吲哚胺2,3-双加氧酶在创伤后应激障碍大鼠海马中的表达变化与作用
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摘要
目的通过检测创伤后应激障碍(post-traumatic stress disorder,PTSD)大鼠海马中吲哚胺2,3-双加氧酶(indoleamine 2,3-dioxygenase,IDO)含量变化与运用IDO抑制剂治疗对神经元细胞的保护机制,探讨PTSD发病原因及机制。方法将60只健康雄性Wistar大鼠随机分为:对照组、PTSD组、PTSD+IDO抑制剂治疗组,运用ELISA试剂盒检测肿瘤坏死因子-α(tumor necrosis factor,TNF-α)、白介素-6(interleukin-6,IL-6)含量变化,通过RT-PCR、Western blot检测IDO的表达情况。通过TUNEL染色法检测海马神经元凋亡率,并对大鼠进行行为学评估。结果检测结果显示对照组、PTSD组与IDO抑制剂治疗组TNF-α分别为[(1.26±0.12)vs.(8.58±0.67)vs.(3.69±0.41)pg/mL]、IL-6分别为[(2.28±0.19)vs.(15.72±1.42)vs.(7.45±0.58)pg/mL]、IDOmRNA分别为[(0.152 7±0.014 7)vs.(0.827 8±0.079 6)vs.(0.223 6±0.038 7)]、IDO蛋白分别为[(0.061 2±0.008 6)vs.(1.232 9±0.114 8)vs.(0.423 5±0.041 1)]、神经元的凋亡率为[(5.46±1.87)%vs.(81.47±6.86)%vs.(42.54±3.98)%](均P<0.05),行为学表现明显改善。结论 PTSD大鼠海马区域TNF-α、IL-6、IDO表达显著提高,IDO抑制剂治疗能降低其含量,细胞因子、IDO在PTSD发病机制中起重要作用,运用IDO抑制剂能改善PTSD大鼠海马区域神经元损伤。
Objective To examine the changes of the expression of indoleamine 2,3-dioxygenase(IDO)in the hippocampus of rats with post-traumatic stress disorder(PTSD)and the protective effect of the IDO inhibitor on neurons,in an attempt to explore the pathogenesis of PTSD.Methods Adult male Wistar rats were randomly divided into three groups:control group,PTSD group,and PTSD+IDO inhibitor group(n=20each).The levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were detected by ELISA and the expression of IDO by RT-PCR and Western blotting.TUNEL staining was used to observe the apoptosis of rat hippocampal neurons.Rat behaviors were also evaluated.Results The levels of TNF-αwere(1.26±0.12),(8.58±0.67),and(3.69±0.41)pg/mL in control,PTSD and PTSD+IDO groups,respectively(P<0.05).The levels of IL-6were(2.28±0.19),(15.72±1.42)and(7.45±0.58)pg/mL in control,PTSD and PTSD+IDO groups,respectively(P<0.05).The expression of IDO was(0.1527±0.0147),(0.8278±0.0796)and(0.2236±0.0387)at mRNA level and(0.0612±0.0086),(1.2329±0.1148),(0.4235±0.0411)at protein level in control,PTSD and PTSD+IDO groups,respectively(P<0.05).The apoptosis rates were(5.46±1.87)%,(81.47±6.86)% and(42.54±3.98)% in control,PTSD and PTSD+IDO groups,respectively(P<0.05).Rat behavioral function was significantly improved in PTSD+IDO group.Conclusion The expression levels of TNF-α,IL-6 and IDO were significantly increased in the hippocampus of PTSD rats,which could be reversed by the IDO inhibitor.The IDO inhibitor could improve the neuron injuries in the hippocampus of PTSD rats.
