氟西汀通过上调海马内溴结构域蛋白4的表达改善慢性束缚应激所致小鼠的抑郁样行为
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摘要
目的探讨氟西汀(FLX)对慢性束缚应激(CRS)所致小鼠抑郁样行为及海马内溴结构域蛋白4(BRD4)表达的影响。方法 24只雄性昆明小鼠随机分为生理盐水对照(NS)组、抑郁模型(CRS)组、氟西汀干预(CRS+FLX)组。慢性束缚应激3周建立小鼠抑郁模型,应激的第8天至第21天CRS+FLX组于应激前30 min腹腔注射氟西汀(10 mg/kg),NS组及CRS组注射等体积生理盐水。采用糖水偏好实验、喷糖实验、强迫游泳实验、新旧事物识别实验和旷场实验检测各组小鼠行为变化;采用Western blotting及Real-time PCR法检测小鼠海马BRD4蛋白和mRNA的表达情况。结果与NS组相比,CRS组小鼠表现出明显的抑郁样行为,包括糖水偏好百分比显著降低(P<0.01),喷糖实验舔糖时间缩短(P<0.05),强迫游泳不动时间增加(P<0.01),新事物辨别指数降低(P<0.0001),抗抑郁药FLX干预可逆转CRS所诱导的上述抑郁样行为表现(P<0.05);与NS组相比,CRS组小鼠海马BRD4蛋白及mRNA的表达明显下调(1.0000±0.04577比0.08337±0.01658; 1.0000±0.04379比0.6672±0.03193,P<0.05),而FLX可上调抑郁小鼠海马BRD4蛋白及mRNA的表达(0.08337±0.01658比0.4983±0.08574; 0.6672±0.03193比0.8572±0.03181,P<0.05)。结论氟西汀可能通过上调海马BRD4的表达改善小鼠抑郁样行为。
Objective To investigate the effect of fluoxetine(FLX) on the expression of bromodomain-containing protein 4(BRD4) in the hippocampus induced by chronic restraint stress(CRS),with the depression-like behaviour also being determined.Methods Twenty-four male Kunming mice were randomly divided into normal saline(NS) group,CRS group and fluoxetine(FLX) intervention(CRS+FLX) group.The mice of the CRS group were subjected to 3 weeks chronic restraint stress.The mice of CRS + FLX group were treated with fluoxetine by intraperitoneal injection 30 minutes beforerestraint stress from the 8 to 21 days.The mice in the NS group and CRS group were treated with NS.Depression-like behavior was determined by sucrose preference test,sucrose splash test,forced swimming test,novelty object recognition test and open field test.Expression of BRD4 in the hippocampus was determined by Western blotting and Real-time PCR.Results Compared with NS group,mice in the CRS group showed reduced sucrose preference(P<0.01) and decrease the time spent licking in Sucrose splash test(P<0.05),increase immobility time in the forced swimming test(P<0.01) and decrease on the discrimination ratio in the novelty object recognition test,while FLX treatment effectively reversed the depression-like behavior induced by CRS(P < 0.05).Compared with the NS group,CRS led to significantly decreased expression of BRD4 protein and mRNA in the hippocampus(1.000 ± 0.04577 vs 0.08337 ± 0.01658;1.000 ± 0.04379 vs 0.6672 ± 0.03193,P<0.05),which was attenuated by FLX treatment(0.08337 ± 0.01658 vs 0.4983 ± 0.08574;0.6672 ± 0.03193 vs 0.8572 ± 0.03181,P< 0.05).Conclusion Our finding indicates that FLX could alleviate CRSinduced depression-like behavior in mice,which might be attributed to the increased expression of BRD4 in the hippocampus.
