针刺对慢性束缚应激抑郁模型大鼠海马凋亡相关因子的影响
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摘要
目的:观察针刺对抑郁模型大鼠海马细胞色素C、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、凋亡诱导因子(AIF)表达的影响,从凋亡途径探讨针刺防治心理应激引起的早期抑郁的机制。方法:SD大鼠随机分成空白组、模型组、针刺组、氟西汀组,每组8只。应用慢性束缚结合孤养方法建立慢性心理应激抑郁大鼠模型。针刺组于每日应激前1h针刺"百会""印堂"和双侧"三阴交",每日1次,每次20min,持续28d;氟西汀组于每日应激前1h用盐酸氟西汀混悬液(1mL/100g,0.18mg/mL)灌胃,每日1次,持续28d。以体质量和糖水偏好实验、旷场实验评价抑郁模型大鼠行为学改变情况;用Western blot法检测大鼠海马组织细胞色素C、caspase-3和AIF的蛋白表达水平;用DCFH-DA荧光探针技术检测大鼠海马组织活性氧(ROS)的含量。结果:实验结束后,模型组大鼠体质量、糖水偏好指数、旷场实验水平穿越格数和垂直竖立次数均低于空白组(P<0.01)。与模型组比较,针刺组和氟西汀组大鼠体质量、糖水偏好指数上升,水平穿越格数和垂直竖立次数增加(P<0.01,P<0.05)。与空白组比较,模型组大鼠海马组织ROS、细胞色素C、caspase-3和AIF的表达水平显著升高(P<0.01)。与模型组比较,针刺组和氟西汀组大鼠海马组织ROS、细胞色素C、caspase-3和AIF水平显著降低(P<0.01)。结论:针刺干预能显著降低抑郁模型大鼠海马组织凋亡途径的关键因子细胞色素C、caspase-3和AIF的蛋白表达水平,其机制可能与下调线粒体ROS含量有关,这可能是针刺发挥抗抑郁效应的重要途径之一。
Objective To investigate the effect of acupuncture on behavior changes and the expression of reactive oxygen species(ROS),cytochrome C,cysteine-containing aspartate-specific proteases-3(caspase-3),apoptosis inducing factor(AIF)in the hippocampus of rats exposed to chronic restraint stress(CRS),so as to provide experimental evidence for the prevention and treatment of depression.Methods Thirty two male SD rats were equally randomized into control,model,acupuncture and medication(Fluoxetine)groups according to the random number table.The depression model was established by chronic psychological stress stimulation(fasting,water deprivation,chronic restraint combined with solitary raising for 28 days).Manual acupuncture stimulation(2 circles/s)was applied to"Baihui"(GV20),"Yintang"(GV29)and bilateral"Sanyinjiao"(SP6)for2 min,followed by retaining the needles for 20 min.Rats in the medication group were treated by gavage of Fluoxetine(0.18 mg/mL,1 mL/100 g body weight).The treatment was conducted once daily for 28 days.The depression-like behavior was evaluated by using open-field test,sucrose preference test,and body weight.The content of ROS of the right hippocampus tissue was detected by using dichloro-dihydro-fluorescein diacetate(DCFH-DA)assay,and the expression levels of cytochrome C,caspase-3 and AIF proteins of hippocampus were determined by using Western blot.Results After 28 days' stress procedure,the depression rats in the model group showed a significant decrease in the numbers of crossing and rearing of open-field test,sucrose preference index and the body weight compared to the control group(P<0.01).The content of ROS and expression levels of cytochrome C,caspase-3 and AIF in the hippocampus of the model group were significantly increased relevant to the control group(all P<0.01).After the intervention and compared with the model group,the decreased levels of the numbers of crossing and rearing,body weight and sucrose preference index,and the up-regulated levels of content of ROS and expression of cytochrome C,caspase-3 and AIF proteins were all reversed in both acupuncture and medication groups(P<0.01,P<0.05),displaying an anti-depressant effect.The effect of acupuncture was significantly superior to that of medication in up-regulating the numbers of crossing and rearing(P<0.01,P<0.05),and in down-regulating the expression of caspase-3(P<0.01).No significant differences were found between the medication and acupuncture groups in increasing the body weight and sucrose preference index,and in down-regulating the expression of cytochrome C and AIF proteins(P>0.05).Conclusion Acupuncture intervention can significantly reduce ROS content and expression level of cytochrome C,caspase-3 and AIF proteins in the hippocampus possibly by inhibiting the chronic psychological stress—hippocampal oxidative stress—mitochondrial apoptotic pathway in psychological stress-induced depression rats,which may contribute to its effect in relieving depression.
