升清降浊制动颗粒对多发性抽动症模型大鼠纹状体内DA水平及DAT、DRD2 mRNA表达的影响
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摘要
目的:观察升清降浊制动颗粒对多发性抽动症(tourette syndrome,TS)模型大鼠纹状体内多巴胺(dopamine,DA)水平及多巴胺转运蛋白(dopamine transporter,DAT)、多巴胺D2受体(dopamine DZ receptor,DRD2) mRNA表达的影响。方法:将70只雄性SD大鼠随机分为空白对照组,模型组,氟哌啶醇组,升清降浊制动颗粒高剂量组、低剂量组,每组14只。采用3,3'-亚氨基二丙腈(3,3'-Iminodipropionitrile,IDPN)制备TS模型大鼠。从造模完成次日开始,空白对照组、模型组给予等体积生理盐水灌胃,氟哌啶醇组、升清降浊制动颗粒高剂量组、升清降浊制动颗粒低剂量组分别给予氟哌啶醇稀释液[200μg·kg-1]、升清降浊制动颗粒高剂量悬液[2. 58 mg·g-1]、升清降浊制动颗粒低剂量悬液[1. 29 mg·g-1]治疗,第21天结束后取材。采用高效液相色谱-四级杆离子阱串联质谱仪(HPLC-MS/MS Q-TRAP)检测纹状体中DA水平,RT-PCR检测纹状体中DAT、DRD2 mRNA表达水平。结果:与空白对照组比较,模型组运动行为评分、刻板行为评分均显著升高,差异均有统计学意义(P <0. 05)。与模型组比较,治疗后高剂量、低剂量升清降浊制动颗粒均能降低IDPN诱导的TS模型大鼠运动行为评分、刻板行为评分,差异均有统计学意义(P <0. 05)。与空白对照组比较,模型组DA水平显著降低,差异有统计学意义(P <0. 05)。与模型组比较,治疗后高剂量、低剂量升清降浊制动颗粒均能升高IDPN诱导的TS模型大鼠DA水平,差异均有统计学意义(P <0. 05)。与空白对照组比较,模型组DAT mRNA表达显著降低,DRD2mRNA表达显著升高,差异均有统计学意义(P <0. 05)。与模型组比较,治疗后高剂量、低剂量升清降浊制动颗粒均能升高DAT mRNA表达,降低DRD2 mRNA表达,差异均有统计学意义(P <0. 05);具有升高纹状体DA含量及上调DAT mRNA、下调DRD2 mRNA表达量的作用(P <0. 05)。结论:升清降浊制动颗粒能改善IDPN诱导的TS模型大鼠行为异常,其机制可能与升高纹状体DA水平及上调DAT mRNA表达、下调DRD2 mRNA表达有关。
Objective: To observe the effect of Shengqing Jiangzhuo Zhidong Granule on dopamine( DA) level,dopamine transporter( DAT) and dopamine D2 in striatum of rats with multiple Tourette syndrome( TS) and on the dopamine DZ receptor( DRD2)mRNA expression. Methods: Seventy male Sprague-Dawley rats were randomly divided into control group,model group,haloperidol group,high-dose group and low-dose group of Shengqing Jiangzhuo Zhidong Granules,with 14 rats in each group. TS model rats were prepared by using 3,3'-iminodipropionitrile( IDPN). From the next day after the modeling was completed,the blank control group and the model group were given an equal volume of normal saline. The haloperidol group,the high-dose group of Shengqing Jiangzhu brake granules,and the low-dose group of Shengqing turbidity brake granules were given separately: Haloperidol dilution[200 μg·kg-1],Shengqing Jiangzhuo Zhidong Granules high-dose suspension [2. 58 mg·g-1],Shengqing Jiangzhuo Zhidong Gradunes low-dose suspension [1. 29 mg·g-1]. Treatment lasted till the end of the 21 st day. The levels of DA in the striatum were detected by high performance liquid chromatography-quadrupole ion trap tandem mass spectrometry( HPLC-MS/MS QTRAP). The expression levels of DAT and DRD2 in the striatum were detected by RT-PCR. Results: Compared with that of the blank control group,the motor behavior scores and stereotype behavior scores of the model group were significantly increased( P <0. 05). Compared with that of the model group,the high-dose and low-dose Shengqing Jiangzhu Zhidong Granules group after treatment could reduce the motor behavior score and stereotype behavior score of the SD model induced by IDPN,and the difference was statistically significant( P < 0. 05). Compared with that of the blank control group,the DA level in the model group was significantly lower( P < 0. 05). Compared with that of the model group,the high-dose and low-dose Shengqing Jiangzhuo Zhidong Granules after treatment could increase the DA level of IDPN-induced TS model rats,and the difference was statistically significant( P < 0. 05). Compared with that of the blank control group,the expression of DAT was significantly decreased and the expression of DRD2 mRNA was significantly increased in the model group( P < 0. 05). Compared with that of the model group,high-dose and low-dose Shengqing Jiangzhuo Zhidong Grenules could increase DAT expression and decrease DRD2 mRNA expression,and the difference was statistically significant( P < 0. 