜上皮愈合时间组间差异无统计学意义(P>0.05);PRK术后7 d、28 d,各组haze分级及TGF-β1、α-SMA、MMP-2的表达情况,组间差异均有统计学意义(均为P<0.05),其中28μmol·L~(-1)罗格列酮组haze最轻,14μmol·L~(-1)罗格列酮组其次,明显轻于1 g·L~(-1) DMSO+生理盐水组及单纯PRK组,除1 g·L~(-1)DMSO+生理盐水组及单纯PRK组组间差异均无统计学意义(均为P>0.05)外,其余组间两两比较差异均有统计学意义(均为P<0.05)。结论 PPAR-γ受体激动剂罗格列酮对兔PRK术后haze有抑制作用,呈浓度依赖性,可能由TGF-β1/Smad通路介导。
Objective To investigate the effects of peroxisome proliferator-activated receptor gamma(PPAR-γ) agonist rosiglitazone on corneal haze after photorefractive keratectomy(PRK) in rabbits.Methods Totally 64 New Zealand white rabbits were selected and randomly divided into four groups,28 μmol·L~(-1) rosiglitazone group,14 μmol·L~(-1) rosiglitazone group,1 g·L~(-1) dimethyl sulfoxide(DMSO)+normal saline(NS) group and PRK group,sixteen rabbits(16 eyes) in each group.PRK was performed on the right eye in all groups.28 μmol·L~(-1) rosiglitazone group,14 μmol·L~(-1) rosiglitazone group and 1 g·L~(-1) DMSO+NS group was given 28 μmol·L~(-1) rosiglitazone,14 μmol·L~(-1) rosiglitazone and 1 g·L~(-1) DMSO+NS eye drops,respectively,no intervention was performed in the PRK group.Slit lamp was used to observe and compare the corneal epithelium healing in each group.On the 7 th day and 28 th day after PRK,haze was compared and recorded by the anterior segment photographs.Eight rabbits were sacrificed in each group,all of the corneas were obtained and then immunohistochemistry was performed to detect the expressions of transforming growth factors beta 1(TGF-β1),alpha smooth muscle actin(α-SMA) and matrix metalloproteinase-2(MMP-2).Results There was no significant difference in healing time of corneal epithelium after PRK in each group(P>0.05).On the 7 th day and 28 th day after PRK,there were statistically significant differences in haze grade and the expressions of TGF-β1,α-SMA and MMP-2 in each group(both P<0.05).Haze classification and expressions of TGF-β1,α-SMA and MMP-2 in 28 μmol·L~(-1) rosiglitazone group were both the most light,followed by 14 μmol·L~(-1) rosiglitazone group,which significantly lighter than that in 1 g·L~(-1) DMSO+NS group and PRK group.Except that there was no statistical difference between 1 g·L~(-1) DMSO+NS group and PRK group(all P>0.05),and pairwise comparison was statistically significant between the other groups(all P<0.05).Conclusion PPAR-γ agonist rosiglitazone can inhibit corneal haze after PRK in rabbits,the effect of high concentration is more obvious than that of low concentration,which maybe mediated by the TGF-β1/Smad pathway.