Objective To investigate the effect of acupuncture on behavior changes and the expression of reactive oxygen species(ROS),cytochrome C,cysteine-containing aspartate-specific proteases-3(caspase-3),apoptosis inducing factor(AIF)in the hippocampus of rats exposed to chronic restraint stress(CRS),so as to provide experimental evidence for the prevention and treatment of depression.Methods Thirty two male SD rats were equally randomized into control,model,acupuncture and medication(Fluoxetine)groups according to the random number table.The depression model was established by chronic psychological stress stimulation(fasting,water deprivation,chronic restraint combined with solitary raising for 28 days).Manual acupuncture stimulation(2 circles/s)was applied to"Baihui"(GV20),"Yintang"(GV29)and bilateral"Sanyinjiao"(SP6)for2 min,followed by retaining the needles for 20 min.Rats in the medication group were treated by gavage of Fluoxetine(0.18 mg/mL,1 mL/100 g body weight).The treatment was conducted once daily for 28 days.The depression-like behavior was evaluated by using open-field test,sucrose preference test,and body weight.The content of ROS of the right hippocampus tissue was detected by using dichloro-dihydro-fluorescein diacetate(DCFH-DA)assay,and the expression levels of cytochrome C,caspase-3 and AIF proteins of hippocampus were determined by using Western blot.Results After 28 days' stress procedure,the depression rats in the model group showed a significant decrease in the numbers of crossing and rearing of open-field test,sucrose preference index and the body weight compared to the control group(P<0.01).The content of ROS and expression levels of cytochrome C,caspase-3 and AIF in the hippocampus of the model group were significantly increased relevant to the control group(all P<0.01).After the intervention and compared with the model group,the decreased levels of the numbers of crossing and rearing,body weight and sucrose preference index,and the up-regulated levels of content of ROS and expression of cytochrome C,caspase-3 and AIF proteins were all reversed in both acupuncture and medication groups(P<0.01,P<0.05),displaying an anti-depressant effect.The effect of acupuncture was significantly superior to that of medication in up-regulating the numbers of crossing and rearing(P<0.01,P<0.05),and in down-regulating the expression of caspase-3(P<0.01).No significant differences were found between the medication and acupuncture groups in increasing the body weight and sucrose preference index,and in down-regulating the expression of cytochrome C and AIF proteins(P>0.05).Conclusion Acupuncture intervention can significantly reduce ROS content and expression level of cytochrome C,caspase-3 and AIF proteins in the hippocampus possibly by inhibiting the chronic psychological stress—hippocampal oxidative stress—mitochondrial apoptotic pathway in psychological stress-induced depression rats,which may contribute to its effect in relieving depression.
引文
[1]YANG L F,ZHAO Y H,WANG Y C,et al.The effects of psychological stress on depression[J].Curr Neuropharmacol,2015,13(4):494-504.
[2]LIU W,GE T T,LENG Y S,et al.The role of neural plasticity in depression:from hippocampus to prefrontal cortex[J].Neural Plasticity,2017,2017:1-11.
[3]梁佳,李卫东,吴元坪,等.电针对慢性应激抑郁模型大鼠海马神经元凋亡与再生的影响[J].中华中医药杂志,2012,27(4):947-950.
[4]包伍叶,焦爽,卢峻,等.电针对慢性应激抑郁大鼠学习记忆与海马神经元损伤的影响[J].针刺研究,2014,39(2):136-141.
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[6]TYNAN R J,NAICKER S,HINWOOD M,et al.Chronic stress alters the density and morphology of microglia in a subset of stress-responsive brain regions[J].Brain Behav Immun,2010,24(7):1058-1068.
[7]WANG Y,JIANG H,MENG H,et al.Genome-wide transcriptome analysis of hippocampus in rats indicated that TLR/NLRsignaling pathway was involved in the pathogenisis of depressive disorder induced by chronic restraint stress[J].Brain Res Bull,2017,134:195-204.
