Decreased uncoupling protein 2 expression in aging retinal pigment epithelial cells
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摘要
AIM: To analyze the expression of uncoupling protein 2(UCP2) in retinal pigment epithelium(RPE) cells at the different human age, further explore the possible new target of RPE cells protection.METHODS: Adult retinal pigment epithelial-19(ARPE-19) cells and the primary RPE cells at the different age(9-20 y,50-55 y, 60-70 y, >70 y) were cultured and harvested. The expression of UCP2 in these cells was detected by reverse transcription-polymerase chain reaction(RT-PCR), Western blot and confocal microscopy.RESULTS: Cells from the donors more than 60 y are larger and more fibroblastic in appearance compared to ARPE-19 cells and those primary cultures obtained from the younger individuals by using phase-contrast micrographs. Results of RT-PCR, Western blot and confocal microscopy all showed that UCP2 was highly expressed in ARPE-19 cells and in the younger primary cultured human RPE cells at the age of 9-20 y and 50-55 y, whereas lower expression of UCP2 was measured in the older primary cultured human RPE cells at the age more than 60 y.CONCLUSION: Expression of UCP2 gene is decreased in aged RPE cells, promoting the lower ability of anti-oxidation in these cells. It is indicated that UCP2 gene might be a new target for protecting the cells from oxidative stress damage.
AIM: To analyze the expression of uncoupling protein 2(UCP2) in retinal pigment epithelium(RPE) cells at the different human age, further explore the possible new target of RPE cells protection.METHODS: Adult retinal pigment epithelial-19(ARPE-19) cells and the primary RPE cells at the different age(9-20 y,50-55 y, 60-70 y, >70 y) were cultured and harvested. The expression of UCP2 in these cells was detected by reverse transcription-polymerase chain reaction(RT-PCR), Western blot and confocal microscopy.RESULTS: Cells from the donors more than 60 y are larger and more fibroblastic in appearance compared to ARPE-19 cells and those primary cultures obtained from the younger individuals by using phase-contrast micrographs. Results of RT-PCR, Western blot and confocal microscopy all showed that UCP2 was highly expressed in ARPE-19 cells and in the younger primary cultured human RPE cells at the age of 9-20 y and 50-55 y, whereas lower expression of UCP2 was measured in the older primary cultured human RPE cells at the age more than 60 y.CONCLUSION: Expression of UCP2 gene is decreased in aged RPE cells, promoting the lower ability of anti-oxidation in these cells. It is indicated that UCP2 gene might be a new target for protecting the cells from oxidative stress damage.
AIM: To analyze the expression of uncoupling protein 2(UCP2) in retinal pigment epithelium(RPE) cells at the different human age, further explore the possible new target of RPE cells protection.METHODS: Adult retinal pigment epithelial-19(ARPE-19) cells and the primary RPE cells at the different age(9-20 y,50-55 y, 60-70 y, >70 y) were cultured and harvested. The expression of UCP2 in these cells was detected by reverse transcription-polymerase chain reaction(RT-PCR), Western blot and confocal microscopy.RESULTS: Cells from the donors more than 60 y are larger and more fibroblastic in appearance compared to ARPE-19 cells and those primary cultures obtained from the younger individuals by using phase-contrast micrographs. Results of RT-PCR, Western blot and confocal microscopy all showed that UCP2 was highly expressed in ARPE-19 cells and in the younger primary cultured human RPE cells at the age of 9-20 y and 50-55 y, whereas lower expression of UCP2 was measured in the older primary cultured human RPE cells at the age more than 60 y.CONCLUSION: Expression of UCP2 gene is decreased in aged RPE cells, promoting the lower ability of anti-oxidation in these cells. It is indicated that UCP2 gene might be a new target for protecting the cells from oxidative stress damage.
引文
1 D’Cruz PM,Yasumura D,Weir J,Matthes MT,Abderrahim H,LaVail MM,Vollrath D.Mutation of the receptor tyrosine kinase gene Mertk in the retinal dystrophic RCS rat.Hum Mol Genet 2000;9(4):645-651.
2 Gal A,Li Y,Thompson DA,Weir J,Orth U,Jacobson SG,ApfelstedtSylla E,Vollrath D.Mutations in MERTK,the human orthologue of the RCS rat retinal dystrophy gene,cause retinitis pigmentosa.Nat Genet2000;26(3):270-271.