Objective To investigate the effect of fluoxetine(FLX) on the expression of bromodomain-containing protein 4(BRD4) in the hippocampus induced by chronic restraint stress(CRS),with the depression-like behaviour also being determined.Methods Twenty-four male Kunming mice were randomly divided into normal saline(NS) group,CRS group and fluoxetine(FLX) intervention(CRS+FLX) group.The mice of the CRS group were subjected to 3 weeks chronic restraint stress.The mice of CRS + FLX group were treated with fluoxetine by intraperitoneal injection 30 minutes beforerestraint stress from the 8 to 21 days.The mice in the NS group and CRS group were treated with NS.Depression-like behavior was determined by sucrose preference test,sucrose splash test,forced swimming test,novelty object recognition test and open field test.Expression of BRD4 in the hippocampus was determined by Western blotting and Real-time PCR.Results Compared with NS group,mice in the CRS group showed reduced sucrose preference(P<0.01) and decrease the time spent licking in Sucrose splash test(P<0.05),increase immobility time in the forced swimming test(P<0.01) and decrease on the discrimination ratio in the novelty object recognition test,while FLX treatment effectively reversed the depression-like behavior induced by CRS(P < 0.05).Compared with the NS group,CRS led to significantly decreased expression of BRD4 protein and mRNA in the hippocampus(1.000 ± 0.04577 vs 0.08337 ± 0.01658;1.000 ± 0.04379 vs 0.6672 ± 0.03193,P<0.05),which was attenuated by FLX treatment(0.08337 ± 0.01658 vs 0.4983 ± 0.08574;0.6672 ± 0.03193 vs 0.8572 ± 0.03181,P< 0.05).Conclusion Our finding indicates that FLX could alleviate CRSinduced depression-like behavior in mice,which might be attributed to the increased expression of BRD4 in the hippocampus.
引文
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[16]Uchida S,Hara K,Kobayashi A,et al.Epigenetic status of Gdnf in the ventral striatum determines susceptibility and adaptation to daily stressful events[J].Neuron,2011,69(2):359-372.
[17]Franklin TB,Russig H,Weiss IC,et al.Epigenetic transmission of the impact of early stress across generations[J].Biol Psychiatry,2010,68(5):408-415.
[18]Chiba S,Numakawa T,Ninomiya M,et al.Chronic restraint stress causes anxiety-and depression-like behaviors,downregulates glucocorticoid receptor expression,and attenuates glutamate release induced by brain-derived neurotrophic factor in the prefrontal cortex[J].Prog Neuropsychopharmacol Biol Psychiatry,2012,39(1):112-119.
[19]Sun H,Kennedy PJ,Nestler EJ.Epigenetics of the depressed brain:role of histone acetylation and methylation[J].Neuropsychopharmacology,2013,38(1):124-137.
[20]KrishnanⅤ,Nestler EJ.The molecular neurobiology of depression[J].Nature,2008,455(7215):894-902.
[21]Dawson MA,Kouzarides T.Cancer epigenetics:from mechanism to therapy[J].Cell,2012,150(1):12-27.
[22]Vigneri R,Vigneri P,Frittitta L.Basal insulin and cardiovascular and other outcomes[J].N Engl J Med,2012,367(18):1761-1762,1763-1764.
[23]Eren KE,Ertugrul A.Psychiatric disorders and epigenetics[J].Turk Psikiyatri Derg,2012,23(2):130-140.
[24]Madhani HD,Francis NJ,Kingston RE,et al.Epigenomics:a roadmap,but to where[J]?Science,2008,322(5898):43-44.
[25]Onishchenko N,Karpova N,Sabri F,et al.Long-lasting depression-like behavior and epigenetic changes of BDNF gene expression induced by perinatal exposure to methylmercury[J].JNeurochem,2008,106(3):1378-1387.
[26]Wu T,Pinto HB,Kamikawa YF,et al.The BET family member BRD4 interacts with OCT4 and regulates pluripotency gene expression[J].Stem Cell Reports,2015,4(3):390-403.
[27]Fernandez P,Scaffidi P,Markert E,et al.Transformation resistance in a premature aging disorder identifies a tumorprotective function of BRD4[J].Cell Rep,2014,9(1):248-260.
[2]Berton O,Nestler EJ.New approaches to antidepressant drug discovery:beyond monoamines[J].Nat Rev Neurosci,2006,7(2):137-151.
[3]Reul JM,Chandramohan Y.Epigenetic mechanisms in stressrelated memory formation[J].Psychoneuroendocrinology,2007,32Suppl 1:S21-S25.
[4]Ren C,Zeng L,Zhou MM.Preparation,biochemical analysis,and structure determination of the bromodomain,an acetyl-Lysine binding domain[J].Methods Enzymol,2016,573:321-343.
[5]Korb E,Herre M,Zucker-ScharffⅠ,et al.BET protein Brd4activates transcription in neurons and BET inhibitor Jq1 blocks memory in mice[J].Nat Neurosci,2015,18(10):1464-1473.
[6]Magistri M,Velmeshev D,Makhmutova M,et al.The BET-Bromodomain inhibitor JQ1 reduces inflammation and Tau phosphorylation at Ser396 in the brain of the 3xTg model of Alzheimer's disease[J].Curr Alzheimer Res,2016,13(9):985-995.
