[1]王思文,李鹏飞,王从玉,等.褪黑素对过氧化氢诱导的人晶状体上皮细胞凋亡及自噬的影响[J].眼科新进展,2023,43(11):848-852.[doi:10.13389/j.cnki.rao.2023.0170]
　WANG Siwen,LI Pengfei,WANG Congyu,et al.Effect of melatonin on hydrogen peroxide-induced apoptosis and autophagy in human lens epithelial cells[J].Recent Advances in Ophthalmology,2023,43(11):848-852.[doi:10.13389/j.cnki.rao.2023.0170]
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褪黑素对过氧化氢诱导的人晶状体上皮细胞凋亡及自噬的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
43卷
期数:
2023年11期
页码:
848-852
栏目:
实验研究
出版日期:
2023-11-05

文章信息/Info

Title:
Effect of melatonin on hydrogen peroxide-induced apoptosis and autophagy in human lens epithelial cells
作者:
王思文; 李鹏飞; 王从玉; 鲍思洁; 王勇; 管怀进
226001　江苏省南通市，南通大学附属医院眼科研究所 南通大学医学院
Author(s):
WANG Siwen; LI Pengfei; WANG Congyu; BAO Sijie; WANG Yong; GUAN Huaijin
Department of Ophthalmology,Affiliated Hospital of Nantong University,Medical School of Nantong University,Nantong 226001,Jiangsu Province,China
关键词:
褪黑素; 晶状体上皮细胞; 细胞凋亡; 自噬; 年龄相关性白内障
Keywords:
melatonin;  lens epithelial cells;  apoptosis;  autophagy;  age-related cataract
分类号:
R776.1
DOI:
10.13389/j.cnki.rao.2023.0170
文献标志码:
A
摘要:
目的　探讨褪黑素对H2O2诱导的人晶状体上皮细胞（LECs）氧化损伤的作用机制。
方法　将培养好的人晶状体上皮细胞株（SRA01/04）随机分为对照组（不做任何处理）、H2O2组（400 μmol·L-1 H2O2处理）、DMSO+H2O2组（400 μmol·L-1 H2O2和0.2 μmol·L-1 DSMO处理）、褪黑素+H2O2组（400 μmol·L-1 H2O2和50 μmol·L-1 褪黑素处理）。细胞分组处理后继续培养24 h，进行各项实验室检测比较。CCK8实验检测各组细胞活力；免疫印迹实验检测各组细胞凋亡相关蛋白（Bax和Bcl-2）以及自噬相关蛋白（P62、Beclin-1、LC3）的表达水平；TUNEL法检测各组细胞凋亡情况。
结果　与DMSO+H2O2组相比，褪黑素+H2O2组SRA01/04细胞活力升高，Bax蛋白表达水平降低（P<0.001）， Bcl-2蛋白表达水平增加（P<0.01），SRA01/04细胞凋亡率下降。与DMSO+H2O2组相比，褪黑素+H2O2组SRA01/04细胞中LC3-I、LC3-II及Beclin-1蛋白表达水平均升高（均为P<0.001），P62蛋白表达水平差异无统计学意义（P>0.05），SRA01/04细胞的自噬能力增强。
结论　褪黑素能减轻H2O2诱导的LECs细胞凋亡并增强自噬形成，有望为褪黑素在年龄相关性白内障中的作用机制研究提供新的思路。
Abstract:
Objective　To explore the effect of melatonin on H2O2-induced oxidative damage in human lens epithelial cells (LECs).
Methods　The cultured LECs (SRA01/04) were randomly divided into the control group (without any treatment), H2O2 group (400 μmol·L-1 H2O2 treatment), DMSO+H2O2 group (400 μmol·L-1 H2O2 and 0.2 μmol·L-1 DMSO treatment), and melatonin+H2O2 group (400 μmol·L-1 H2O2 and 50 μmol·L-1 melatonin treatment). After grouping, cells were cultured again for 24 hours. Cell viability was detected by CCK-8. The expression levels of apoptosis-related proteins, such as B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X (Bax), and autophagy-related proteins, such as P62, Beclin-1 and microtubule-associated protein 1A/1B-light chain 3 (LC3), were measured by immunoblotting. Cell apoptosis was detected by TUNEL.
Results　Compared with the DMSO+H2O2 group, the melatonin+H2O2 group showed an increase in SRA01/04 cell viability, a decrease in Bax expression level (P<0.001), an increase in Bcl-2 expression level (P<0.01), and a decrease in SRA01/04 cell apoptosis. Compared with the DMSO+H2O2 group, the expression levels of LC3-I, LC3-II and Beclin-1 proteins in SRA01/04 cells in the melatonin+H2O2 group increased (all P<0.001), while the expression level of P62 showed no significant change (P>0.05), and the autophagy of SRA01/04 cells was enhanced.
Conclusion　Melatonin attenuates H2O2-induced apoptosis and enhances autophagy in LECs, which is expected to provide new ideas for the study of the effect of melatonin on age-related cataract.

