玉米黄素对衣霉素诱导的SH-SY5Y细胞损伤的保护作用
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摘要
研究玉米黄素对衣霉素诱导损伤的神经母细胞瘤细胞(SH-SY5Y)的保护作用。实验分为空白组、损伤组(模型组)、2、5、10μmol/L玉米黄素保护组。用衣霉素(tunicamycin,TM)建立SH-SY5Y细胞损伤模型,噻唑蓝测定细胞存活率,试剂盒法测定培养液中的乳酸脱氢酶(lactate?dehydrogenase,LDH)水平。结果显示:TM质量浓度在2μg/mL以上时,可显著降低SH-SY5Y细胞活性(P<0.01);与TM损伤组相比,2～10?μmol/L玉米黄素预处理可显著提高细胞存活率(P<0.05),降低LDH的释放率,先用TM处理30 min后再加2、5、10?μmol/L的玉米黄素处理24 h的玉米黄素后处理组结果相同。因此,TM通过诱导内质网应激可引起细胞损伤,玉米黄素能够减轻该损伤作用,并且在一定程度上可逆转TM引起的损伤。
This study was designed to evaluate the protective effect of zeaxanthin on SH-SY5 Y cell injury induced by tunicamycin(TM) in vitro. The experiment was divided into control group, model group and treatment groups with different doses of zeaxanthin. Cell viability was measured by methyl thiazolyl tetrazolium(MTT) assay, and lactate dehydrogenase(LDH) in cells and cell culture were determined by a commercial kit. The results showed that TM at concentrations over2 μg/m L could signi?cantly decrease cell viability(P < 0.01). In contrast to the model group, the pretreatment of zeaxanthin in the concentration range of 2–10 μmol/L could increase cell viability(P < 0.05) and suppress TM-induced LDH release in SH-SY5 Y cells. The same results were obtained with the reverse sequence. TM caused cell injury by inducing endoplasmic reticulum stress. This effect was reduced or even abrogated by zeaxanthin.
This study was designed to evaluate the protective effect of zeaxanthin on SH-SY5 Y cell injury induced by tunicamycin(TM) in vitro. The experiment was divided into control group, model group and treatment groups with different doses of zeaxanthin. Cell viability was measured by methyl thiazolyl tetrazolium(MTT) assay, and lactate dehydrogenase(LDH) in cells and cell culture were determined by a commercial kit. The results showed that TM at concentrations over2 μg/m L could signi?cantly decrease cell viability(P < 0.01). In contrast to the model group, the pretreatment of zeaxanthin in the concentration range of 2–10 μmol/L could increase cell viability(P < 0.05) and suppress TM-induced LDH release in SH-SY5 Y cells. The same results were obtained with the reverse sequence. TM caused cell injury by inducing endoplasmic reticulum stress. This effect was reduced or even abrogated by zeaxanthin.
引文
[1]COLUCCI L,BOSCO M,FASANARO A M,et al.Alzheimer’s disease costs:what we know and what we should take into account[J].Journal of Alzheimers Disease,2014,42(4):1311-1324.DOI:10.3233/JAD-131556.
[2]AN S S,PARK S A,BAGYINSZKY E,et al.A genetic screen of the mutations in the Korean patients with early-onset Alzheimer’s disease[J].Clinical Interventions in Aging,2016,11:1817-1822.DOI:10.2147/CIA.S116724.
[3]TAYLOR J P,HARDY J,FISCHBECK K H.Toxic proteins in neurodegenerative disease[J].Science,2002,296:1991-1995.DOI:10.1126/science.106712.
[4]SM D L M.Brain insulin resistance and deficiency as therapeutic targets in Alzheimer’s disease[J].Current Alzheimer Research,2012,9(1):35-66.DOI:10.2174/156720512799015037.
