安神定志灵对自发性高血压大鼠前额叶、纹状体多巴胺和去甲肾上腺素含量的影响
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摘要
目的:研究安神定志灵对注意缺陷多动障碍(ADHD)动物模型-自发性高血压大鼠(SHR)前额叶皮质、纹状体脑组织中多巴胺(dopamine,DA)和去甲肾上腺素(norepinephrine,NE)递质含量的影响,探讨该药治疗ADHD的作用机制。方法:40只SHR随机分为5组(模型组、托莫西汀组、安神定志灵低、中、高剂量组),每组8只,Wistar-Kyoto(WKY)大鼠8只作为正常对照组。安神定志灵低、中、高剂量组分别按生药剂量11.85 g/(kg·d),23.7 g/(kg·d),47.4 g/(kg·d)灌胃,每日1次;托莫西汀组按5 mg/(kg·d)给予托莫西汀灌胃,每日1次;模型组和正常对照组每日按体重给予等量生理盐水。实验4周后处死大鼠,采用高效液相色谱-荧光法检测各组大鼠前额叶皮质、纹状体脑组织多巴胺和去甲肾上腺素含量水平。结果:(1)DA递质含量:模型组SHR大鼠前额叶皮质DA含量明显低于正常WKY大鼠(P=0.006),托莫西汀和安神定志灵高剂量均可显著增加DA含量(P<0.05),且两者之间无明显差异(P>0.05);模型组SHR大鼠纹状体部位DA含量明显高于正常WKY大鼠(P=0.000),托莫西汀和安神定志灵各剂量均能显著降低DA含量(P=0.000),且两者之间无明显统计学差异(P>0.05)。(2)NE递质含量:模型组SHR大鼠前额叶皮质NE含量明显低于正常组WKY大鼠(P=0.006),托莫西汀组、安神定志灵中剂量、高剂量组均能显著增加NE含量(P<0.05),且两两比较无统计学差异(P>0.05);模型组SHR大鼠纹状体NE的含量与正常组WKY大鼠无统计学差异(P>0.05)。结论:SHR大鼠前额叶皮质和纹状体存在NE/DA递质失调,前额叶皮质主要表现为NE/DA递质含量的降低,纹状体主要表现为DA含量的增高,与ADHD发病有关;托莫西汀及安神定志灵均能调节NE/DA含量的失衡,且两者之间无明显差异。
Objective: To study the effects of Anshen Dingzhi Ling( ADL) on content of dopamine( DA) and norepinephrine( NE) in prefrontal cortex and striatum of spontaneously hypertensive rat( SHR) and detect the mechanism of ADL treating ADHD. Methods: Twenty- five SHR rats were randomly divided into 5 groups including the model group,atomoxetine group,ADL low dose group,ADL medium dose group and ADL high dose group,meanwhile 5 Wistar- Kyoto( WKY) rats were chosen as the control group. The dosages of three groups of ADL were 11. 85 g/( kg·d),23. 7 g/( kg·d),47. 4 g/( kg·d),ig,and the atomoxetine group were given atomoxetine 5mg/( kg·d) once a day. Rats in the model group and normal control group were perfused with the same volume of normal saline as the criteria of body mass. After 4 weeks,the rats in experiment were killed and separated the prefrontal cortex and striatum brain tissues. Finally,we used Fluorescent- HPLC method to detect the content level of dopamine and norepinephrine in both brain parts. Results:( 1) DA content: The DA content of the model group in the prefrontal cortex was obviously lower than that of the control group( P = 0. 006). Compared with the model group,the DA content was significantly increased in the atomoxetine group and ADL high dosage group( P < 0. 05),and there was no significant statistical difference between the two groups( P > 0. 05). The DA content of the model group in the striatum was obviously higher than that of the control group( P = 0. 000). Compared to the model group,the DA content was significantly decreased in the atomoxetine group and every ADL dosage group( P = 0. 000),and there were no significant statistical differences among these groups( P > 0. 05).( 2) NE content: The NE content of the model group in the prefrontal cortex was obviously lower than that of the control group( P = 0. 006).The NE content in the atomoxetine group,ADL medium and high dosage group was significantly higher than that of the model group( P < 0. 05),and there were no significant statistical differences among the three groups( P > 0. 05). There was no significant statistical difference in the NE content of the striatum between the model group and control group( P > 0. 05). Conclusion:There is NE and DA neurotransmitter dysfunction in the prefrontal cortex and striatum of SHR rats compared with WKY rats. The contents of DA and NE in the prefrontal cortex of SHR rats are both lower than that of the WKY rats and the DA content in the striatum is higher than that of the WKY rats which are all concerned with ADHD. There is no significant statistical difference of the function of ATX and ADL in regulating unbalance of the NE and DA.
