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
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摘要
目的观察c-jun氨基末端激酶(JNK)信号转导通路阻断下电针对慢性应激抑郁模型大鼠海马及血清半胱氨酸天冬氨酸蛋白酶-12(caspase-12)表达的影响,探讨针刺抗抑郁的作用机制。方法 SD大鼠随机分为空白组、模型组、针刺组、氟西汀组、模型+二甲基亚砜(DMSO)组、模型+JNK通路阻断剂SP600125(以下简称SP)组、针刺+SP组、氟西汀+SP组,每组9只。除空白组外,其余组均采用慢性应激结合孤养的方法造模。电针干预选取"内关""三阴交"穴,每次20 min,每天1次,治疗21 d;阳性对照药物给予氟西汀5 mg/kg灌胃,治疗21 d。通过旷场实验对大鼠进行行为学评价,采用酶联免疫吸附(ELISA)法检测大鼠海马和血清中caspase-12蛋白表达。结果实验前各组大鼠行为学检测均无统计学差异(P>0.05)。实验第21天,与空白组相比,模型组的水平穿越格数、竖立次数明显降低(P<0.01);与模型组相比,针刺组能提高模型大鼠的水平穿越格数(P<0.05)。与空白组比较,模型组大鼠海马caspase-12蛋白表达明显上升(P<0.01);与模型组比较,模型+DMSO组、氟西汀组、氟西汀+SP组、模型+SP组、针刺组、针刺+SP组大鼠海马caspase-12蛋白表达均下降(P<0.05)。与空白组比较,模型组大鼠血清caspase-12蛋白表达降低;与模型组比较,模型+DMSO组、模型+SP组、氟西汀组、针刺+SP组大鼠血清caspase-12蛋白表达降低,而氟西汀+SP组、针刺组大鼠caspase-12表达升高,但差异均没有统计学意义(P>0.05)。结论针刺可能是通过抑制JNK信号转导通路活性,从而降低慢性应激模型大鼠海马caspase-12蛋白表达,减少海马神经元凋亡,从而发挥抗抑郁效应。
Objective To observe the effects of electroacupuncture( EA) on the expression of central and peripheral cysteine aspartic acid protease-12 via JNK signal transduction pathway in rats with chronic stress depression and to explore anti-depression mechanism of electro-acupuncture. Methods SD rats were randomly divided into blank group,stress model group,acupuncture group,fluoxetine group,model+ DMSO( dimethyl sulfoxide,soluble organic solvent) group,model + JNK pathway blocker SP600125( hereinafter abbreviated as SP) group,acupuncture + SP group,and fluoxetine + SP group( n = 9 in each group). The combination of chronic stress and solitary raising was used to establish chronic unpredictable mild stress model in seven groups except the control group. The investigated intervention was EA at "Neiguan( PC6) "and "Sanyinjiao( SP6) "points,20 min for each session,once a day,for 21 consecutive days. The positive control group was treated with fluoxetine 5 mg/kg by oral gavage for 21 days.Open field test was used to observe the rats' behaviors. The expression of caspase-12 protein in hippocampus and serum was measured by using ELISA. Results Before the experiment,there was no statistic difference in rat behaviors of all groups( P > 0. 05). At Day 21 of the experiment,the horizontal crossing lattice and frequency of vertical movements markedly decreased compared to the blank group( P <0. 01). Compared with the model group,EA increased the horizontal crossing lattice( P < 0. 05). Compared with the blank group,the expression of caspase-12 protein in the model group increased significantly( P < 0. 01). Compared with the model group,the expression of caspase-12 protein in the model +DMSO group,fluoxetine group,fluoxetine + SP group,acupuncture group and SP group all decreased( P< 0. 05). Compared with the blank group,the expression of caspase-12 protein in the model group decreased. Compared with the model group,the expression of caspase-12 protein in the model + DMSO group,model + SP group,fluoxetine group and acupuncture + SP group( P > 0. 05) decreased,while the expression of caspase-12 in fluoxetine + SP group and acupuncture group was higher than that in the control group,but all the differences were not statistically significant( P > 0. 05). Conclusion Acupuncture treatment may reduce the expression of caspase-12 protein in the hippocampus of chronic stress rats by effectively inhibiting the JNK signal transduction pathway activity and reducing the apoptosis of hippocampal neuron,which contributes to its anti-depression mechanism.