[5]RAO R V,BREDESEN D E.Misfolded proteins,endoplasmic reticulum stress and neurodegeneration[J].Current Opinion in Cell Biology,2004,16(6):653-662.DOI:10.1016/j.ceb.2004.09.012
[6]GERAKIS Y,HETZ C.Emerging roles of ER stress in the etiology and pathogenesis of Alzheimer’s disease[J].The FEBS Journal,2018,285(6):995-1011.DOI:10.1111/febs.14332.
[7]GENEREUX J C,QU S,ZHOU M,et al.Unfolded protein responseinduced ERdj3 secretion links ER stress to extracellular proteostasis[J].EMBO Journal,2015,34(1):4-19.DOI:10.15252/embj.201488896.
[8]SAILAJA G S,BHOOPATHI P,GORANTLA B,et al.Abstract4992:SPARC overexpression induces ER stress leading to autophagymediated apoptosis in neuroblastoma cells[J].Cancer Research,2012,72(Suppl 8):4992.DOI:10.1158/1538-7445.AM2012-4992.
[9]HETZ C,SAXENA S.ER stress and the unfolded protein response in neurodegeneration[J].Nature Reviews Neurology,2017,13(8):477-491.DOI:10.1038/nrneurol.2017.99.
[10]FOUILLET A,LEVET C,VIRGONE A,et al.ER stress inhibits neuronal death by promoting autophagy[J].Autophagy,2012,8(6):915-926.DOI:10.4161/auto.19716.
[11]MUNSHI S,DAHL R.Cytoprotective small molecule modulators of endoplasmic reticulum stress[J].Bioorganic&Medicinal Chemistry,2016,24(11):2382-2388.DOI:10.1016/j.bmc.2016.03.045.
[12]OHTA K,MIZUNO A,LI S,et al.Endoplasmic reticulum stress enhancesγ-secretase activity[J].Biochemical Biophysical Reseaech Communications,2011,416(3/4):362-366.DOI:10.1016/j.bbrc.2011.11.042.
[13]MARESPERLMAN J A,MILLEN A E,FICEK T L,et al.The body of evidence to support a protective role for lutein and zeaxanthin in delaying chronic disease.overview[J].Journal of Nutrition,2002,132(3):518S-524S.
[14]MA L,DOU H L,HUANG Y M,et al.Improvement of retinal function in early age-related macular degeneration after lutein and zeaxanthin supplementation:a randomized,double-masked,placebocontrolled trial[J].American Journal of Ophthalmology,2012,154(4):625-634.DOI:10.1016/j.ajo.2012.04.014.
[15]LAKEY-BEITIA J,DOENS D,JAGADEESH K D,et al.Antiamyloid aggregation activity of novel carotenoids:implications for Alzheimer’s drug discovery[J].Clinical Interventions in Aging,2017,12(6):815-822.DOI:10.2147/CIA.S134605.
[16]KIM H S,KIM T J,YOO Y M.Melatonin combined with endoplasmic reticulum stress induces cell death via the PI3K/Akt/mTOR pathway in B16F10 melanoma cells[J].PLoS ONE,2014,9(3):e92627.DOI:10.1371/journal.pone.0092627.
[17]MOU Z,YUAN Y H,LOU Y X,et al.Bibenzyl compound 20c protects against endoplasmic reticulum stress in tunicamycin-treated PC12 cells in vitro[J].Acta Pharmacologica Sinica,2016,37(12):1525-1533.DOI:10.1038/aps.2016.75.
[18]KAWADA K,IEKUMO T,SAITO R,et al.Aberrant neuronal differentiation and inhibition of dendrite outgrowth resulting from endoplasmic reticulum stress[J].Journal of Neuroscience Research,2014,92(9):1122-1133.DOI:10.1002/jnr.23389.
[19]CHENG T L,CHEN P S,LI R H,et al.Induction of apurinic endonuclease 1 overexpression by endoplasmic reticulum stress in hepatoma cells[J].International Journal of Molecular Sciences,2014,15(7):12442-12457.DOI:10.3390/ijms150712442.