Objective To examine the changes of the expression of indoleamine 2,3-dioxygenase(IDO)in the hippocampus of rats with post-traumatic stress disorder(PTSD)and the protective effect of the IDO inhibitor on neurons,in an attempt to explore the pathogenesis of PTSD.Methods Adult male Wistar rats were randomly divided into three groups:control group,PTSD group,and PTSD+IDO inhibitor group(n=20each).The levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were detected by ELISA and the expression of IDO by RT-PCR and Western blotting.TUNEL staining was used to observe the apoptosis of rat hippocampal neurons.Rat behaviors were also evaluated.Results The levels of TNF-αwere(1.26±0.12),(8.58±0.67),and(3.69±0.41)pg/mL in control,PTSD and PTSD+IDO groups,respectively(P<0.05).The levels of IL-6were(2.28±0.19),(15.72±1.42)and(7.45±0.58)pg/mL in control,PTSD and PTSD+IDO groups,respectively(P<0.05).The expression of IDO was(0.1527±0.0147),(0.8278±0.0796)and(0.2236±0.0387)at mRNA level and(0.0612±0.0086),(1.2329±0.1148),(0.4235±0.0411)at protein level in control,PTSD and PTSD+IDO groups,respectively(P<0.05).The apoptosis rates were(5.46±1.87)%,(81.47±6.86)% and(42.54±3.98)% in control,PTSD and PTSD+IDO groups,respectively(P<0.05).Rat behavioral function was significantly improved in PTSD+IDO group.Conclusion The expression levels of TNF-α,IL-6 and IDO were significantly increased in the hippocampus of PTSD rats,which could be reversed by the IDO inhibitor.The IDO inhibitor could improve the neuron injuries in the hippocampus of PTSD rats.
引文
[1]殷莉,黄姗姗,何主强.吲哚胺2,3-双加氧酶及细胞因子在创伤后应激障碍患者中的表达变化及意义[J].华中科技大学学报:医学版,2016,45(1):108-110.
[2]Josh M C,Keith B G,Andrew J,et al.Decoding the traumatic memory among women with PTSD:Implications for neurocircuitry models of PTSD and real-time fMRI neurofeedback[J].PLoS One,2015,10(4):1-17.
[3]Pietrzak R H,Goldstein R B,Southwick S M,et al.Psychiatric comorbidity of full and partial post-traumatic stress disorder among older adults in the United States:results from wave 2of the National Epidemiologic Survey on Alcohol and Related Conditions[J].Am J Geriatr Psychiatry,2012,20(5):380-390.
[4]Cohen H,Kaplan Z,Koresh O,et al.Early post-stressor intervention with propranolol is ineffective in preventing post-traumatic stress responses in an animal model for PTSD[J].Eur Neuropsychopharmacol,2011,21(3):230-240.
[5]Thomas C,Eric E,Rachel Y.Biomarkers for combat-related PTSD:focus on molecular networks from high-dimensional data[J].Eur J Psychotraumatol,2014,5(7):1-3.
[6]Moeller D R,Duffy J M,Goolsby A M,et al.Use of a removable mandibular neuroprosthesis for the reduction of post-traumatic stress disorder(PTSD)and mild traumatic brain injury/PTSD/associated nightmares,headaches,and sleep disturbances[J].Spec Oper Med,2014,14(3):64-73.
[7]Koek R J,Langevin J P,Krahl S E,et al.Deep brain stimulation of the basolateral amygdala for treatment-refractory combat post-traumatic stress disorder(PTSD):study protocol for apilot randomized controlled trial with blinded,staggered onset of stimulation[J].Trials,2014,15(5):356-358.
[8]Kousha M,Mehdizadeh T S.Normative life events and PTSD in children:how easy stress can affect children’s brain[J].Acta Med Iran,2013,51(1):47-51.
[9]Richard R.A molecular clue to PTSD[J].PloS Biology,2015,13(2):1-2.
[10]Gao H M,Liu B,Zhang W et al.Novel anti-inflamatory therapy for Parkinson's disease[J].Trends Pharmacol Sci,2003,24(6):395-401.
[11]Hinz B,Brune K.Paracetamol and cyclooxygenase inhibition:is there a cause for concern?[J].Ann Rheum Dis,2012,71(1):20-25.
[12]Said R S,Badr A M,Nada A S,et al.Sodium selenite treatment restores long-lasting ovarian damage induced by irradiation in rats:impact on oxidative stress and apoptosis[J].Reprod Toxicol,2014,43(5):85-93.