05). Shengqing Jiangzhuo Zhidong Granules had the effect of increasing the content of DA in the striatum and up-regulating DAT and down-regulating the expression of DRD2 mRNA( P < 0. 05). Conclusion:Shengqing Jiangzhuo Zhidong Granules can improve the behavioral abnormality of IDPN-induced TS model rats,which may be related to the increase of striatum DA level,up-regulation of DAT expression and down-regulation of DRD2 mRNA expression.
Objective: To observe the effect of Shengqing Jiangzhuo Zhidong Granule on dopamine( DA) level,dopamine transporter( DAT) and dopamine D2 in striatum of rats with multiple Tourette syndrome( TS) and on the dopamine DZ receptor( DRD2)mRNA expression. Methods: Seventy male Sprague-Dawley rats were randomly divided into control group,model group,haloperidol group,high-dose group and low-dose group of Shengqing Jiangzhuo Zhidong Granules,with 14 rats in each group. TS model rats were prepared by using 3,3'-iminodipropionitrile( IDPN). From the next day after the modeling was completed,the blank control group and the model group were given an equal volume of normal saline. The haloperidol group,the high-dose group of Shengqing Jiangzhu brake granules,and the low-dose group of Shengqing turbidity brake granules were given separately: Haloperidol dilution[200 μg·kg-1],Shengqing Jiangzhuo Zhidong Granules high-dose suspension [2. 58 mg·g-1],Shengqing Jiangzhuo Zhidong Gradunes low-dose suspension [1. 29 mg·g-1]. Treatment lasted till the end of the 21 st day. The levels of DA in the striatum were detected by high performance liquid chromatography-quadrupole ion trap tandem mass spectrometry( HPLC-MS/MS QTRAP). The expression levels of DAT and DRD2 in the striatum were detected by RT-PCR. Results: Compared with that of the blank control group,the motor behavior scores and stereotype behavior scores of the model group were significantly increased( P <0. 05). Compared with that of the model group,the high-dose and low-dose Shengqing Jiangzhu Zhidong Granules group after treatment could reduce the motor behavior score and stereotype behavior score of the SD model induced by IDPN,and the difference was statistically significant( P < 0. 05). Compared with that of the blank control group,the DA level in the model group was significantly lower( P < 0. 05). Compared with that of the model group,the high-dose and low-dose Shengqing Jiangzhuo Zhidong Granules after treatment could increase the DA level of IDPN-induced TS model rats,and the difference was statistically significant( P < 0. 05). Compared with that of the blank control group,the expression of DAT was significantly decreased and the expression of DRD2 mRNA was significantly increased in the model group( P < 0. 05). Compared with that of the model group,high-dose and low-dose Shengqing Jiangzhuo Zhidong Grenules could increase DAT expression and decrease DRD2 mRNA expression,and the difference was statistically significant( P < 0. 05). Shengqing Jiangzhuo Zhidong Granules had the effect of increasing the content of DA in the striatum and up-regulating DAT and down-regulating the expression of DRD2 mRNA( P < 0. 05). Conclusion:Shengqing Jiangzhuo Zhidong Granules can improve the behavioral abnormality of IDPN-induced TS model rats,which may be related to the increase of striatum DA level,up-regulation of DAT expression and down-regulation of DRD2 mRNA expression.