[20]JIAN L,LU Y,LU S,et al.Chemical chaperone 4-phenylbutyric acid protects H9c2 cardiomyocytes from ischemia/reperfusion injury by attenuating endoplasmic reticulum stress-induced apoptosis[J].Molecular Medicine Reports,2016,13(5):4386-4392.DOI:10.3892/mmr.2016.5063.
[21]YANG H,WANG J,SUN J,et al.A new method to effectively and rapidly generate neurons from SH-SY5Y cells[J].Neuroscience Letters,2016,610:43-47.DOI:10.1016/j.neulet.2015.10.047.
[22]EL HAJ M,COELLO Y,KAPOGIANNIS D,et al.Negative prospective memory in Alzheimer’s disease:?“do?not?perform?that?action”[J].Journal of Alzheimer’s Disease,2018,61(2):663-672.DOI:10.3233/JAD-170807.
[23]HAAPASALO A,PIKKARAINEN M,SOININEN H.Alzheimer’s disease:a report from the 7th Kuopio Alzheimer symposium[J].Neurodegenerative Disease Management,2015,5:379-382.DOI:10.2217/nmt.15.31.
[24]LEOUTSAKOS J M,GROSS A L,JONES R N,et al.‘Alzheimer’s Progression Score’:development of a biomarker summary outcome for AD prevention trials[J].The Journal of Prevention of Alzheimer’s Disease,2016,3(4):229-235.DOI:10.14283/jpad.2016.120.
[25]SUOMINEN M H,PURANEN T M,JYVAKORPI S K,et al.Nutritional guidance improves nutrient intake and quality of life,and may prevent falls in aged persons with Alzheimer disease living with a spouse(NuAD trial)[J].Journal of Nutrition Health&Aging,2015,19(9):901-907.DOI:10.1007/s12603-015-0558-0.
[26]CARRASCO-GALLARDO C,FARIAS G A,FUENTES P,et al.Can nutraceuticals prevent Alzheimer’s disease?potential therapeutic role of a formulation containing shilajit and complex B vitamins[J].Archives of Medical Research,2012,43(8):699-704.DOI:10.1016/j.arcmed.2012.10.010.
[27]SCHEPER W,HOOZEMANS J J.The unfolded protein response in neurodegenerative diseases:a neuropathological perspective[J].Acta Neuropathologica,2015,130(3):315-331.DOI:10.1007/s00401-015-1462-8.
[28]PAPANDREOU M A,KANAKIS C D,POLISSIOU M G,et al.Inhibitory activity on amyloid-beta aggregation and antioxidant properties of Crocus sativus stigmas extract and its crocin constituents[J].Journal of Agriccultural and Food Chemistry,2006,54(23):8762-8768.DOI:10.1021/jf061932a.
[29]ONO K,YOSHIIKE Y,TAKASHIMA A,et al.Vitamin A exhibits potent anti-amyloidogenic and fibril-destabilizing effects in vitro[J].Experimental Neurology,2004,189(2):380-392.DOI:10.1016/j.expneurol.2004.05.035.
[30]ZHOU X Y,WANG S S,DING X,et al.Zeaxanthin improves diabetes-induced cognitive deficit in rats through activiting PI3K/AKTsignaling pathway[J].Brain Research Bulletin,2017,132:190-198.DOI:10.1016/j.brainresbull.2017.06.001.
[31]YING C J,CHEN L,WANG S S,et al.Zeaxanthin ameliorates high glucose-induced mesangial cell apoptosis through inhibiting oxidative stress via activating Akt signalling-pathway[J].Biomedicine&Pharmacotherapy,2017,90:796-805.DOI:10.1016/j.biopha.2017.04.013.
[2]AN S S,PARK S A,BAGYINSZKY E,et al.A genetic screen of the mutations in the Korean patients with early-onset Alzheimer’s disease[J].Clinical Interventions in Aging,2016,11:1817-1822.DOI:10.2147/CIA.S116724.
[3]TAYLOR J P,HARDY J,FISCHBECK K H.Toxic proteins in neurodegenerative disease[J].Science,2002,296:1991-1995.DOI:10.1126/science.106712.