Objective: To study the effects of Anshen Dingzhi Ling( ADL) on content of dopamine( DA) and norepinephrine( NE) in prefrontal cortex and striatum of spontaneously hypertensive rat( SHR) and detect the mechanism of ADL treating ADHD. Methods: Twenty- five SHR rats were randomly divided into 5 groups including the model group,atomoxetine group,ADL low dose group,ADL medium dose group and ADL high dose group,meanwhile 5 Wistar- Kyoto( WKY) rats were chosen as the control group. The dosages of three groups of ADL were 11. 85 g/( kg·d),23. 7 g/( kg·d),47. 4 g/( kg·d),ig,and the atomoxetine group were given atomoxetine 5mg/( kg·d) once a day. Rats in the model group and normal control group were perfused with the same volume of normal saline as the criteria of body mass. After 4 weeks,the rats in experiment were killed and separated the prefrontal cortex and striatum brain tissues. Finally,we used Fluorescent- HPLC method to detect the content level of dopamine and norepinephrine in both brain parts. Results:( 1) DA content: The DA content of the model group in the prefrontal cortex was obviously lower than that of the control group( P = 0. 006). Compared with the model group,the DA content was significantly increased in the atomoxetine group and ADL high dosage group( P < 0. 05),and there was no significant statistical difference between the two groups( P > 0. 05). The DA content of the model group in the striatum was obviously higher than that of the control group( P = 0. 000). Compared to the model group,the DA content was significantly decreased in the atomoxetine group and every ADL dosage group( P = 0. 000),and there were no significant statistical differences among these groups( P > 0. 05).( 2) NE content: The NE content of the model group in the prefrontal cortex was obviously lower than that of the control group( P = 0. 006).The NE content in the atomoxetine group,ADL medium and high dosage group was significantly higher than that of the model group( P < 0. 05),and there were no significant statistical differences among the three groups( P > 0. 05). There was no significant statistical difference in the NE content of the striatum between the model group and control group( P > 0. 05). Conclusion:There is NE and DA neurotransmitter dysfunction in the prefrontal cortex and striatum of SHR rats compared with WKY rats. The contents of DA and NE in the prefrontal cortex of SHR rats are both lower than that of the WKY rats and the DA content in the striatum is higher than that of the WKY rats which are all concerned with ADHD. There is no significant statistical difference of the function of ATX and ADL in regulating unbalance of the NE and DA.
引文
[1]Polanczyk G,de Lima MS,Horta BL,et al.The worldwide prevalence of ADHD:a systematic review and metaregression analysis[J].Am JPsychiatry,2007,164(6):942-948.
[2]Makris N,Biederman J,Valera EM,et al.Cortical thinning of the attention and executive function networks in adults with attention-deficit/hyperactivity disorder[J].Cereb Cortex,2007,17(6):1364-1375.
[3]Seidman LJ,Vale ra EM,Makris N,et al.Dorsolateral prefrontal and anterior cingulate cortex volumetric abnormalities in adults with attention-deficit/hyperactivity disorder identified by magnetic resonance imaging[J].Biol Psychiatry,2006,60(10):1071-1080.
[4]Biederman J.Attention-deficit/hyperactivity disorder:a selective overview[J].Biol Psychiatry,2005,57(11):1215-1220.
[5]Arnsten AF.Fundamentals of attention-deficit/hyperactivity disorder:Circuits and pathways[J].J Clin Psychiatry,2006,67(suppl 8):7-12.
[6]韩新民,朱先康.安神定志灵治疗儿童多动症58例临床观察[J].河北中医,2004,26(12):898-899.