[7]Sartor GC,Powell SK,Brothers SP,et al.Epigenetic readers of lysine acetylation regulate cocaine-induced plasticity[J].JNeurosci,2015,35(45):15062-15072.
[8]Sullivan JM,Badimon A,Schaefer U,et al.Autism-like syndrome is induced by pharmacological suppression of BETproteins in young mice[J].J Exp Med,2015,212(11):1771-1781.
[9]Luo YW,Xu Y,Cao WY,et al.Insulin-like growth factor 2mitigates depressive behavior in a rat model of chronic stress[J].Neuropharmacology,2015,89:318-324.
[10]Li K,Shen S,Ji YT,et al.Melatonin augments the effects of fluoxetine on depression-like behavior and hippocampal BDNF-TrkB signaling[J].Neurosci Bull,2018,34(2):303-311.
[11]Zheng X,Ma S,Kang A,et al.Chemical dampening of Ly6C(hi)monocytes in the periphery produces anti-depressant effects in mice[J].Sci Rep,2016,6:19406.
[12]Surget A,Saxe M,Leman S,et al.Drug-dependent requirement of hippocampal neurogenesis in a model of depression and of antidepressant reversal[J].Biol Psychiatry,2008,64(4):293-301.
[13]Zeng JY,Wang Zh,Niu L,et al.Effect of Panax notoginseng total saponin on lipopolysaccharide induced depression-like behavior and expression of microglia in mice[J].Acta Anatomica Sinica,2018(2):166-171.(in Chinese)曾佳玉,王贞,牛磊,等.三七总皂苷对脂多糖诱导小鼠抑郁样行为及脑内小胶质细胞表达的影响[J].解剖学报,2018(2):166-171.
[14]Xu Y,Cao W,Zhou M,et al.Inactivation of BRD7 results in impaired cognitive behavior and reduced synaptic plasticity of the medial prefrontal cortex[J].Behav Brain Res,2015,286:1-10.
[15]Menard C,Hodes GE,Russo SJ.Pathogenesis of depression:Insights from human and rodent studies[J].Neuroscience,2016,321:138-162.
[16]Uchida S,Hara K,Kobayashi A,et al.Epigenetic status of Gdnf in the ventral striatum determines susceptibility and adaptation to daily stressful events[J].Neuron,2011,69(2):359-372.
[17]Franklin TB,Russig H,Weiss IC,et al.Epigenetic transmission of the impact of early stress across generations[J].Biol Psychiatry,2010,68(5):408-415.
[18]Chiba S,Numakawa T,Ninomiya M,et al.Chronic restraint stress causes anxiety-and depression-like behaviors,downregulates glucocorticoid receptor expression,and attenuates glutamate release induced by brain-derived neurotrophic factor in the prefrontal cortex[J].Prog Neuropsychopharmacol Biol Psychiatry,2012,39(1):112-119.
[19]Sun H,Kennedy PJ,Nestler EJ.Epigenetics of the depressed brain:role of histone acetylation and methylation[J].Neuropsychopharmacology,2013,38(1):124-137.
[20]KrishnanⅤ,Nestler EJ.The molecular neurobiology of depression[J].Nature,2008,455(7215):894-902.
[21]Dawson MA,Kouzarides T.Cancer epigenetics:from mechanism to therapy[J].Cell,2012,150(1):12-27.
[22]Vigneri R,Vigneri P,Frittitta L.Basal insulin and cardiovascular and other outcomes[J].N Engl J Med,2012,367(18):1761-1762,1763-1764.
[23]Eren KE,Ertugrul A.Psychiatric disorders and epigenetics[J].Turk Psikiyatri Derg,2012,23(2):130-140.
[24]Madhani HD,Francis NJ,Kingston RE,et al.Epigenomics:a roadmap,but to where[J]?Science,2008,322(5898):43-44.
[25]Onishchenko N,Karpova N,Sabri F,et al.Long-lasting depression-like behavior and epigenetic changes of BDNF gene expression induced by perinatal exposure to methylmercury[J].JNeurochem,2008,106(3):1378-1387.
[26]Wu T,Pinto HB,Kamikawa YF,et al.The BET family member BRD4 interacts with OCT4 and regulates pluripotency gene expression[J].Stem Cell Reports,2015,4(3):390-403.
[27]Fernandez P,Scaffidi P,Markert E,et al.Transformation resistance in a premature aging disorder identifies a tumorprotective function of BRD4[J].Cell Rep,2014,9(1):248-260.