Objective To observe the effects of electroacupuncture( EA) on the expression of central and peripheral cysteine aspartic acid protease-12 via JNK signal transduction pathway in rats with chronic stress depression and to explore anti-depression mechanism of electro-acupuncture. Methods SD rats were randomly divided into blank group,stress model group,acupuncture group,fluoxetine group,model+ DMSO( dimethyl sulfoxide,soluble organic solvent) group,model + JNK pathway blocker SP600125( hereinafter abbreviated as SP) group,acupuncture + SP group,and fluoxetine + SP group( n = 9 in each group). The combination of chronic stress and solitary raising was used to establish chronic unpredictable mild stress model in seven groups except the control group. The investigated intervention was EA at "Neiguan( PC6) "and "Sanyinjiao( SP6) "points,20 min for each session,once a day,for 21 consecutive days. The positive control group was treated with fluoxetine 5 mg/kg by oral gavage for 21 days.Open field test was used to observe the rats' behaviors. The expression of caspase-12 protein in hippocampus and serum was measured by using ELISA. Results Before the experiment,there was no statistic difference in rat behaviors of all groups( P > 0. 05). At Day 21 of the experiment,the horizontal crossing lattice and frequency of vertical movements markedly decreased compared to the blank group( P <0. 01). Compared with the model group,EA increased the horizontal crossing lattice( P < 0. 05). Compared with the blank group,the expression of caspase-12 protein in the model group increased significantly( P < 0. 01). Compared with the model group,the expression of caspase-12 protein in the model +DMSO group,fluoxetine group,fluoxetine + SP group,acupuncture group and SP group all decreased( P< 0. 05). Compared with the blank group,the expression of caspase-12 protein in the model group decreased. Compared with the model group,the expression of caspase-12 protein in the model + DMSO group,model + SP group,fluoxetine group and acupuncture + SP group( P > 0. 05) decreased,while the expression of caspase-12 in fluoxetine + SP group and acupuncture group was higher than that in the control group,but all the differences were not statistically significant( P > 0. 05). Conclusion Acupuncture treatment may reduce the expression of caspase-12 protein in the hippocampus of chronic stress rats by effectively inhibiting the JNK signal transduction pathway activity and reducing the apoptosis of hippocampal neuron,which contributes to its anti-depression mechanism.
引文
[1]Smith K.Mental health:a world of depression[J].Nature,2014,515(7526):181.
[2]吴迁,王思涵,马学红,等.电针抗抑郁临床疗效与中医症状改善的相关性研究[J].针灸临床杂志,2015,9(31):4-7.Wu Q,Wang SH,Ma XH,et al.Correlation between electro-acupuncture for depression and improvement of Chinese medicine symptoms[J].JCAM,2015,9(31):4-7.
[3]王增平,辛莉.针刺合逍遥散治疗抑郁症25例[J].福建中医药,2016,47(3):58.Wang ZP,Xin L.Acupuncture combined with xiaoyao powder in the treatment of depression in 25 cases[J].Fujian Journal of TCM,2016,47(3):58.
[4]Kim I,Xu W,Reed JC.Cell death and endoplasmic reticulum stress:disease relevance and therapeutic opportunities[J].Nat Rev Drug Discov,2008,7(12):1013-1030.
[5]张丽萍,武丽,张曼.解郁1号对抑郁模型大鼠海马神经元细胞凋亡的影响[J].辽宁中医杂志,2009,36(10):1805-1806.Zhang LP,Wu L,Zhang M.Jieyu 1 on apoptosis of hippocampal neurons in depression model rats[J].Liaoning Journal of Traditional Chinese Medicine,2009,36(10):1805-1806.
[6]李文迅,韩焱晶,陶娟,等.电针对抑郁模型大鼠海马神经元凋亡的影响[J].中国中医药信息杂志,2005,12(2):33-35.Li WX,Hang YJ,Tao J,et al.The effect of electro-acupuncture on apoptosis of hippocampal neurons in depression model rats[J].Chinese Journal of Information on TCM,2005,12(2):33-35.
[7]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.Dai W,Li WD,Lu J,et al.Effect of electroacupuncture on hippocampal apoptosis and JNK signal pathway in chronic stress depression rats[J].Acupuncture Research,2010,35(5):330-334.
[8]Willner P.Chronic mild stress(CMS)revisited:consistency and behavioural-neurobiological concordance in the effects of CMS[J].Neuropsychobiology,2005,52(2):90-110.
[9]张旭辉.基于表观遗传学及PKA信号通路探讨针刺对BDNF蛋白及基因表达的影响[D].北京:北京中医药大学,2016.Zhang XH.Effect of acupuncture on BDNF protein and gene expression based on epigenetics and PKA signaling pathway[D].Beijing:Beijing University of Chinese Medicine,2016.
[10]张露芬.实验针灸学[M].北京:化学工业出版社,2010:221.Zhang LF.Experimental acupuncture science[M].Beijing:Chemical Industry Press,2010:221.