[7]刘成全,韩新民,朱先康,等.清心平肝、豁痰开窍法治疗注意缺陷多动障碍30例临床研究[J].新中医,2012,42(12):44-46.
[8]刘成全,韩新民,徐建亚,等.安神定志灵对ADHD模型大鼠纹状体多巴胺转运体表达的影响[J].中国实验方剂学杂志,2011,17(23):89-93.
[9]刘成全,韩新民,徐建亚,等.安神定志灵对ADHD模型鼠前额叶皮质和纹状体多巴胺D1、D2受体表达的影响[J].中国实验方剂学杂志,2011,17(7):136-139.
[10]章新辉,韩新民,徐建亚,等.安神定志灵对自发性高血压大鼠脑中多巴胺受体D3、D4、D5基因表达的影响[J].中国实验方剂学杂志,2013,19(10):186-190.
[11]Sagvolden T,Russell VA,Aase H,et al.Rodent models of attentiondeficit/hyperactivity disorder[J].Biol Psychiatry,2005,57(11):1239-1247.
[12]王店云,单进军,韩新民.正交试验法优选安神定志灵的水提取工艺[J].辽宁中医杂志,2014,41(6):1249-1251.
[13]陈奇.中药药理研究方法学[M].北京:人民卫生出版社,2006:33.
[14]魏元锋,汪益涵,张宁,等.HPLC-荧光检测法测定大鼠血浆及脑组织中5中单胺类神经递质[J].中华中医药学刊,2009,27(9):1942-1943.
[15]鲁燕侠,崔佳,蔺兴遥,等.RP-HPLC-荧光检测法测定小鼠脑组织中5种神经递质的含量[J].解放军药学学报,2003,19(4):262-263,268.
[16]Axelrod J.Regulation of the Neurotransmitter Norepinephrine[M].Cambridge:MA:MIT Press,1974.
[17]During MJ,Bean AJ,Roth RH.Effect of CNS stimulants on the in vivo release of the colocalized transmitters,dopamine and neurotensin,from rat prefrontal cortex[J].Neurosci Lett,1992,140(1):129-133.
[18]Volkow ND,Wang G,Fowler JS,et al.Therapeutic doses of oral methylphenidate significantly increase extracellular dopamine in the human brain[J].J Neurosci,2001,21(2):RC121.
[19]Faraone SV,Biederman J,Spencer T,et al.Efficacy of atomoxetine in adult attention deficit/hyperactivity disorder:a drug-placebo response curve analysis[J].Behav Brain Funct,2005,1:16.
[20]Bymaster FP,Katner JS,Nelson DL,et al.Atomoxetine increases extracellular levels of norepinephrine and dopamine in prefrontal cortex of rat:a potential mechanism for efficacy in attention deficit/hyperactivity disorder[J].Neuropsychopharmacology,2002,27(5):699-711.
[21]马融,古今楠,李新民,等.益智宁神颗粒对自发性高血压模型大鼠脑组织去甲肾上腺素、多巴胺的影响[J].天津中医药,2008,25(1):6-9.
[22]胡颖,林忠东,郑飞霞,等.人参皂苷Rg1对SHR大鼠前额叶、纹状体多巴胺和去甲肾上腺素含量的影响[J].中国中医药科技,2012,19(1):41-42.
[23]胡颖,林忠东,郑飞霞.人参皂苷Rg1对SHR大鼠纹状体GDN-FmRNA基因表达和多巴胺含量的影响[J].中华中医药学刊,2012,30(4):862-864.
[24]Sagvolden T,Johansen EB,Wien G,et al.The spontaneously hypertensive rat model of ADHD:the importance of selecting the appropriate reference strain[J].Neuropharmacology,2009,57(7-8):619-26.
[25]Davids E,Zhang K,Tarazi FI,et al.Animal models of attention-deficit hyperactivity disorder[J].Brain Res Brain Res Rev,2003,42(1):1-21.
[26]Askenasy EP,Taber KH,Yang PB,et al.Methylphenidate(Ritalin):behavioral studies in the rat[J].Int J Neurosci,2007,117(6):757-794.