3 Dorey CK,Wu G,Ebenstein D,Garsd A,Weiter JJ.Cell loss in the aging retina.Relationship to lipofuscin accumulation and macular degeneration.Invest Ophthalmol Vis Sci 1989;30(8):1691-1699.
4 Green WR,Enger C.Age-related macular degeneration histopathologic studies.The 1992 Lorenz E.Zimmerman Lecture.Ophthalmology1993;100(10):1519-1535.
5 Beatty S,Koh H,Phil M,Henson D,Boulton M.The role of oxidative stress in the pathogenesis of age-related macular degeneration.Surv Ophthalmol 2000;45(2):115-134.
6 Dunaief JL,Dentchev T,Ying GS,Milam AH.The role of apoptosis in age-related macular degeneration.Arch Ophthalmol 2002;120(11):1435-1442.
7 Winkler BS,Boulton ME,Gottsch JD,Sternberg P.Oxidative damage and age-related macular degeneration.Molr Vis 1999;5:32.
8 Shen C,Ma W,Zheng W,Huang H,Xia R,Li C,Zhu X.The antioxidant effects of riluzole on the APRE-19 celll model injury-induced by t-BHP.BMC Ophthalmol 2017;17(1):210.
9 Sachdeva MM,Cano M,Handa JT.Nrf2 signaling is impaired in the aging RPE given an oxidative insult.Exp Eye Res 2014;119:111-114.
10 Iacovelli J,Rowe GC,Khadka A,Diaz-Aguilar D,Spencer C,Arany Z,Saint-Geniez M.PGC-1αinduces human RPE oxidative metabolism and antioxidant capacity.Invest Ophthalmol Vis Sci 2016;57(3):1038.
11 Wang J,Gong HM,Zou HH,Liang L,Wu XY.Isorhamnetin prevents H2O2-induced oxidative stress in human retinal pigment epithelial cells.Mol Med Rep 2018;17(1):648-652.
12 Hong Y,Fink BD,Dillon JS,Sivitz WI.Effects of adenoviral overexpression of uncoupling protein-2 and-3 on mitochondrial respiration in insulinoma cells.Endocrinology 2001;142(1):249-256.
13 Wang X,Axelsson J,Nordfors L,Qureshi AR,Avesani C,Barany P,Schalling M,Heimbürger O,Lindholm B,Stenvinkel P.Changes in fat mass after initiation of maintenance dialysis is influenced by the uncoupling protein 2 exon 8 insertion/deletion polymorphism.Nephrol Dial Transplant 2007;22(1):196-202.
14 Horimoto M,Resnick MB,Konkin TA,Routhier J,Wands JR,Baffy G.Expression of uncoupling protein-2 in human colon cancer.Clin Cancer Res 2004;10(18 Pt1):6203-6207.
15 He Y,Leung KW,Ren Y,Pei JZ,Ge J,Tombran-Tink J.PEDFimproves mitochondrial function in RPE cells during oxidative stress.Invest Ophthalmol Vis Sci 2014;55(10):6742-6755.
16 Dong X,Li ZR,Wang W,Zhang WJ,Liu SZ,Zhang XM,Fang J,Maeda H,Matsukura M.Protective effect of canolol from oxidative stressinduced cell damage in ARPE-19 cells via an ERK mediated antioxidative pathway.Mol Vis 2011;17:2040-2048.
17 Lin H,Qian J,Castillo AC,Long B,Keyes KT,Chen G,Ye Y.Effect of miR-23 on oxidant-induced injury in human retinal pigment epithelial cells.Invest Ophthalmol Vis Sci 2011;52(9):6308-6314.
18 Zhuge CC,Xu JY,Zhang J,Li W,Li P,Li Z,Chen L,Liu X,Shang P,Xu H,Lu Y,Wang F,Lu L,Xu GT.Fullerenol protects retinal pigment epithelial cells from oxidative stress-induced premature senescence via activating SIRT1.Invest Ophthalmol Vis Sci 2014;55(7):4628-4638.
19 Cruz-Guilloty F,Saeed AM,Duffort S,Cano M,Ebrahimi KB,Ballmick A,Tan YH,Wang H,Laird JM,Salomon RG,Handa JT,Perez VL.T cells and macrophages responding to oxidative damage cooperate in pathogenesis of a mouse model of age-related macular degeneration.PLo S One 2014;9(2):e88201.
20 Promsote W,Veeranan-Karmegam R,Ananth S,Shen DF,Chan CC,Lambert NA,Ganapathy V,Martin PM.L-2-oxothiazolidine-4-carboxylic acid attenuates oxidative stress and inflammation in retinal pigment epithelium.Mol Vis 2014;20:73-88.