Objective To investigate the effects of peroxisome proliferator-activated receptor gamma(PPAR-γ) agonist rosiglitazone on corneal haze after photorefractive keratectomy(PRK) in rabbits.Methods Totally 64 New Zealand white rabbits were selected and randomly divided into four groups,28 μmol·L~(-1) rosiglitazone group,14 μmol·L~(-1) rosiglitazone group,1 g·L~(-1) dimethyl sulfoxide(DMSO)+normal saline(NS) group and PRK group,sixteen rabbits(16 eyes) in each group.PRK was performed on the right eye in all groups.28 μmol·L~(-1) rosiglitazone group,14 μmol·L~(-1) rosiglitazone group and 1 g·L~(-1) DMSO+NS group was given 28 μmol·L~(-1) rosiglitazone,14 μmol·L~(-1) rosiglitazone and 1 g·L~(-1) DMSO+NS eye drops,respectively,no intervention was performed in the PRK group.Slit lamp was used to observe and compare the corneal epithelium healing in each group.On the 7 th day and 28 th day after PRK,haze was compared and recorded by the anterior segment photographs.Eight rabbits were sacrificed in each group,all of the corneas were obtained and then immunohistochemistry was performed to detect the expressions of transforming growth factors beta 1(TGF-β1),alpha smooth muscle actin(α-SMA) and matrix metalloproteinase-2(MMP-2).Results There was no significant difference in healing time of corneal epithelium after PRK in each group(P>0.05).On the 7 th day and 28 th day after PRK,there were statistically significant differences in haze grade and the expressions of TGF-β1,α-SMA and MMP-2 in each group(both P<0.05).Haze classification and expressions of TGF-β1,α-SMA and MMP-2 in 28 μmol·L~(-1) rosiglitazone group were both the most light,followed by 14 μmol·L~(-1) rosiglitazone group,which significantly lighter than that in 1 g·L~(-1) DMSO+NS group and PRK group.Except that there was no statistical difference between 1 g·L~(-1) DMSO+NS group and PRK group(all P>0.05),and pairwise comparison was statistically significant between the other groups(all P<0.05).Conclusion PPAR-γ agonist rosiglitazone can inhibit corneal haze after PRK in rabbits,the effect of high concentration is more obvious than that of low concentration,which maybe mediated by the TGF-β1/Smad pathway.
引文
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[5] CHOU H C,WEN L L,CHANG C C,LIN C Y,JIN L,JUAN S H.From the cover:l-Carnitine via pparγ- and sirt1-dependent mechanisms attenuates epithelial-mesenchymal transition and renal fibrosis caused by Perfluorooctanesulfonate[J].Toxicol Sci,2017,160(2):217-229.
[6] LATELLA G,VETUSCHI A,SFERRA R,SPECA S,GAUDIO E.Localization of ανβ6 integrin-TGF-β1/Smad3,mTOR and PPARγ in experimental colorectal fibrosis[J].Eur J Histochem,2013,57(4):e40.
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[11] SINGH V,TORRICELLI A A,NAYEB-HASHEMI N,AGRAWAL V,WILSON S E.Mouse strain variation in SMA(+) myofibroblast development after corneal injury[J].Exp Eye Res,2013,115(10):27.
[12] TOMáSJUAN J,MURUETAGOYENA L A,HANNEKEN L.Corneal regeneration after photorefractive keratectomy:a review[J].J Optom,2015,8(3):149.
[13] SAIKA S,YAMANAKA O,OKADA Y,MIYAMOTO T,KITANO A,FLANDERS K C,et al.Effect of overexpression of PPARgamma on the healing process of corneal alkali burn in mice[J].Am J Physiol Cell Physiol,2007,293(1):C75-86.
[14] YAMANAKA O,MIYAZAKI K,KITANO A,SAIKA S,NAKAJIMA Y,IKEDA K.Suppression of injury-induced conjunctiva scarring by peroxisome proliferator-activated receptor gamma gene transfer in mice[J].Invest Ophthalmol Vis Sci,2009,50(1):187-193.
[15] BI X D.PPARy agnoists inhibit cell growth and TGF-β2 induced extracellular matrix production in human Tenon’s fibroblasts[D].Shanghai:The Secnod Military Medical University,2011.毕晓达.PPARγ激动剂抑制人Tenon’s囊成纤维细胞增殖及TGF-β2诱导细胞外基质合成的研究[D].上海:第二军医大学,2011.
[16] HUXLIN K R,HINDMAN H B,JEON K I,BüHREN J,MACRAE S,DEMAGISTRIS M,et al.Topical rosiglitazone is an effective anti-scarring agent in the cornea[J].PLoS One,2013,8(8):e70785.
[17] GU L,ZHU Y J,YANG X,GUO Z J,XU W B,TIAN X L.Effect of TGF-beta/Smad signalingpathway on lung myofibroblast differentiation[J].Acta Pharmacol Sin,2007,28 (3):382-391.