[11]于文艳,陈向华.SP600125-JNK抑制剂对缺血再灌注损伤保护作用的研究进展[J].实用医药杂志,2016,4(33):360-362.Yu WY,Chen XH.Research progress on the protective effects of SP600125-JNK inhibitor on ischemia-reperfusion injury[J].Prac J Med&Pharm,2016,4(33):360-362.
[12]沈忠飞,王志坚,潘巍巍,等.氟西汀调控CUMS抑郁大鼠海马突触重塑[J].中国病理生理杂志,2016,32(9):1642-1647.Shen ZF,Wang ZJ,Pan WW,et al.Fluoxetine regulates hippocampal synaptic plasticity in CUMS depression rats[J].Chinese Journal of Pathophysiology,2016,32(9):1642-1647.
[13]Wang YM,Xiao ZJ,Liu XD,et al.Venlafaxine modulates depression-induced behaviour and the expression of Bax mRNA and Bcl-xl mRNA in both hippocampus and myocardium[J].Hum Psychopharmacol,2011,26(2):95-101.
[14]Pahl HL.Signal transduction from the endoplasmic reticulum to the cell nucleus[J].Physiol Rev,1999,79(3):683-701.
[15]Shore GC,Papa FR,Oakes SA.Signaling cell death from the endoplasmic reticulum stress response[J].Curr Opin Cell Biol,2011,23(2):143-149.
[16]Cheng YC,Chang JM,Chen CA,et al.Autophagy modulates endoplasmic reticulum stress-induced cell death in podocytes:a protective role[J].Exp Biol Med(Maywood),2015,240(4):467-476.
[17]Xu LY,Su L,Liu XG.PKCδregulates death receptor 5expression induced by PS-341 through ATF4-ATF3/CHOP axis in human lung cancer cells[J].Mol Cancer Ther,2012,11(10):2174-2182.
[18]Hitomi J,Katayama T,Taniguchi M,et al.Apoptosis induced by endoplasmic reticulum stress depends on activation of caspase-3 via caspase-12[J].Neurosci Lett,2003,357(2):127-130.
[19]Hu P,Han Z,Couvillon AD,et al.Autocrine tumor necrosis factor alpha links endoplasmic reticulum stress to the membrane death receptor pathway through IRE1alpha-mediated NF-kappa B activation and down-regulation of TRAF2 expression[J].Mol Cell Biol,2006,26(8):3071-3084.
[20]Tabas I,Ron D.Integrating the mechanisms of apoptosis induced by endoplasmic reticulum stress[J].Nat Cell Biol,2011,13(3):184-190.
[21]张黎恒.针刺对抑郁模型大鼠海马GLU/GABA兴奋/抑制调节系统的作用研究[D].昆明:云南中医学院,2016.Zhang LH.Study on the effects of excitement and inhibition of depression model rats hippocampal Glu/GABA by acupuncture therapy[D].Kunming:Yunnan University of TCM,2016.
[22]张建斌,王玲玲,吕梅,等.针刺不同腧穴对抑郁症模型大鼠行为学的影响[J].中国针灸,2005,25(9):639-643.Zhang JB,Wang LL,LYU M,et al.Effects of acupuncture at different acupoints on behaviors on depression model rats[J].Chinese Acupuncture&Moxibustion,2005,25(9):639-643.
[2]吴迁,王思涵,马学红,等.电针抗抑郁临床疗效与中医症状改善的相关性研究[J].针灸临床杂志,2015,9(31):4-7.Wu Q,Wang SH,Ma XH,et al.Correlation between electro-acupuncture for depression and improvement of Chinese medicine symptoms[J].JCAM,2015,9(31):4-7.
[3]王增平,辛莉.针刺合逍遥散治疗抑郁症25例[J].福建中医药,2016,47(3):58.Wang ZP,Xin L.Acupuncture combined with xiaoyao powder in the treatment of depression in 25 cases[J].Fujian Journal of TCM,2016,47(3):58.
[4]Kim I,Xu W,Reed JC.Cell death and endoplasmic reticulum stress:disease relevance and therapeutic opportunities[J].Nat Rev Drug Discov,2008,7(12):1013-1030.
[5]张丽萍,武丽,张曼.解郁1号对抑郁模型大鼠海马神经元细胞凋亡的影响[J].辽宁中医杂志,2009,36(10):1805-1806.Zhang LP,Wu L,Zhang M.Jieyu 1 on apoptosis of hippocampal neurons in depression model rats[J].Liaoning Journal of Traditional Chinese Medicine,2009,36(10):1805-1806.