21 Chen C,Cano M,Wang JJ,Li J,Huang C,Yu Q,Herbert TP,Handa JT,Zhang SX.Role of unfolded protein response dysregulation in oxidative injury of retinal pigment epithelial cells.Antioxid Redox Signal2014;20(14):2091-2106.
22 Sheu SJ,Liu NC,Ou CC,Bee YS,Chen SC,Lin HC,Chan JY.Resveratrol stimulates mitochondrial bioenergetics to protect retinal pigment epithelial cells from oxidative damage.Invest Ophthalmol Vis Sci2013;54(9):6426-6438.
23 Plafker SM,O’Mealey GB,Szweda LI.Mechanisms for countering oxidative stress and damage in retinal pigment epithelium.Int Rev Cell Mol Biol 2012;298:135-177.
24 Jitsanong T,Khanobdee K,Piyachaturawat P,Wongprasert K.Diarylheptanoid 7-(3,4 dihydroxyphenyl)-5-hydroxy-1-phenyl-(1E)-1-heptene from Curcuma comosa Roxb.protects retinal pigment epithelial cells against oxidative stress-induced cell death.Toxicol In Vitro2011;25(1):167-176.
25 Faghiri Z,Bazan NG.PI3K/Akt and mTOR/p70S6K pathways mediate neuroprotectin D1-induced retinal pigment epithelial cell survival during oxidative stress-induced apoptosis.Exp Eye Res 2010;90(6):718-725.
26 Wang ZY,Shen LJ,Tu L,Hu DN,Liu GY,Zhou ZL,Lin Y,Chen LH,Qu J.Erythropoietin protects retinal pigment epithelial cells from oxidative damage.Free Radic Biol Med 2009;46(8):1032-1041.
27 Davis MD,Gangnon RE,Lee LY,Hubbard LD,Klein BE,Klein R,Ferris FL,Bressler SB,Milton RC,Age-Related Eye Disease Study Group.The age-related eye disease study severity scale for agerelated macular degeneration:AREDS report no.17.Arch Ophthalmol2005;123(11):1484-1498.
28 Doughan AK,Harrison DG,Dikalov SI.Molecular mechanisms of angiotensin II-mediated mitochondrial dysfunction:linking mitochondrialoxidative damage and vascular endothelial dysfunction.Circ Res2008;102(4):488-496.
29 Noh YH,Kim KY,Shim MS,Choi SH,Choi S,Ellisman MH,Weinreb RN,Perkins GA,Ju WK.Inhibition of oxidative stress by coenzyme Q10increases mitochondrial mass and improves bioenergetic function in optic nerve head astrocytes.Cell Death Dis 2013;4:e820.
30 Di Castro S,Scarpino S,Marchitti S,Bianchi F,Stanzione R,Cotugno M,Sironi L,Gelosa P,Duranti E,Ruco L,Volpe M,Rubattu S.Differential modulation of uncoupling protein 2 in kidneys of strokeprone spontaneously hypertensive rats under high-salt/low-potassium diet.Hypertension 2013;61(2):534-541.
31 Fu ZH,Zhou YH,Zhu WJ,Chen XM,Li XM,Tan Z.Uncoupling protein 2 combats oxidative damage to human sperm.Zhonghua Nan Ke Xue 2010;16(6):516-519.
32 Müller S,Kaiser H,Krüger B,Fitzner B,Lange F,Bock CN,Nizze H,Ibrahim SM,Fuellen G,Wolkenhauer O,Jaster R.Age-dependent effects of UCP2 deficiency on experimental acute pancreatitis in mice.PLo S One2014;9(4):e94494.
33 Harper ME,Bevilacqua L,Hagopian K,Weindruch R,Ramsey JJ.Ageing,oxidative stress,and mitochondrial uncoupling.Acta Physiol Scand 2004;182(4):321-331.
34 Dietrich MO,Horvath TL.The role of mitochondrial uncoupling proteins in lifespan.Pflugers Arch 2010;459(2):269-275.
35 Andrews ZB.Uncoupling protein-2 and the potential link between metabolism and longevity.Curr Aging Sci 2010;3(2):102-112.
2 Gal A,Li Y,Thompson DA,Weir J,Orth U,Jacobson SG,ApfelstedtSylla E,Vollrath D.Mutations in MERTK,the human orthologue of the RCS rat retinal dystrophy gene,cause retinitis pigmentosa.Nat Genet2000;26(3):270-271.
