| ADHD(Attention Deficit Hyperactivity Disorder,ADHD),is also known as ADHD children.By integrating the awareness of monoamine neurotransmitters in the pathogenesis of ADHD,Sonuga-Barker recent put forward a model of the "dual pathway",which is considered better psychological - pathology - physiological model.The model emphasized the Central role of the obstacles to persistent of the initiative to the attention and impulsive behavior in ADHD.the Executive dysfunction(EDF) and the "delay aversion"(DEL) together constitute the psychological - pathologic basis of ADHD.In the pathophysiology, EDF is due to disorder of the cortex beam(MC) and the substantia nigra - Striatum beam (NS) which are related branches of DA,controlling the prefrontal cortex - the dorsal striatum DA loop in the brain.The location of the main pathological is Prefrontal cortex and dorsal striatum;and DEL is due to changes in function of Mesolimbic beam which is related branch of DA,regulating and controlling prefrontal cortex - the ventral striatum of the "reward" loop,the main pathological location are the prefrontal cortex,ventral striatum and nucleus accumbens.Prevalence rate of ADHD is High,against is large.At present,central stimulants is the preferred treatment of ADHD in Western medicine,although it may control attention deficit. hyperactivity,impulsive symptoms on certain degree in the short or medium term,but more adverse reactions appear,and the effective dose have greater individual differences,after stopping taking drug,rebound quickly,Relapse soon;as controlled drugs,the potential addiction to the drug make both doctors and patients have many concerns.Over the past two decades,Chinese medicine is applied to treat ADHD in Chinese medicine practitioners in Chinese.The majority of scholars believe that the pathogenesis of this disease is an imbalance of yin and yang organs,deficiency of yin causes the movement of yang,whose relationship with the heart,liver,spleen,kidney is the most closely.Several reports have affirmed the effectiveness of the traditional Chinese medicine in the treatment of ADHD and found that its effectiveness is similar to Central stimulants,although its effectiveness is slow onset,but effect is positive long-term,and did not find obvious Side effects,less rebound nor addiction.Although each used different prescriptions,we analysis lots of reports and found that Kongsheng Zhen zhong pill is the the basic Prescription in treatment of ADHD.The Prescription is created by Simiao Sun in the Tang Dynasty.effect is to tonify kidney and heart,calm the mind.nourish spirit..Due to restrictions on technology and ethics,it is very difficult to research the mechanism of the efficacy of the human body.the experiment on animals has become an important alternative.Chinese scholars have a preliminary exploration in this regard,founding that traditional Chinese medicine treatment can reduce the autonomy of activities of laboratory animals,improving their learning and memory impairment,but these methods have an obvious flaw:animal model lack of neuropathic clear positioning,although content of the central DA and its metabolites of the experimental animals were detected,the lack of guidance of reasonable neuropsychological - pathology - physical model,materials of the experiment came from the whole brain tissue,the role of DA is not the same in different central parts,the result of Chinese medicine is difficult to determine its specific mechanism of ADHD.Obviously, this defect severely limits the study of animal experiments,the effect related to it will be also difficult to carry out experiments,greatly influenced the development of effective drugs and the clinical application,and research in ADHD need to be a breakthrough on key issues and solutions.Objective:This project chooses the classic Chinese medicine prescription on treating ADHD——Kongsheng Zhenzhong Pill and studies it's impact on the signal trasmission of the neurotransmitter DA of the prefrontal cortex-basal ganglial circuit loop in the center of the ADHD animal model——SHR.The project tries to make clear Kongsheng Zhenzhong pill's curative effect in animal experiment and explore in depth the mechanism of treating ADHD with Chinese medicine.Content and Method:The project uses the internationally accepted ADHD animal modelnowadays——spontaneous hypertensive rat.In the prefrontal cortex and lentiform nucleus, which are the mainly distributed regions of DA nerves