[13]Nury T,Zarrouk A,Vejux A,et al.Induction of oxiapoptophagy,a mixed mode of cell death associated with oxidative stress,apoptosis and autophagy,on 7-ketocholesterol-treated158N murine oligodendrocytes:impairment byα-tocopherol[J].Biochem Biophys Res Commun,2014,446(3):714-719.
[14]张建芳,陈欣,王伟,等.异氟烷通过激活Calpain影响大鼠海马神经干细胞的增殖与分化[J].华中科技大学学报:医学版,2015,44(3):263-267.
[15]Puetz T W,Youngstedt S D,Herring M P.Effects of pharmacotherapy on combat-related PTSD,anxiety,and depression:a systematic review and meta-regression analysis[J].PloS One,2015,10(5):529-532.
[2]Josh M C,Keith B G,Andrew J,et al.Decoding the traumatic memory among women with PTSD:Implications for neurocircuitry models of PTSD and real-time fMRI neurofeedback[J].PLoS One,2015,10(4):1-17.
[3]Pietrzak R H,Goldstein R B,Southwick S M,et al.Psychiatric comorbidity of full and partial post-traumatic stress disorder among older adults in the United States:results from wave 2of the National Epidemiologic Survey on Alcohol and Related Conditions[J].Am J Geriatr Psychiatry,2012,20(5):380-390.
[4]Cohen H,Kaplan Z,Koresh O,et al.Early post-stressor intervention with propranolol is ineffective in preventing post-traumatic stress responses in an animal model for PTSD[J].Eur Neuropsychopharmacol,2011,21(3):230-240.
[5]Thomas C,Eric E,Rachel Y.Biomarkers for combat-related PTSD:focus on molecular networks from high-dimensional data[J].Eur J Psychotraumatol,2014,5(7):1-3.
[6]Moeller D R,Duffy J M,Goolsby A M,et al.Use of a removable mandibular neuroprosthesis for the reduction of post-traumatic stress disorder(PTSD)and mild traumatic brain injury/PTSD/associated nightmares,headaches,and sleep disturbances[J].Spec Oper Med,2014,14(3):64-73.
[7]Koek R J,Langevin J P,Krahl S E,et al.Deep brain stimulation of the basolateral amygdala for treatment-refractory combat post-traumatic stress disorder(PTSD):study protocol for apilot randomized controlled trial with blinded,staggered onset of stimulation[J].Trials,2014,15(5):356-358.
[8]Kousha M,Mehdizadeh T S.Normative life events and PTSD in children:how easy stress can affect children’s brain[J].Acta Med Iran,2013,51(1):47-51.
[9]Richard R.A molecular clue to PTSD[J].PloS Biology,2015,13(2):1-2.
[10]Gao H M,Liu B,Zhang W et al.Novel anti-inflamatory therapy for Parkinson's disease[J].Trends Pharmacol Sci,2003,24(6):395-401.
[11]Hinz B,Brune K.Paracetamol and cyclooxygenase inhibition:is there a cause for concern?[J].Ann Rheum Dis,2012,71(1):20-25.
[12]Said R S,Badr A M,Nada A S,et al.Sodium selenite treatment restores long-lasting ovarian damage induced by irradiation in rats:impact on oxidative stress and apoptosis[J].Reprod Toxicol,2014,43(5):85-93.
[13]Nury T,Zarrouk A,Vejux A,et al.Induction of oxiapoptophagy,a mixed mode of cell death associated with oxidative stress,apoptosis and autophagy,on 7-ketocholesterol-treated158N murine oligodendrocytes:impairment byα-tocopherol[J].Biochem Biophys Res Commun,2014,446(3):714-719.
[14]张建芳,陈欣,王伟,等.异氟烷通过激活Calpain影响大鼠海马神经干细胞的增殖与分化[J].华中科技大学学报:医学版,2015,44(3):263-267.
[15]Puetz T W,Youngstedt S D,Herring M P.Effects of pharmacotherapy on combat-related PTSD,anxiety,and depression:a systematic review and meta-regression analysis[J].PloS One,2015,10(5):529-532.