[4]SM D L M.Brain insulin resistance and deficiency as therapeutic targets in Alzheimer’s disease[J].Current Alzheimer Research,2012,9(1):35-66.DOI:10.2174/156720512799015037.
[5]RAO R V,BREDESEN D E.Misfolded proteins,endoplasmic reticulum stress and neurodegeneration[J].Current Opinion in Cell Biology,2004,16(6):653-662.DOI:10.1016/j.ceb.2004.09.012
[6]GERAKIS Y,HETZ C.Emerging roles of ER stress in the etiology and pathogenesis of Alzheimer’s disease[J].The FEBS Journal,2018,285(6):995-1011.DOI:10.1111/febs.14332.
[7]GENEREUX J C,QU S,ZHOU M,et al.Unfolded protein responseinduced ERdj3 secretion links ER stress to extracellular proteostasis[J].EMBO Journal,2015,34(1):4-19.DOI:10.15252/embj.201488896.
[8]SAILAJA G S,BHOOPATHI P,GORANTLA B,et al.Abstract4992:SPARC overexpression induces ER stress leading to autophagymediated apoptosis in neuroblastoma cells[J].Cancer Research,2012,72(Suppl 8):4992.DOI:10.1158/1538-7445.AM2012-4992.
[9]HETZ C,SAXENA S.ER stress and the unfolded protein response in neurodegeneration[J].Nature Reviews Neurology,2017,13(8):477-491.DOI:10.1038/nrneurol.2017.99.
[10]FOUILLET A,LEVET C,VIRGONE A,et al.ER stress inhibits neuronal death by promoting autophagy[J].Autophagy,2012,8(6):915-926.DOI:10.4161/auto.19716.
[11]MUNSHI S,DAHL R.Cytoprotective small molecule modulators of endoplasmic reticulum stress[J].Bioorganic&Medicinal Chemistry,2016,24(11):2382-2388.DOI:10.1016/j.bmc.2016.03.045.
[12]OHTA K,MIZUNO A,LI S,et al.Endoplasmic reticulum stress enhancesγ-secretase activity[J].Biochemical Biophysical Reseaech Communications,2011,416(3/4):362-366.DOI:10.1016/j.bbrc.2011.11.042.
[13]MARESPERLMAN J A,MILLEN A E,FICEK T L,et al.The body of evidence to support a protective role for lutein and zeaxanthin in delaying chronic disease.overview[J].Journal of Nutrition,2002,132(3):518S-524S.
[14]MA L,DOU H L,HUANG Y M,et al.Improvement of retinal function in early age-related macular degeneration after lutein and zeaxanthin supplementation:a randomized,double-masked,placebocontrolled trial[J].American Journal of Ophthalmology,2012,154(4):625-634.DOI:10.1016/j.ajo.2012.04.014.
[15]LAKEY-BEITIA J,DOENS D,JAGADEESH K D,et al.Antiamyloid aggregation activity of novel carotenoids:implications for Alzheimer’s drug discovery[J].Clinical Interventions in Aging,2017,12(6):815-822.DOI:10.2147/CIA.S134605.
[16]KIM H S,KIM T J,YOO Y M.Melatonin combined with endoplasmic reticulum stress induces cell death via the PI3K/Akt/mTOR pathway in B16F10 melanoma cells[J].PLoS ONE,2014,9(3):e92627.DOI:10.1371/journal.pone.0092627.
[17]MOU Z,YUAN Y H,LOU Y X,et al.Bibenzyl compound 20c protects against endoplasmic reticulum stress in tunicamycin-treated PC12 cells in vitro[J].Acta Pharmacologica Sinica,2016,37(12):1525-1533.DOI:10.1038/aps.2016.75.
[18]KAWADA K,IEKUMO T,SAITO R,et al.Aberrant neuronal differentiation and inhibition of dendrite outgrowth resulting from endoplasmic reticulum stress[J].Journal of Neuroscience Research,2014,92(9):1122-1133.DOI:10.1002/jnr.23389.