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[10]KALISCH R,SCHUBERT M,JACOB W,et al.Anxiety and hippocampus volume in the rat[J].Neuropsychopharmacology,2006,31(5):925-932.
[11]VIKAR V,RAUVALA H,IKONEN E.Cognitive deficit and development of motor impairment in a mouse model of Niemann-Pick type C disease[J].Behav Brain Res,2002,132(1):1-10.
[12]WANG L,KONG Q,LI K,et al.Frequency-dependent changes in amplitude of low-frequency oscillations in depression:a resting-state fMRI study[J].Neurosci Lett,2016,614:105-111.
[13]EISENDRATH S J,GILLUNG E,DELUCCHI K L,et al.A randomized controlled trial of mindfulness-based cognitive therapy for treatment-resistant depression[J].Psychother Psychosom,2016,85(2):99-110.
[14]MAHAR I,BAMBICO F R,MECHAWAR N,et al.Stress,serotonin,and hippocampal neurogenesis in relation to depression and antidepressant effects[J].Neurosci Biobehav Rev,2014,38:173-192.
[15]DE PABLOS R M,HERRERA A J,ESPINOSA-OLIVA AM,et al.Chronic stress enhances microglia activation and exacerbates death of nigral dopaminergic neurons under conditions of inflammation[J].J Neuroinflammation,2014,11(1):34.
[16]YANG C,GUO X,WANG G H,et al.Changes in tau phosphorylation levels in the hippocampus and frontal cortex following chronic stress[J].Braz J Med Biol Res,2014,47(3):237-244.
[17]VSQUEZ C E,RIENER R,REYNOLDS E,et al.NMDAreceptor dysregulation in chronic state:apossible mechanism underlying depression with BDNF down-regulation[J].Neurochem Int,2014,79:88-97.
[18]CHAN Y Y,LO W Y,YANG S N,et al.The benefit of combined acupuncture and antidepressant medication for depression:a systematic review and meta-analysis[J].JAffect Disord,2015,176:106-117.
[19]MUKAINO Y,PARK J,WHITE A,et al.The effectiveness of acupuncture for depression:a systematic review of randomised controlled trials[J].Acupunct Med,2005,23(2):70-76.
[20]史榕荇,吴茜,秦丽娜,等.电针百会、印堂对慢性应激模型大鼠体重及HPA轴影响的研究[J].针灸临床杂志,2007,23(1):50-53.
[21]史榕荇,秦丽娜,吴茜,等.电针对慢性应激模型大鼠行为学及海马谷氨酸含量影响的研究[J].北京中医药大学学报,2007,30(3):177-180.
[22]田朝阳,图娅,姜会梨,等.电针对慢性应激抑郁模型大鼠海马及血清caspase-12表达的影响[J].北京中医药大学学报,2018,41(2):154-159.
[23]戴巍,李卫东,卢峻,等.电针对慢性应激抑郁大鼠海马神经元凋亡及JNK信号转导通路的影响[J].针刺研究,2010,35(5):330-334.
[24]NA Y,BING L,LIU Y,et al.Electro-acupuncture alleviates chronic unpredictable stress-induced depressive-and anxiety-like behavior and hippocampal neuroinflammation in rat model of depression[J].Front Mol Neurosci,2018,11:149.
[25]DUAN D M,YANG X Y,TU Y,et al.Hippocampal gene expression in a rat model of depression after electroacupuncture at the Baihui and Yintang acupoints[J].Neural Regen Res,2014,9(1):76-83.
[26]李想,许明敏,姜会梨,等.手针干预对慢性温和不可预知应激模型大鼠前额叶皮层细胞外调节蛋白激酶1/2和脑源性神经营养因子表达的影响[J].针刺研究,2018,43(11):705-710.
[27]董莎,姜会梨,王瑜,等.针刺对慢性束缚应激抑郁大鼠海马及前额叶皮层胶质纤维酸性蛋白和血清白介素-10表达的影响[J].针刺研究,2018,43(4):209-214.