3 Dorey CK,Wu G,Ebenstein D,Garsd A,Weiter JJ.Cell loss in the aging retina.Relationship to lipofuscin accumulation and macular degeneration.Invest Ophthalmol Vis Sci 1989;30(8):1691-1699.
4 Green WR,Enger C.Age-related macular degeneration histopathologic studies.The 1992 Lorenz E.Zimmerman Lecture.Ophthalmology1993;100(10):1519-1535.
5 Beatty S,Koh H,Phil M,Henson D,Boulton M.The role of oxidative stress in the pathogenesis of age-related macular degeneration.Surv Ophthalmol 2000;45(2):115-134.
6 Dunaief JL,Dentchev T,Ying GS,Milam AH.The role of apoptosis in age-related macular degeneration.Arch Ophthalmol 2002;120(11):1435-1442.
7 Winkler BS,Boulton ME,Gottsch JD,Sternberg P.Oxidative damage and age-related macular degeneration.Molr Vis 1999;5:32.
8 Shen C,Ma W,Zheng W,Huang H,Xia R,Li C,Zhu X.The antioxidant effects of riluzole on the APRE-19 celll model injury-induced by t-BHP.BMC Ophthalmol 2017;17(1):210.
9 Sachdeva MM,Cano M,Handa JT.Nrf2 signaling is impaired in the aging RPE given an oxidative insult.Exp Eye Res 2014;119:111-114.
10 Iacovelli J,Rowe GC,Khadka A,Diaz-Aguilar D,Spencer C,Arany Z,Saint-Geniez M.PGC-1αinduces human RPE oxidative metabolism and antioxidant capacity.Invest Ophthalmol Vis Sci 2016;57(3):1038.
11 Wang J,Gong HM,Zou HH,Liang L,Wu XY.Isorhamnetin prevents H2O2-induced oxidative stress in human retinal pigment epithelial cells.Mol Med Rep 2018;17(1):648-652.
12 Hong Y,Fink BD,Dillon JS,Sivitz WI.Effects of adenoviral overexpression of uncoupling protein-2 and-3 on mitochondrial respiration in insulinoma cells.Endocrinology 2001;142(1):249-256.
13 Wang X,Axelsson J,Nordfors L,Qureshi AR,Avesani C,Barany P,Schalling M,Heimbürger O,Lindholm B,Stenvinkel P.Changes in fat mass after initiation of maintenance dialysis is influenced by the uncoupling protein 2 exon 8 insertion/deletion polymorphism.Nephrol Dial Transplant 2007;22(1):196-202.
14 Horimoto M,Resnick MB,Konkin TA,Routhier J,Wands JR,Baffy G.Expression of uncoupling protein-2 in human colon cancer.Clin Cancer Res 2004;10(18 Pt1):6203-6207.
15 He Y,Leung KW,Ren Y,Pei JZ,Ge J,Tombran-Tink J.PEDFimproves mitochondrial function in RPE cells during oxidative stress.Invest Ophthalmol Vis Sci 2014;55(10):6742-6755.
16 Dong X,Li ZR,Wang W,Zhang WJ,Liu SZ,Zhang XM,Fang J,Maeda H,Matsukura M.Protective effect of canolol from oxidative stressinduced cell damage in ARPE-19 cells via an ERK mediated antioxidative pathway.Mol Vis 2011;17:2040-2048.
17 Lin H,Qian J,Castillo AC,Long B,Keyes KT,Chen G,Ye Y.Effect of miR-23 on oxidant-induced injury in human retinal pigment epithelial cells.Invest Ophthalmol Vis Sci 2011;52(9):6308-6314.
18 Zhuge CC,Xu JY,Zhang J,Li W,Li P,Li Z,Chen L,Liu X,Shang P,Xu H,Lu Y,Wang F,Lu L,Xu GT.Fullerenol protects retinal pigment epithelial cells from oxidative stress-induced premature senescence via activating SIRT1.Invest Ophthalmol Vis Sci 2014;55(7):4628-4638.
19 Cruz-Guilloty F,Saeed AM,Duffort S,Cano M,Ebrahimi KB,Ballmick A,Tan YH,Wang H,Laird JM,Salomon RG,Handa JT,Perez VL.T cells and macrophages responding to oxidative damage cooperate in pathogenesis of a mouse model of age-related macular degeneration.PLo S One 2014;9(2):e88201.