[27]Ferguson SA,Gough BJ,Cada AM.In vivo basal and amphetamineinduced striatal dopamine and metabolite levels are similar in the spontaneously hypertensive,Wistar-Kyoto and Sprague-Dawley male rats[J].Physiol Behav,2003,80(1):109-114.
[28]Ferguson SA,Paule MG,Cada A,et al.Baseline behavior,but not sensitivity to stimulant drugs,differs among spontaneously hypertensive,Wistar-Kyoto,and Sprague-Dawley rat strain[J].Neurotoxicol Teratol,2007,29(5):547-561.
[29]Heal DJ,Smith SL,Kulkarni RS,et al.New perspectives from micro dialysis studies in freely moving,spontaneously hypertensive rats on the pharmacology of drugs for the treatment of ADHD[J].Pharmaco Biochem Behav,2008,90(2):184-197.
[30]Fan X,Hess EJ.D2-like dopa mine receptors mediate the response to amphetamine in a mouse model of ADHD[J].Neurobiol Dis,2007,26(1):201-211.
[31]Nowak P,Bortel A,Dabrowska J,et al.Amphetamine and mC PPeffects on dopamine and serotonin striatal in vivo microdialysates in an animal model of hyperactivity[J].Neurotox Res,2007,11(2):131-144.
[32]de Villiers AS,Russell VA,Sagvolden T,et al.Alpha 2-adrenoceptor mediated inhibition of[3H]dopamine release from nucleus accumbens slices and monoamine levels in a rat model for attentiondeficit hyperactivity disorder[J].Neurochem Res,1995,20(4):427-433.
[2]Makris N,Biederman J,Valera EM,et al.Cortical thinning of the attention and executive function networks in adults with attention-deficit/hyperactivity disorder[J].Cereb Cortex,2007,17(6):1364-1375.
[3]Seidman LJ,Vale ra EM,Makris N,et al.Dorsolateral prefrontal and anterior cingulate cortex volumetric abnormalities in adults with attention-deficit/hyperactivity disorder identified by magnetic resonance imaging[J].Biol Psychiatry,2006,60(10):1071-1080.
[4]Biederman J.Attention-deficit/hyperactivity disorder:a selective overview[J].Biol Psychiatry,2005,57(11):1215-1220.
[5]Arnsten AF.Fundamentals of attention-deficit/hyperactivity disorder:Circuits and pathways[J].J Clin Psychiatry,2006,67(suppl 8):7-12.
[6]韩新民,朱先康.安神定志灵治疗儿童多动症58例临床观察[J].河北中医,2004,26(12):898-899.
[7]刘成全,韩新民,朱先康,等.清心平肝、豁痰开窍法治疗注意缺陷多动障碍30例临床研究[J].新中医,2012,42(12):44-46.
[8]刘成全,韩新民,徐建亚,等.安神定志灵对ADHD模型大鼠纹状体多巴胺转运体表达的影响[J].中国实验方剂学杂志,2011,17(23):89-93.
[9]刘成全,韩新民,徐建亚,等.安神定志灵对ADHD模型鼠前额叶皮质和纹状体多巴胺D1、D2受体表达的影响[J].中国实验方剂学杂志,2011,17(7):136-139.
[10]章新辉,韩新民,徐建亚,等.安神定志灵对自发性高血压大鼠脑中多巴胺受体D3、D4、D5基因表达的影响[J].中国实验方剂学杂志,2013,19(10):186-190.
[11]Sagvolden T,Russell VA,Aase H,et al.Rodent models of attentiondeficit/hyperactivity disorder[J].Biol Psychiatry,2005,57(11):1239-1247.
[12]王店云,单进军,韩新民.正交试验法优选安神定志灵的水提取工艺[J].辽宁中医杂志,2014,41(6):1249-1251.
[13]陈奇.中药药理研究方法学[M].北京:人民卫生出版社,2006:33.
[14]魏元锋,汪益涵,张宁,等.HPLC-荧光检测法测定大鼠血浆及脑组织中5中单胺类神经递质[J].中华中医药学刊,2009,27(9):1942-1943.