[28]KROEMER G,GALLUZZI L,BRENNER C.Mitochondrial membrane permeabilization in cell death[J].Physiol Rev,2007,87(1):99-163.
[29]LUANPITPONG S,CHANVORACHOTE P,NIMMAN-NIT U,et al.Mitochondrial superoxide mediates doxorubicin-induced keratinocyte apoptosis through oxidative modification of ERK and Bcl-2ubiquitination[J].Biochem Pharmacol,2012,83(12):1643-1654.
[30]尹立国,崔建忠.线粒体通透性转换孔的研究现状[J].华北煤炭医学院学报,2007,9(4):486-487.
[31]KUMARI A,KAKKAR P.Lupeol prevents acetaminopheninduced in vivo hepatotoxicity by altering the Bax/Bcl-2and oxidative stress-mediated mitochondrial signaling cascade[J].Life Sci,2012,90(15-16):561-570.
[32]ZHANG Y F,HE W,ZHANG C,et al.Role of receptor interacting protein(RIP)1on apoptosis-inducing factor-mediated necroptosis during acetaminophen-evoked acute liver failure in mice[J].Toxicol Lett,2014,225(3):445-453.
[33]MOREIRA A C,BRANCO A F,SAMPAIO S F,et al.Mitochondrial apoptosis-inducing factor is involved in doxorubicin-induced toxicity on H9c2cardiomyoblasts[J].Biochim Biophys Acta,2014,1842(12Pt A):2468-2478.
[2]LIU W,GE T T,LENG Y S,et al.The role of neural plasticity in depression:from hippocampus to prefrontal cortex[J].Neural Plasticity,2017,2017:1-11.
[3]梁佳,李卫东,吴元坪,等.电针对慢性应激抑郁模型大鼠海马神经元凋亡与再生的影响[J].中华中医药杂志,2012,27(4):947-950.
[4]包伍叶,焦爽,卢峻,等.电针对慢性应激抑郁大鼠学习记忆与海马神经元损伤的影响[J].针刺研究,2014,39(2):136-141.
[5]MELIA K R,RYABININ A E,SCHROEDER R,et al.Induction and habituation of immediate early gene expression in rat brain by acute and repeated restraint stress[J].J Neurosci,1994,14(10):5929-5938.
[6]TYNAN R J,NAICKER S,HINWOOD M,et al.Chronic stress alters the density and morphology of microglia in a subset of stress-responsive brain regions[J].Brain Behav Immun,2010,24(7):1058-1068.
[7]WANG Y,JIANG H,MENG H,et al.Genome-wide transcriptome analysis of hippocampus in rats indicated that TLR/NLRsignaling pathway was involved in the pathogenisis of depressive disorder induced by chronic restraint stress[J].Brain Res Bull,2017,134:195-204.
[8]张露芬.实验针灸学[M].北京:化学工业出版社,2010:221.
[9]TREADWAY M T,ZALD D H.Reconsidering anhedonia in depression:lessons from translational neuroscience[J].Neurosci Biobehav Rev,2011,35(3):537-555.
[10]KALISCH R,SCHUBERT M,JACOB W,et al.Anxiety and hippocampus volume in the rat[J].Neuropsychopharmacology,2006,31(5):925-932.
[11]VIKAR V,RAUVALA H,IKONEN E.Cognitive deficit and development of motor impairment in a mouse model of Niemann-Pick type C disease[J].Behav Brain Res,2002,132(1):1-10.
[12]WANG L,KONG Q,LI K,et al.Frequency-dependent changes in amplitude of low-frequency oscillations in depression:a resting-state fMRI study[J].Neurosci Lett,2016,614:105-111.
[13]EISENDRATH S J,GILLUNG E,DELUCCHI K L,et al.A randomized controlled trial of mindfulness-based cognitive therapy for treatment-resistant depression[J].Psychother Psychosom,2016,85(2):99-110.
[14]MAHAR I,BAMBICO F R,MECHAWAR N,et al.Stress,serotonin,and hippocampal neurogenesis in relation to depression and antidepressant effects[J].Neurosci Biobehav Rev,2014,38:173-192.
[15]DE PABLOS R M,HERRERA A J,ESPINOSA-OLIVA AM,et al.Chronic stress enhances microglia activation and exacerbates death of nigral dopaminergic neurons under conditions of inflammation[J].J Neuroinflammation,2014,11(1):34.