参考文献/References:

［1］　SHARMA R,GEORGE A,NIMMAGADDA M,ORTOLAN D,KARLA B S,QURESHY Z,et al.Epithelial phenotype restoring drugs suppress macular degeneration phenotypes in an iPSC model［J］.Nat Commun,2021,12(1):7293.
［2］　PALSAMY P,BIDASEE K R,AYAKI M,AUGUSTEYN R C,CHAN J Y,SHINOHARA T.Methylglyoxal induces endoplasmic reticulum stress and DNA demethylation in the Keap1 promoter of human lens epithelial cells and age-related cataracts［J］.Free Radic Biol Med,2014,72:134-148.
［3］　CROOKE A,HUETE-TORAL F,COLLIGRIS B,PINTOR J.The role and therapeutic potential of melatonin in age-related ocular diseases［J］.J Pineal Res,2017,63(2):e12430.
［4］　CHANG C C,HUANG T Y,CHEN H Y,HUANG T C,LIN L C,CHANG Y J,et al.Protective effect of melatonin against oxidative stress-induced apoptosis and enhanced autophagy in human retinal pigment epithelium cells［J］.Oxid Med Cell Longev,2018,2018:9015765.
［5］　LLEDO V E,ALKOZI H A,SANCHEZ-NAVES J,FERNANDEZ-TORRES M A,GUZMAN-ARANGUEZ A.Modulation of aqueous humor melatonin levels by yellow-filter and its protective effect on lens［J］.J Photochem Photobiol B,2021,221:112248.
［6］　LIU Y C,WILKINS M,KIM T,MALYUGIN B,MEHTA J S.Cataracts［J］.Lancet,2017,390(10094):600-612.
［7］　GALICHANIN K.Exposure to subthreshold dose of UVR-B induces apoptosis in the lens epithelial cells and does not in the lens cortical fibre cells［J］.Acta Ophthalmol,2017,95(8):834-838.
［8］　WU J J,WU W,THOLOZAN F M,SAUNTER C D,GIRKIN J M,QUINLAN R A.A dimensionless ordered pull-through model of the mammalian lens epithelium evidences scaling across species and explains the age-dependent changes in cell density in the human lens［J］.J R Soc Interface,2015,12(108):20150391.
［9］　JI Y,CAI L,ZHENG T,YE H,RONG X,RAO J,et al.The mechanism of UVB irradiation induced-apoptosis in cataract［J］.Mol Cell Biochem,2015,401(1-2):87-95.
［10］　ZHANG L,YAN Q,LIU J P,ZOU L J,LIU J,SUN S,et al.Apoptosis:its functions and control in the ocular lens［J］.Curr Mol Med,2010,10(9):864-875.
［11］　LI W C,KUSZAK J R,DUNN K,WANG R R,MA W,WANG G M,et al.Lens epithelial cell apoptosis appears to be a common cellular basis for non-congenital cataract development in humans and animals［J］.J Cell Biol,1995,130(1):169-181.
［12］　ZHANG F,MENG W,TONG B.Down-regulation of microRNA-133b suppresses apoptosis of lens epithelial cell by up-regulating BCL2L2 in age-related cataracts［J］.Med Sci Monit,2016,22:4139-4145.
［13］　SCHMIDT T M,DO M T,DACEY D,LUCAS R,HATTAR S,MATYNIA A.Melanopsin-positive intrinsically photosensitive retinal ganglion cells:from form to function［J］.J Neurosci,2011,31(45):16094-16101.
［14］　OSTRIN L A.Ocular and systemic melatonin and the influence of light exposure［J］.Clin Exp Optom,2019,102(2):99-108.
［15］　ARGUN M,TOK L,UGUZ A C,CELIK O,TOK O Y,NAZIROGLU M.Melatonin and amfenac modulate calcium entry,apoptosis,and oxidative stress in ARPE-19 cell culture exposed to blue light irradiation (405 nm)［J］.Eye (Lond),2014,28(6):752-760.
［16］　FU Y,TANG M,FAN Y,ZOU H,SUN X,XU X.Anti-apoptotic effects of melatonin in retinal pigment epithelial cells［J］.Front Biosci (Landmark Ed),2012,17(4):1461-1468.