[19]CHENG T L,CHEN P S,LI R H,et al.Induction of apurinic endonuclease 1 overexpression by endoplasmic reticulum stress in hepatoma cells[J].International Journal of Molecular Sciences,2014,15(7):12442-12457.DOI:10.3390/ijms150712442.
[20]JIAN L,LU Y,LU S,et al.Chemical chaperone 4-phenylbutyric acid protects H9c2 cardiomyocytes from ischemia/reperfusion injury by attenuating endoplasmic reticulum stress-induced apoptosis[J].Molecular Medicine Reports,2016,13(5):4386-4392.DOI:10.3892/mmr.2016.5063.
[21]YANG H,WANG J,SUN J,et al.A new method to effectively and rapidly generate neurons from SH-SY5Y cells[J].Neuroscience Letters,2016,610:43-47.DOI:10.1016/j.neulet.2015.10.047.
[22]EL HAJ M,COELLO Y,KAPOGIANNIS D,et al.Negative prospective memory in Alzheimer’s disease:?“do?not?perform?that?action”[J].Journal of Alzheimer’s Disease,2018,61(2):663-672.DOI:10.3233/JAD-170807.
[23]HAAPASALO A,PIKKARAINEN M,SOININEN H.Alzheimer’s disease:a report from the 7th Kuopio Alzheimer symposium[J].Neurodegenerative Disease Management,2015,5:379-382.DOI:10.2217/nmt.15.31.
[24]LEOUTSAKOS J M,GROSS A L,JONES R N,et al.‘Alzheimer’s Progression Score’:development of a biomarker summary outcome for AD prevention trials[J].The Journal of Prevention of Alzheimer’s Disease,2016,3(4):229-235.DOI:10.14283/jpad.2016.120.
[25]SUOMINEN M H,PURANEN T M,JYVAKORPI S K,et al.Nutritional guidance improves nutrient intake and quality of life,and may prevent falls in aged persons with Alzheimer disease living with a spouse(NuAD trial)[J].Journal of Nutrition Health&Aging,2015,19(9):901-907.DOI:10.1007/s12603-015-0558-0.
[26]CARRASCO-GALLARDO C,FARIAS G A,FUENTES P,et al.Can nutraceuticals prevent Alzheimer’s disease?potential therapeutic role of a formulation containing shilajit and complex B vitamins[J].Archives of Medical Research,2012,43(8):699-704.DOI:10.1016/j.arcmed.2012.10.010.
[27]SCHEPER W,HOOZEMANS J J.The unfolded protein response in neurodegenerative diseases:a neuropathological perspective[J].Acta Neuropathologica,2015,130(3):315-331.DOI:10.1007/s00401-015-1462-8.
[28]PAPANDREOU M A,KANAKIS C D,POLISSIOU M G,et al.Inhibitory activity on amyloid-beta aggregation and antioxidant properties of Crocus sativus stigmas extract and its crocin constituents[J].Journal of Agriccultural and Food Chemistry,2006,54(23):8762-8768.DOI:10.1021/jf061932a.
[29]ONO K,YOSHIIKE Y,TAKASHIMA A,et al.Vitamin A exhibits potent anti-amyloidogenic and fibril-destabilizing effects in vitro[J].Experimental Neurology,2004,189(2):380-392.DOI:10.1016/j.expneurol.2004.05.035.
[30]ZHOU X Y,WANG S S,DING X,et al.Zeaxanthin improves diabetes-induced cognitive deficit in rats through activiting PI3K/AKTsignaling pathway[J].Brain Research Bulletin,2017,132:190-198.DOI:10.1016/j.brainresbull.2017.06.001.
[31]YING C J,CHEN L,WANG S S,et al.Zeaxanthin ameliorates high glucose-induced mesangial cell apoptosis through inhibiting oxidative stress via activating Akt signalling-pathway[J].Biomedicine&Pharmacotherapy,2017,90:796-805.DOI:10.1016/j.biopha.2017.04.013.