[16]YANG C,GUO X,WANG G H,et al.Changes in tau phosphorylation levels in the hippocampus and frontal cortex following chronic stress[J].Braz J Med Biol Res,2014,47(3):237-244.
[17]VSQUEZ C E,RIENER R,REYNOLDS E,et al.NMDAreceptor dysregulation in chronic state:apossible mechanism underlying depression with BDNF down-regulation[J].Neurochem Int,2014,79:88-97.
[18]CHAN Y Y,LO W Y,YANG S N,et al.The benefit of combined acupuncture and antidepressant medication for depression:a systematic review and meta-analysis[J].JAffect Disord,2015,176:106-117.
[19]MUKAINO Y,PARK J,WHITE A,et al.The effectiveness of acupuncture for depression:a systematic review of randomised controlled trials[J].Acupunct Med,2005,23(2):70-76.
[20]史榕荇,吴茜,秦丽娜,等.电针百会、印堂对慢性应激模型大鼠体重及HPA轴影响的研究[J].针灸临床杂志,2007,23(1):50-53.
[21]史榕荇,秦丽娜,吴茜,等.电针对慢性应激模型大鼠行为学及海马谷氨酸含量影响的研究[J].北京中医药大学学报,2007,30(3):177-180.
[22]田朝阳,图娅,姜会梨,等.电针对慢性应激抑郁模型大鼠海马及血清caspase-12表达的影响[J].北京中医药大学学报,2018,41(2):154-159.
[23]戴巍,李卫东,卢峻,等.电针对慢性应激抑郁大鼠海马神经元凋亡及JNK信号转导通路的影响[J].针刺研究,2010,35(5):330-334.
[24]NA Y,BING L,LIU Y,et al.Electro-acupuncture alleviates chronic unpredictable stress-induced depressive-and anxiety-like behavior and hippocampal neuroinflammation in rat model of depression[J].Front Mol Neurosci,2018,11:149.
[25]DUAN D M,YANG X Y,TU Y,et al.Hippocampal gene expression in a rat model of depression after electroacupuncture at the Baihui and Yintang acupoints[J].Neural Regen Res,2014,9(1):76-83.
[26]李想,许明敏,姜会梨,等.手针干预对慢性温和不可预知应激模型大鼠前额叶皮层细胞外调节蛋白激酶1/2和脑源性神经营养因子表达的影响[J].针刺研究,2018,43(11):705-710.
[27]董莎,姜会梨,王瑜,等.针刺对慢性束缚应激抑郁大鼠海马及前额叶皮层胶质纤维酸性蛋白和血清白介素-10表达的影响[J].针刺研究,2018,43(4):209-214.
[28]KROEMER G,GALLUZZI L,BRENNER C.Mitochondrial membrane permeabilization in cell death[J].Physiol Rev,2007,87(1):99-163.
[29]LUANPITPONG S,CHANVORACHOTE P,NIMMAN-NIT U,et al.Mitochondrial superoxide mediates doxorubicin-induced keratinocyte apoptosis through oxidative modification of ERK and Bcl-2ubiquitination[J].Biochem Pharmacol,2012,83(12):1643-1654.
[30]尹立国,崔建忠.线粒体通透性转换孔的研究现状[J].华北煤炭医学院学报,2007,9(4):486-487.
[31]KUMARI A,KAKKAR P.Lupeol prevents acetaminopheninduced in vivo hepatotoxicity by altering the Bax/Bcl-2and oxidative stress-mediated mitochondrial signaling cascade[J].Life Sci,2012,90(15-16):561-570.
[32]ZHANG Y F,HE W,ZHANG C,et al.Role of receptor interacting protein(RIP)1on apoptosis-inducing factor-mediated necroptosis during acetaminophen-evoked acute liver failure in mice[J].Toxicol Lett,2014,225(3):445-453.
[33]MOREIRA A C,BRANCO A F,SAMPAIO S F,et al.Mitochondrial apoptosis-inducing factor is involved in doxorubicin-induced toxicity on H9c2cardiomyoblasts[J].Biochim Biophys Acta,2014,1842(12Pt A):2468-2478.
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