［17］　JIANG T,CHANG Q,ZHAO Z,YAN S,WANG L,CAI J,et al.Melatonin-mediated cytoprotection against hyperglycemic injury in Muller cells［J］.PLoS One,2012,7(12):e50661.
［18］　LLEDO V E,ALKOZI H A,SANCHEZ-NAVES J,FERNANDEZ-TORRES M A,GUZMAN-ARANGUEZ A.Modulation of aqueous humor melatonin levels by yellow-filter and its protective effect on lens［J］.J Photochem Photobiol B,2021,221:112248.
［19］　BAI J,DONG L,SONG Z,GE H,CAI X,WANG G,et al.The role of melatonin as an antioxidant in human lens epithelial cells［J］.Free Radic Res,2013,47(8):635-642.
［20］　ABE M,REITER R J,ORHII P B,HARA M,POEGGELER B.Inhibitory effect of melatonin on cataract formation in newborn rats:evidence for an antioxidative role for melatonin［J］.J Pineal Res,1994,17(2):94-100.
［21］　SINGH R,LETAI A,SAROSIEK K.Regulation of apoptosis in health and disease:the balancing act of BCL-2 family proteins［J］.Nat Rev Mol Cell Biol,2019,20(3):175-193.
［22］　EDLICH F.BCL-2 proteins and apoptosis:recent insights and unknowns［J］.Biochem Biophys Res Commun,2018,500(1):26-34.
［23］　HE H,DONG W,HUANG F.Anti-amyloidogenic and anti-apoptotic role of melatonin in Alzheimer disease［J］.Curr Neuropharmacol,2010,8(3):211-217.
［24］　GONG Y H,HUA N,ZANG X,HUANG T,HE L.Melatonin ameliorates Abeta(1-42) -induced Alzheimer’s cognitive deficits in mouse model［J］.J Pharm Pharmacol,2018,70(1):70-80.
［25］　OZSIMSEK A,OVEY I S.Potential effects of melatonin on TRPA1 channels in the prevention and treatment of alzheimer’s disease［J］.Noro Psikiyatr Ars,2022,59(3):188-192.
［26］　JUNG Y J,CHOI H,OH E.Melatonin attenuates MPP(+)-induced apoptosis via heat shock protein in a Parkinson’s disease model［J］.Biochem Biophys Res Commun,2022,621：59-66.
［27］　KLIONSKY D J,PETRONI G,AMARAVADI R K,BAEHRECKE E H,BALLABIO A,BOYA P,et al.Autophagy in major human diseases［J］.EMBO J,2021,40(19):e108863.
［28］　MAIURI M C,KROEMER G.Therapeutic modulation of autophagy:which disease comes first?［J］.Cell Death Differ,2019,26(4):680-689.
［29］　HUANG J,YU W,HE Q,HE X,YANG M,CHEN W,et al.Autophagy facilitates age-related cell apoptosis-a new insight from senile cataract［J］.Cell Death Dis,2022,13(1):37.
［30］　LAPIERRE L R,KUMSTA C,SANDRI M,BALLABIO A,HANSEN M.Transcriptional and epigenetic regulation of autophagy in aging［J］.Autophagy,2015,11(6):867-880.
［31］　FERNANDEZ A F,SEBTI S,WEI Y,ZOU Z,SHI M,MCMILLAN K L,et al.Disruption of the beclin 1-BCL2 autophagy regulatory complex promotes longevity in mice［J］.Nature,2018,558(7708):136-140.
［32］　GUO Y,WANG J,WANG Z,YANG Y,WANG X,DUAN Q.Melatonin protects N2a against ischemia/reperfusion injury through autophagy enhancement［J］.J Huazhong Univ Sci Technolog Med Sci,2010,30(1):1-7.
［33］　TENG Y C,TAI Y I,HUANG H J,LIN A M.Melatonin ameliorates arsenite-induced neurotoxicity:involvement of autophagy and mitochondria［J］.Mol Neurobiol,2015,52(2):1015-1022.
［34］　TANIDA I,UENO T,KOMINAMI E.LC3 and autophagy［J］.Methods Mol Biol,2008,445:77-88.
［35］　BJORKOY G,LAMARK T,BRECH A,OUTZEN H,PERANDER M,OVERVATN A,et al.p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death［J］.J Cell Biol,2005,171(4):603-614.

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备注/Memo

备注/Memo:
国家自然科学基金面上项目（编号：82171038）
更新日期/Last Update: 2023-11-05