20 Promsote W,Veeranan-Karmegam R,Ananth S,Shen DF,Chan CC,Lambert NA,Ganapathy V,Martin PM.L-2-oxothiazolidine-4-carboxylic acid attenuates oxidative stress and inflammation in retinal pigment epithelium.Mol Vis 2014;20:73-88.
21 Chen C,Cano M,Wang JJ,Li J,Huang C,Yu Q,Herbert TP,Handa JT,Zhang SX.Role of unfolded protein response dysregulation in oxidative injury of retinal pigment epithelial cells.Antioxid Redox Signal2014;20(14):2091-2106.
22 Sheu SJ,Liu NC,Ou CC,Bee YS,Chen SC,Lin HC,Chan JY.Resveratrol stimulates mitochondrial bioenergetics to protect retinal pigment epithelial cells from oxidative damage.Invest Ophthalmol Vis Sci2013;54(9):6426-6438.
23 Plafker SM,O’Mealey GB,Szweda LI.Mechanisms for countering oxidative stress and damage in retinal pigment epithelium.Int Rev Cell Mol Biol 2012;298:135-177.
24 Jitsanong T,Khanobdee K,Piyachaturawat P,Wongprasert K.Diarylheptanoid 7-(3,4 dihydroxyphenyl)-5-hydroxy-1-phenyl-(1E)-1-heptene from Curcuma comosa Roxb.protects retinal pigment epithelial cells against oxidative stress-induced cell death.Toxicol In Vitro2011;25(1):167-176.
25 Faghiri Z,Bazan NG.PI3K/Akt and mTOR/p70S6K pathways mediate neuroprotectin D1-induced retinal pigment epithelial cell survival during oxidative stress-induced apoptosis.Exp Eye Res 2010;90(6):718-725.
26 Wang ZY,Shen LJ,Tu L,Hu DN,Liu GY,Zhou ZL,Lin Y,Chen LH,Qu J.Erythropoietin protects retinal pigment epithelial cells from oxidative damage.Free Radic Biol Med 2009;46(8):1032-1041.
27 Davis MD,Gangnon RE,Lee LY,Hubbard LD,Klein BE,Klein R,Ferris FL,Bressler SB,Milton RC,Age-Related Eye Disease Study Group.The age-related eye disease study severity scale for agerelated macular degeneration:AREDS report no.17.Arch Ophthalmol2005;123(11):1484-1498.
28 Doughan AK,Harrison DG,Dikalov SI.Molecular mechanisms of angiotensin II-mediated mitochondrial dysfunction:linking mitochondrialoxidative damage and vascular endothelial dysfunction.Circ Res2008;102(4):488-496.
29 Noh YH,Kim KY,Shim MS,Choi SH,Choi S,Ellisman MH,Weinreb RN,Perkins GA,Ju WK.Inhibition of oxidative stress by coenzyme Q10increases mitochondrial mass and improves bioenergetic function in optic nerve head astrocytes.Cell Death Dis 2013;4:e820.
30 Di Castro S,Scarpino S,Marchitti S,Bianchi F,Stanzione R,Cotugno M,Sironi L,Gelosa P,Duranti E,Ruco L,Volpe M,Rubattu S.Differential modulation of uncoupling protein 2 in kidneys of strokeprone spontaneously hypertensive rats under high-salt/low-potassium diet.Hypertension 2013;61(2):534-541.
31 Fu ZH,Zhou YH,Zhu WJ,Chen XM,Li XM,Tan Z.Uncoupling protein 2 combats oxidative damage to human sperm.Zhonghua Nan Ke Xue 2010;16(6):516-519.
32 Müller S,Kaiser H,Krüger B,Fitzner B,Lange F,Bock CN,Nizze H,Ibrahim SM,Fuellen G,Wolkenhauer O,Jaster R.Age-dependent effects of UCP2 deficiency on experimental acute pancreatitis in mice.PLo S One2014;9(4):e94494.
33 Harper ME,Bevilacqua L,Hagopian K,Weindruch R,Ramsey JJ.Ageing,oxidative stress,and mitochondrial uncoupling.Acta Physiol Scand 2004;182(4):321-331.
34 Dietrich MO,Horvath TL.The role of mitochondrial uncoupling proteins in lifespan.Pflugers Arch 2010;459(2):269-275.
35 Andrews ZB.Uncoupling protein-2 and the potential link between metabolism and longevity.Curr Aging Sci 2010;3(2):102-112.