of a rat that control the prefrontal cortex-basal ganglial circuit loop,we use immunohistochemistry and molecular biology technology to detect the influence of the gene expression and protein expression of tyrosine hydroxylase,dopamine transporter,dopamine receptor and compare them with the central stimulant methylphenidate hydrochloride.1.Use male SHR of SPF grade,which are 8-week old and weigh 240～300g,and randomly divide them into three groups.Then use Kongsheng Zhenzhong Pill, methylphenidate hydrochloride and physiological saline to caculate each rat's gavage volume(ml) in accordance with certain proportion of drug/ weight.4 weeks after gavaging, for the next experimental.2.Detect with immunohistochemistry the characteristics of the protein expression of SHR's frontal cortex,lentiform nucleusTH,DAT.DRD1,DRD2 under the effect of Kongsheng Zhenzhong Pill. 3.Detect with Western blot the characteristics of the protein expression of SHR's frontal cortex,lentiform nucleusTH,DAT,DRD1,DRD2 under the effect of Kongsheng Zhenzhong Pill.4.Use in situ hvbridization to detect the characteristics of the gene expression of SHR's lentiform nucleusTH,DAT,DRD1,DRD2 under the effect of Kongsheng Zhenzhong Pill.5.Detect with fluorescent quantitative PCR the characteristics of the gene expression of SHR's lentiform nucleusTH,DAT,DRD1,DRD2 under the effect of Kongsheng Zhenzhong Pill.Result:1.With the use of Immunohistochemistry and Western blot.the protein expression of tyrosine hydroxylase in the prefrontal cortex and lentiform nucleus of SHR from the Chinese medicine group is higher than that in the western medicine group.Meanwhile,the protein expression from the western medicine group is higher than the control group.The expression of dopamine transporter from the western medicine group is higher than that in the Chinese medicine group while both groups are lower than that in the control group.2.With the use of Immunohistochemistry and Western blot.the protein expression of dopamine receptors(including D1 and D2) in the prefrontal cortex and DRD1 in lentiform nucleus of SHR from the Kongsheng Zhenzhong Pill group is higher than that from the methylphenidate hydrochloride group while both groups are lower than the control group; the protein expression of dopamine receptors D2 in lentiform nucleus from the methylphenidate hydrochloride group is higher than that in the Kongsheng Zhenzhong Pill group and both groups are lower than that in the control group.3.The results of in situ hybridization and fluorescence quantitative PCR showed that the gene expression oftyrosine hydroxylase in the prefrontal cortex and lentiform nucleus of SHR from the Chinese medicine group is higher than that in the western medicine group. Meanwhile,the gene expression from the western medicine group is higher than the control group.The expression of dopamine transporter from the western medicine group is higher than that in the Chinese medicine group while both groups are lower than that in the control group.4.The results of in situ hybridization and fluorescence quantitative PCR showed that the gene expression of dopamine receptors(including D1 and D2) in the prefrontal cortex and DRD1 in lentiform nucleus of SHR from the Kongsheng Zhenzhong Pill group is higher than that in the methylphenidate hydrochloride group and both groups are lower than that in the control group:the gene expression of dopamine receptors D2 in the lentiform nucleus from the methylphenidate hydrochloride group is higher than that in the Kongsheng Zhenzhong Pill group while both groups are lower than that from the control group.Conclusion: 1.Kongsheng Zhenzhong Pill improves the main symptoms of ADHD animal model bv influencing the signal transmission of the DA nerves in the the prefrontal cortex-basal ganglial circuit loop of the control center.2.That Kongsheng Zhenzhong Pill can promote DA synthesis to improve the core symptoms of an animal model of ADHD is through a multi-target,multi-role approach to achieve:(1) To promote the biosynthesis of DA and peripheral presynaptic release of DA by increasing DA synthesis rate-limiting enzymes - tyrosine hydroxylase(TH) content,;(2) The impact of dopamine transporter gene expression and protein expression, thereby reducing the re-uptake of DA;(3) The impact of DA receptors(including D1 and D2) of gene expression and protein expression,increase central DA function. |
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