[6]李文迅,韩焱晶,陶娟,等.电针对抑郁模型大鼠海马神经元凋亡的影响[J].中国中医药信息杂志,2005,12(2):33-35.Li WX,Hang YJ,Tao J,et al.The effect of electro-acupuncture on apoptosis of hippocampal neurons in depression model rats[J].Chinese Journal of Information on TCM,2005,12(2):33-35.
[7]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.Dai W,Li WD,Lu J,et al.Effect of electroacupuncture on hippocampal apoptosis and JNK signal pathway in chronic stress depression rats[J].Acupuncture Research,2010,35(5):330-334.
[8]Willner P.Chronic mild stress(CMS)revisited:consistency and behavioural-neurobiological concordance in the effects of CMS[J].Neuropsychobiology,2005,52(2):90-110.
[9]张旭辉.基于表观遗传学及PKA信号通路探讨针刺对BDNF蛋白及基因表达的影响[D].北京:北京中医药大学,2016.Zhang XH.Effect of acupuncture on BDNF protein and gene expression based on epigenetics and PKA signaling pathway[D].Beijing:Beijing University of Chinese Medicine,2016.
[10]张露芬.实验针灸学[M].北京:化学工业出版社,2010:221.Zhang LF.Experimental acupuncture science[M].Beijing:Chemical Industry Press,2010:221.
[11]于文艳,陈向华.SP600125-JNK抑制剂对缺血再灌注损伤保护作用的研究进展[J].实用医药杂志,2016,4(33):360-362.Yu WY,Chen XH.Research progress on the protective effects of SP600125-JNK inhibitor on ischemia-reperfusion injury[J].Prac J Med&Pharm,2016,4(33):360-362.
[12]沈忠飞,王志坚,潘巍巍,等.氟西汀调控CUMS抑郁大鼠海马突触重塑[J].中国病理生理杂志,2016,32(9):1642-1647.Shen ZF,Wang ZJ,Pan WW,et al.Fluoxetine regulates hippocampal synaptic plasticity in CUMS depression rats[J].Chinese Journal of Pathophysiology,2016,32(9):1642-1647.
[13]Wang YM,Xiao ZJ,Liu XD,et al.Venlafaxine modulates depression-induced behaviour and the expression of Bax mRNA and Bcl-xl mRNA in both hippocampus and myocardium[J].Hum Psychopharmacol,2011,26(2):95-101.
[14]Pahl HL.Signal transduction from the endoplasmic reticulum to the cell nucleus[J].Physiol Rev,1999,79(3):683-701.
[15]Shore GC,Papa FR,Oakes SA.Signaling cell death from the endoplasmic reticulum stress response[J].Curr Opin Cell Biol,2011,23(2):143-149.
[16]Cheng YC,Chang JM,Chen CA,et al.Autophagy modulates endoplasmic reticulum stress-induced cell death in podocytes:a protective role[J].Exp Biol Med(Maywood),2015,240(4):467-476.
[17]Xu LY,Su L,Liu XG.PKCδregulates death receptor 5expression induced by PS-341 through ATF4-ATF3/CHOP axis in human lung cancer cells[J].Mol Cancer Ther,2012,11(10):2174-2182.
[18]Hitomi J,Katayama T,Taniguchi M,et al.Apoptosis induced by endoplasmic reticulum stress depends on activation of caspase-3 via caspase-12[J].Neurosci Lett,2003,357(2):127-130.
[19]Hu P,Han Z,Couvillon AD,et al.Autocrine tumor necrosis factor alpha links endoplasmic reticulum stress to the membrane death receptor pathway through IRE1alpha-mediated NF-kappa B activation and down-regulation of TRAF2 expression[J].Mol Cell Biol,2006,26(8):3071-3084.
[20]Tabas I,Ron D.Integrating the mechanisms of apoptosis induced by endoplasmic reticulum stress[J].Nat Cell Biol,2011,13(3):184-190.
[21]张黎恒.针刺对抑郁模型大鼠海马GLU/GABA兴奋/抑制调节系统的作用研究[D].昆明:云南中医学院,2016.Zhang LH.Study on the effects of excitement and inhibition of depression model rats hippocampal Glu/GABA by acupuncture therapy[D].Kunming:Yunnan University of TCM,2016.
[22]张建斌,王玲玲,吕梅,等.针刺不同腧穴对抑郁症模型大鼠行为学的影响[J].中国针灸,2005,25(9):639-643.Zhang JB,Wang LL,LYU M,et al.Effects of acupuncture at different acupoints on behaviors on depression model rats[J].Chinese Acupuncture&Moxibustion,2005,25(9):639-643.