引文
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[9]朱云程,江茜茜,刘丽,等.抽动秽语综合征共患注意缺陷多动障碍男性儿童的反应抑制和精细调节[J].中国心理卫生杂志,2015,29(5):343-348.
[10]郑宏,高国财,郑攀,等.升清降浊制动颗粒治疗多发性抽动症痰热动风证临床研究[J].中医学报,2015,30(11):1650-1652.
[11]郑宏,钱红涛,卢婷婷,等.基于中医传承辅助系统的中医药治疗儿童抽动障碍用药规律研究[J].中国实验方剂学杂志,2016,22(17):233-237.
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[14]张璠.六味地黄丸对多发性抽动症模型大鼠行为及脑多巴胺含量影响的实验研究[D].郑州:河南中医学院,2012.
[15]崔霞,卫利,吴力群,等.健脾止动汤对多发性抽动症模型鼠自主活动的影响及其作用机制的研究[J].北京中医药大学学报(中医临床版),2011,18(1):1-4.
[16]LECKMAN J F,BLOCH M H,SMITH M E,et al. Neurobiological Substrates of Tourette's Disorder[J]. J Child Adolesc Psychop,201020(4):237-247.
[17]VAN SCHOUWENBURG M R,ZWIERS M P,VAN DER SCHAAF M E,ET A L. Anatomical connection strength predicts dopaminergic drug effects on fronto-striatal function[J]. Psychopharmacology(Berl),2013,227(3):521-531.
[18]高汉媛,史正刚,李小芹,等. IDPN诱导TS模型大鼠纹状体DA系统的变化及菖蒲郁金汤的干预作用[J].时珍国医国药,2016,27(6):1367-1369.
[19]谢卉,王珍,季燕,等.红景天苷对Tourette综合征模型大鼠抽动行为的影响[J].中国中西医结合杂志,2016,36(1):90-93.
[20]HERZBERG I,VALENCIADUARTE A V,KAY V A,et al. Association of DRD2 variants and Gilles de la Tourette syndrome in a family-based sample from a South American population isolate[J]. Psychiatric Genetics,2010,20(4):179-183.
[21]JIJUN L,ZAIWANG L,ANYUAN L,et al. Abnormal expression of dopamine and serotonin transporters associated with the pathophysiologic mechanism of Tourette syndrome[J]. Neurology India,2010,58(4):523-529.
[22]郑宏,郑攀,郑启仲.郑启仲运用升降散治疗儿科疾病经验[J].中华中医药杂志,2014,29(6):1864-1866.
[23]郑宏,郑攀,郑启仲.郑启仲治疗小儿多发性抽动症经验[J].中医杂志,2012,63(3):195-197.
[24]钱红涛,郑宏.郑启仲运用升降散规律研究[J].中医学报,2016,31(5):674-678.
[2]KIM D D,WARBURTON D E R,WU N,et al. Effects of physical activity on the symptoms of Tourette syndrome:A systematic review[J].Eur Psych,2017,48(1):13.
[3]MUNASIPOVA S E,ZALYALOVA Z A. Pediatric acute-onset neuropsychiatric syndrome:clinical/laboratory characteristics[J]. Zh Nevrol Psikhiatr Im S S Korsakova,2017,117(11):47-53.
[4]GREENBERG E,TUNG E S,GAUVIN C,et al. Prevalence and predictors of hair pulling disorder and excoriation disorder in Tourette syndrome[J]. Eur Child Adolesc Psychiatry,2018,27(5):569-579.