[18] JAVELAUD D,MAUVIEL A.Transforming growth factor-betas:Smad signaling and roles in physiopathology[J].Pathol Biol,2004,52(1):50-54.
[19] VO N.CREB-binding protein and p300 in transcriptional regulation[J].J Biol Chem,2001,276(17):13505-13508.
[20] INOUE Y,ITOH Y,ABE K,OKAMOTO T,DAITOKU H,FUKAMIZU A,et al.Smad3 is acetylated by p300/CBP to regulate its transactivation activity[J].Oncogene,2007,26(4):500-508.
[21] GHOSH A K,BHATTACHARYYA S,WEI J,KIM S,BARAK Y,MORI Y,et al.Peroxisome proliferator-activated receptor-gamma abrogates Smad-dependent collagen stimulation by targeting the p300 transcriptional coactivator[J].FASEB J,2009,23(9):2968-2977.
[22] GHOSH A K.The transcriptional coactivator and acetyltransferase p300 in fibroblast biology and fibrosis[J].J Cell Physiol,2007,213(3):663-671.
[23] LIM J Y,OH M A,KIM W H,SOHN H Y,PARK S I.AMP-activated protein kinase inhibits TGF-β-induced fibrogenic responses of hepatic stellate cells by targeting transcriptional coactivator p300[J].J Cell Physiol,2012,227(3):1081-1089.
[24] ZHENG X,QI C,ZHANG S,FANG Y,NING W.TGF-β1 induces Fstl1 via the Smad3-c-Jun pathway in lung fibroblasts[J].Am J Physiol Lung Cell Mol Physiol,2017,313(2):L240-251.
[2] NETTO M V,MOHAN R R,AMBRóSIO R J R,HUTCHEON A E,ZIESKE J D,WILSON S E.Wound healing in the cornea:a review of refractive surgery complications and new prospects for therapy[J].Cornea,2005,24(5):509-522.
[3] JESTER J V,NIEN C J,VASILIOU V.Quiescent keratocytes fail to repair MMC induced DNA damage leading to the long-term inhibition of myofibroblast differentiation and wound healing[J].Mol Vis,2012,18:1828-1839.
[4] DELRIO C,CANTARERO I,PALOMARES B,GóMEZ-CA?AS M,FERNáNDEZ-RUIZ J,PAVICIC C,et al.VCE-0043,a cannabidiol aminoquinone derivative,prevents bleomycin-induced skin fibrosis and inflammation through PPARγ- and CB receptor-dependent pathways[J].Br J Pharmacol,2018,175(19):3813-3831.
[5] CHOU H C,WEN L L,CHANG C C,LIN C Y,JIN L,JUAN S H.From the cover:l-Carnitine via pparγ- and sirt1-dependent mechanisms attenuates epithelial-mesenchymal transition and renal fibrosis caused by Perfluorooctanesulfonate[J].Toxicol Sci,2017,160(2):217-229.
[6] LATELLA G,VETUSCHI A,SFERRA R,SPECA S,GAUDIO E.Localization of ανβ6 integrin-TGF-β1/Smad3,mTOR and PPARγ in experimental colorectal fibrosis[J].Eur J Histochem,2013,57(4):e40.
[7] FANTES F E,HANNA K D,WARING GO R D,POULIQUEN Y,THOMPSON K P,SAVOLDELLI M.Wound healing after excimer laser keratomileusis (photorefractive keratectomy) in monkeys[J].Arch Ophthalmol,1990,108(5):665-675.
[8] LALOCK T D,GIBSON D J,DUNCAN M R,TULI S S,GROTENDORST G R,SCHULTZ G S.A connective tissue growth factor signaling receptor in corneal fibroblasts[J].Invest Ophthalmol Vis Sci,2012,53(7):3387.
