摘要
目的构建亚氨基二丙腈(iminodiProPion-itri,IDPN)、2,5-二甲氧-4-碘苯-2-氨基丙烷(5-dimethoxy-4-uodoPhenyl-2-aminoProPan,DOI)、阿扑吗啡(aPomorPhine,APO)3种抽动障碍(tic disorder,TD)大鼠模型对其行为特征、激惹状态及相关递质变化进行比较;选择表观效度及结构效度良好的TD模型分别加载不同应激刺激,构建、评价及筛选最优TD肝阳上亢证病证结合大鼠模型。方法120只SD大鼠随机分为6组:对照组正常饲喂,不做特殊处理;IDPN组腹腔注射IDPN溶液持续1周;DOI组、APO组分别腹腔注射DOI溶液、APO溶液持续2周;APO复合束缚悬吊应激(restraint and suspension stress,RSS)模型组(APO+RSS组)腹腔注射APO溶液2周同时复合慢性束缚悬吊应激持续3周;APO复合声电刺激应激(acoustic and electrical stimulation stress,AESS)模型组(APO+AESS组)腹腔注射APO溶液2周同时复合声电刺激应激持续3周。通过行为学实验评估大鼠刻板运动、自发活动及情绪障碍;酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)法测定纹状体中多巴胺(dopamine,DA)、γ-氨基丁酸(γ-aminobutyric acid,GABA)、谷氨酸(glutamate,GLU)、五羟色胺(serotonin,5-HT)水平变化,血清中血管紧张素II(Angiotensin II,AngⅡ)、精氨酸加压素(arginine vasopressin,AVP)水平变化。结果APO、DOI、IDPN诱导造模结果显示,与对照组比较,3种造模方式均可成功诱导大鼠的自发活动、刻板运动、认知及情绪障碍。IDPN组表现为更明显的刻板运动、认知障碍及焦虑抑郁倾向,与对照组相比DA、GLU水平降低(P<0.05),5-HT水平升高(P<0.05);APO组表现为更明显的自发活动及兴奋、易激惹状态、探索及攻击行为增多,与对照组相比DA、GLU水平升高(P<0.05),AngⅡ水平显著升高(P<0.01);DOI组大鼠行为学表现与APO组相似,但自发活动、刻板运动程度较APO组差;与对照组相比DOI组5-HT水平显著降低(P<0.01)。APO加载复合模型比较结果显示,APO+RSS组大鼠自发活动、刻板运动、探索行为较APO组增加;APO+AESS组表现为更明显的认知障碍。APO+RSS组大鼠纹状体DA、GLU水平、血清AngⅡ较APO组、APO+AESS组增加(P<0.05)。结论APO诱导TD模型表观效度及结构效度良好,并在一定程度上具备中医肝阳上亢证特点;APO复合束缚悬吊应激刺激模型激活RAS系统加重肝阳上亢症状并有利于维持TD模型稳定,是较为理想的TD肝阳上亢证病证结合动物模型。
Objective Three rat TD models,IDPN,DOI and APO,were constructed to compare their behavior characteristics of simulated TD,and changes in relevant transmitters;TD models with good apparent andcharacteristics of hyperactivity of liver yang were selected to load different stress stimuli separately to build composite models,to evaluate and screen the optimal TD hyperactivity of liver yang rat model with integration of disease and syndrome.Methods 120 male Sprague-Dawley rats were randomly divided into six groups:rats in the control group were fed without special treatment;the IDPN group was intraperitoneally injected with IDPN solution for a week;the DOI group and the APO group were intraperitoneally injected with DOI solution and APO solution respectively for 2 weeks;the APO+RSS group was intraperitoneally injected with APO solution for 2 weeks and combined restraint and suspension stress for 3 weeks;the APO+AESS group was intraperitoneally injected with APO solution for 2 weeks and combined acoustic electrical stimulation stress for 3 weeks.Behavioral tests were used to evaluate the stereotypical movements,spontaneous activity and emotional disorders in rats;the method of ELISA was used to determine the levels of DA,GABA,GLU and 5-HT in the striatum and the levels of Ang II and AVP levels in the serum.Results APO,DOI,IDPN induction moulding results show that compared with the control group,the three modeling methods could successfully induce spontaneous activity,stereotypical movements,cognitive and emotional disorders in rats.The IDPN group showed more pronounced stereotyped movements,cognitive impairment and a tendency to anxiety and depression,with reduced levels of DA and GLU and increased levels of 5-HT compared with the control group(P<0.05);the APO group showed more pronounced spontaneous activity and increased excitement,irritability,exploratory and even aggressive behaviour,with increased levels of DA and GLU(P<0.05)and significantly higher levels of Ang II compared to the control group(P<0.01);the behavioural performance of the rats in the DOI group was similar to that of the APO group,but the degree of spontaneous activity and stereotyped movement was worse than that in the APO group,and 5-HT levels were significantly lower in the DOI group compared with the control group(P<0.01).The results of the APO group loaded with RSS and AESS modelling respectively showed that the spontaneous activity in the APO+RSS group were increased,stereotyped movements and exploratory behaviour compared with the APO group;the APO+AESS group showed more pronounced cognitive impairment.Striatal DA,GLU levels and serum Ang II were increased in the APO+RSS group compared to the APO group and APO+AESS group(P<0.05).Conclusion APO-induced TD model has good apparent and structural validity,and to a certain extent has the characteristics of hyperactivity of liver-yang syndrome in traditional Chinese medicine;APO combined restraint and suspension stress stimulation model activates the activation of RAS system,aggravates the symptoms of hyperliver-yang and is conducive to maintaining the stability of TD model,which is a relatively ideal animal model for the combination of disease and syndrome of hyperactivity of liver yang in TD model.
作者
张心放
吴振起
赵雪
郝欧美
王子
ZHANG Xinfang;WU Zhenqi;ZHAO Xue;HAO Oumei;WANG Zi(Graduate School of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)
出处
《环球中医药》
CAS
2024年第6期1043-1049,共7页
Global Traditional Chinese Medicine
基金
沈阳市科技计划项目(21-174-9-01)。
关键词
抽动障碍
肝阳上亢证
应激
模型
大鼠
tic disorder
syndrome of hyperactive liver yang
stress
model
rat