[15]鲁燕侠,崔佳,蔺兴遥,等.RP-HPLC-荧光检测法测定小鼠脑组织中5种神经递质的含量[J].解放军药学学报,2003,19(4):262-263,268.
[16]Axelrod J.Regulation of the Neurotransmitter Norepinephrine[M].Cambridge:MA:MIT Press,1974.
[17]During MJ,Bean AJ,Roth RH.Effect of CNS stimulants on the in vivo release of the colocalized transmitters,dopamine and neurotensin,from rat prefrontal cortex[J].Neurosci Lett,1992,140(1):129-133.
[18]Volkow ND,Wang G,Fowler JS,et al.Therapeutic doses of oral methylphenidate significantly increase extracellular dopamine in the human brain[J].J Neurosci,2001,21(2):RC121.
[19]Faraone SV,Biederman J,Spencer T,et al.Efficacy of atomoxetine in adult attention deficit/hyperactivity disorder:a drug-placebo response curve analysis[J].Behav Brain Funct,2005,1:16.
[20]Bymaster FP,Katner JS,Nelson DL,et al.Atomoxetine increases extracellular levels of norepinephrine and dopamine in prefrontal cortex of rat:a potential mechanism for efficacy in attention deficit/hyperactivity disorder[J].Neuropsychopharmacology,2002,27(5):699-711.
[21]马融,古今楠,李新民,等.益智宁神颗粒对自发性高血压模型大鼠脑组织去甲肾上腺素、多巴胺的影响[J].天津中医药,2008,25(1):6-9.
[22]胡颖,林忠东,郑飞霞,等.人参皂苷Rg1对SHR大鼠前额叶、纹状体多巴胺和去甲肾上腺素含量的影响[J].中国中医药科技,2012,19(1):41-42.
[23]胡颖,林忠东,郑飞霞.人参皂苷Rg1对SHR大鼠纹状体GDN-FmRNA基因表达和多巴胺含量的影响[J].中华中医药学刊,2012,30(4):862-864.
[24]Sagvolden T,Johansen EB,Wien G,et al.The spontaneously hypertensive rat model of ADHD:the importance of selecting the appropriate reference strain[J].Neuropharmacology,2009,57(7-8):619-26.
[25]Davids E,Zhang K,Tarazi FI,et al.Animal models of attention-deficit hyperactivity disorder[J].Brain Res Brain Res Rev,2003,42(1):1-21.
[26]Askenasy EP,Taber KH,Yang PB,et al.Methylphenidate(Ritalin):behavioral studies in the rat[J].Int J Neurosci,2007,117(6):757-794.
[27]Ferguson SA,Gough BJ,Cada AM.In vivo basal and amphetamineinduced striatal dopamine and metabolite levels are similar in the spontaneously hypertensive,Wistar-Kyoto and Sprague-Dawley male rats[J].Physiol Behav,2003,80(1):109-114.
[28]Ferguson SA,Paule MG,Cada A,et al.Baseline behavior,but not sensitivity to stimulant drugs,differs among spontaneously hypertensive,Wistar-Kyoto,and Sprague-Dawley rat strain[J].Neurotoxicol Teratol,2007,29(5):547-561.
[29]Heal DJ,Smith SL,Kulkarni RS,et al.New perspectives from micro dialysis studies in freely moving,spontaneously hypertensive rats on the pharmacology of drugs for the treatment of ADHD[J].Pharmaco Biochem Behav,2008,90(2):184-197.
[30]Fan X,Hess EJ.D2-like dopa mine receptors mediate the response to amphetamine in a mouse model of ADHD[J].Neurobiol Dis,2007,26(1):201-211.
[31]Nowak P,Bortel A,Dabrowska J,et al.Amphetamine and mC PPeffects on dopamine and serotonin striatal in vivo microdialysates in an animal model of hyperactivity[J].Neurotox Res,2007,11(2):131-144.
[32]de Villiers AS,Russell VA,Sagvolden T,et al.Alpha 2-adrenoceptor mediated inhibition of[3H]dopamine release from nucleus accumbens slices and monoamine levels in a rat model for attentiondeficit hyperactivity disorder[J].Neurochem Res,1995,20(4):427-433.