[5]CUBO E,GONZALEZ C,AUSIN V,et al. The Association of Poor Academic Performance with Tic Disorders:A Longitudinal,Mainstream School-Based Population Study[J]. Neuroepidemiology,2017,48(3/4):155-163.
[6]GIRALDO B O,DAVID M,SANCHEZ Y,et al. Prevalence of Tics Among 6 to 12-Year-Old School children in the Itagui Municipality,Colombia,in 2010[J]. J Child Neurol,2013,28(11):1406-1411.
[7]王道涵.健脾止动汤对TS小鼠纹状体多巴胺系统DR、DAT及谷氨酸系统GLT1的影响[D].北京:北京中医药大学,2013.
[8]侯广舜.宁动颗粒对抽动-秽语综合征模型大鼠纹状体5-羟色胺系统的影响[D].济南:山东大学,2014.
[9]朱云程,江茜茜,刘丽,等.抽动秽语综合征共患注意缺陷多动障碍男性儿童的反应抑制和精细调节[J].中国心理卫生杂志,2015,29(5):343-348.
[10]郑宏,高国财,郑攀,等.升清降浊制动颗粒治疗多发性抽动症痰热动风证临床研究[J].中医学报,2015,30(11):1650-1652.
[11]郑宏,钱红涛,卢婷婷,等.基于中医传承辅助系统的中医药治疗儿童抽动障碍用药规律研究[J].中国实验方剂学杂志,2016,22(17):233-237.
[12]鱼达,李辉,姜骊.实验动物随机分组的计算机程序处理[J].实验动物科学,2011,28(1):25-27.
[13]杜淑娟,彭贝如,程树军,等.抽动-秽语综合征动物模型的建立和评价[J].中国比较医学杂志,2010,20(3):23-25.
[14]张璠.六味地黄丸对多发性抽动症模型大鼠行为及脑多巴胺含量影响的实验研究[D].郑州:河南中医学院,2012.
[15]崔霞,卫利,吴力群,等.健脾止动汤对多发性抽动症模型鼠自主活动的影响及其作用机制的研究[J].北京中医药大学学报(中医临床版),2011,18(1):1-4.
[16]LECKMAN J F,BLOCH M H,SMITH M E,et al. Neurobiological Substrates of Tourette's Disorder[J]. J Child Adolesc Psychop,201020(4):237-247.
[17]VAN SCHOUWENBURG M R,ZWIERS M P,VAN DER SCHAAF M E,ET A L. Anatomical connection strength predicts dopaminergic drug effects on fronto-striatal function[J]. Psychopharmacology(Berl),2013,227(3):521-531.
[18]高汉媛,史正刚,李小芹,等. IDPN诱导TS模型大鼠纹状体DA系统的变化及菖蒲郁金汤的干预作用[J].时珍国医国药,2016,27(6):1367-1369.
[19]谢卉,王珍,季燕,等.红景天苷对Tourette综合征模型大鼠抽动行为的影响[J].中国中西医结合杂志,2016,36(1):90-93.
[20]HERZBERG I,VALENCIADUARTE A V,KAY V A,et al. Association of DRD2 variants and Gilles de la Tourette syndrome in a family-based sample from a South American population isolate[J]. Psychiatric Genetics,2010,20(4):179-183.
[21]JIJUN L,ZAIWANG L,ANYUAN L,et al. Abnormal expression of dopamine and serotonin transporters associated with the pathophysiologic mechanism of Tourette syndrome[J]. Neurology India,2010,58(4):523-529.
[22]郑宏,郑攀,郑启仲.郑启仲运用升降散治疗儿科疾病经验[J].中华中医药杂志,2014,29(6):1864-1866.
[23]郑宏,郑攀,郑启仲.郑启仲治疗小儿多发性抽动症经验[J].中医杂志,2012,63(3):195-197.
[24]钱红涛,郑宏.郑启仲运用升降散规律研究[J].中医学报,2016,31(5):674-678.