[9] IBSON D J,PI L,SRIRAM S,MAO C,PETERSEN B E,SCOTT E W,et al.Conditional knockout of CTGF affects corneal wound healing[J].Invest Ophthalmol Vis Sci,2014,55(4):2062-2070.
[10] KARAMICHOS D,GUO X Q,HUTCHEON A E,ZIESKE J D.Human corneal fibrosis:an in vitro model[J].Invest Ophthalmol Vis Sci,2010,51(3):1382.
[11] SINGH V,TORRICELLI A A,NAYEB-HASHEMI N,AGRAWAL V,WILSON S E.Mouse strain variation in SMA(+) myofibroblast development after corneal injury[J].Exp Eye Res,2013,115(10):27.
[12] TOMáSJUAN J,MURUETAGOYENA L A,HANNEKEN L.Corneal regeneration after photorefractive keratectomy:a review[J].J Optom,2015,8(3):149.
[13] SAIKA S,YAMANAKA O,OKADA Y,MIYAMOTO T,KITANO A,FLANDERS K C,et al.Effect of overexpression of PPARgamma on the healing process of corneal alkali burn in mice[J].Am J Physiol Cell Physiol,2007,293(1):C75-86.
[14] YAMANAKA O,MIYAZAKI K,KITANO A,SAIKA S,NAKAJIMA Y,IKEDA K.Suppression of injury-induced conjunctiva scarring by peroxisome proliferator-activated receptor gamma gene transfer in mice[J].Invest Ophthalmol Vis Sci,2009,50(1):187-193.
[15] BI X D.PPARy agnoists inhibit cell growth and TGF-β2 induced extracellular matrix production in human Tenon’s fibroblasts[D].Shanghai:The Secnod Military Medical University,2011.毕晓达.PPARγ激动剂抑制人Tenon’s囊成纤维细胞增殖及TGF-β2诱导细胞外基质合成的研究[D].上海:第二军医大学,2011.
[16] HUXLIN K R,HINDMAN H B,JEON K I,BüHREN J,MACRAE S,DEMAGISTRIS M,et al.Topical rosiglitazone is an effective anti-scarring agent in the cornea[J].PLoS One,2013,8(8):e70785.
[17] GU L,ZHU Y J,YANG X,GUO Z J,XU W B,TIAN X L.Effect of TGF-beta/Smad signalingpathway on lung myofibroblast differentiation[J].Acta Pharmacol Sin,2007,28 (3):382-391.
[18] JAVELAUD D,MAUVIEL A.Transforming growth factor-betas:Smad signaling and roles in physiopathology[J].Pathol Biol,2004,52(1):50-54.
[19] VO N.CREB-binding protein and p300 in transcriptional regulation[J].J Biol Chem,2001,276(17):13505-13508.
[20] INOUE Y,ITOH Y,ABE K,OKAMOTO T,DAITOKU H,FUKAMIZU A,et al.Smad3 is acetylated by p300/CBP to regulate its transactivation activity[J].Oncogene,2007,26(4):500-508.
[21] GHOSH A K,BHATTACHARYYA S,WEI J,KIM S,BARAK Y,MORI Y,et al.Peroxisome proliferator-activated receptor-gamma abrogates Smad-dependent collagen stimulation by targeting the p300 transcriptional coactivator[J].FASEB J,2009,23(9):2968-2977.
[22] GHOSH A K.The transcriptional coactivator and acetyltransferase p300 in fibroblast biology and fibrosis[J].J Cell Physiol,2007,213(3):663-671.
[23] LIM J Y,OH M A,KIM W H,SOHN H Y,PARK S I.AMP-activated protein kinase inhibits TGF-β-induced fibrogenic responses of hepatic stellate cells by targeting transcriptional coactivator p300[J].J Cell Physiol,2012,227(3):1081-1089.
[24] ZHENG X,QI C,ZHANG S,FANG Y,NING W.TGF-β1 induces Fstl1 via the Smad3-c-Jun pathway in lung fibroblasts[J].Am J Physiol Lung Cell Mol Physiol,2017,313(2):L240-251.