Analyse The Effective Drugs Of Gandou Decoction On Neurone Protection Pathways Of Wilson’s Disease Model-tx Mouse And The Therapeutic Mechanism Study

Objective:1.To analyse the substance basis(most effective drugs)of the compound traditional Chinese medicine gandoutang(GDT) on TX mice of wilson’s disease animal model on neuron protected pathways based on uniform design experimentation,and analyse the most effective compound compatibility of GDT on neuron protected pathways.2.To study the effects of serum and brain trace element especially copper and zinc on TX mice,and analyse the most effective drugs of copper metabolism pathways.3.To study the effects of neuron apoptosis of wilson’s disease model-TX mice, test the efficacy of effective drugs and analyse the mechnism of neuron protected pathway,and provide the reliable guidance for reseaching new effective compound compatibility of wilson’s disease.4.To explore the analysis method on related substance basis of the compound traditional Chinese medicine on general pharmacologic action through different pathways.Method:1.110 2-month-old TX mices were grouped for 11 groups by table U11*(116)in uniform design,selecting Dahang, Janghang,Hanglian,Zexie,Jinqiancao,Sanqi as the four factor,arranging 11 levels according to the dosage of mices 0.2ml/10g/d for 30 days.2.10 1-month-old TX mices and 10 1-month-old DL mices are selected as control group befor treating,10 2-month-old TX mices and 10 2-month-old DL mices are selected as control group by according to the NS dosage of mices 0.2ml/10g/d for 30 days.3.The protein expression of P38、ERK、Bcl-2 and Bax in the brain of TX mice of each group were determined by Westernblot.4.The serum and brain copper, zinc, iron, molybdenum on TX mice of depression were detected by inductively coupled plasma-atomic emission spectrometry.5.Investigated brain morphology of TX mice with HE Staining.Results:1.Compared with control group animals,the serum copper level decreased had no significant difference,the brain copper level increased had no significant difference too.the serum zinc level decreased had no significant difference,the brain zinc level increased had no significant difference too.The change of the serum and brain iron and molybdenum level had no significant difference.Group 2,Group 3,Group 4,Group5,Group 8 and Group 11 could increase the level of serum copper significantly.Group4,Group 9,Group 10 and Group 11 could decrease the level of brain copper.Through multiple regression analysis, the equation had significant relation with X1(Dahuang)and X2(Janghuang),but no significant relation with X3(Huanglian),X4(Zexie),X5(Jinqiancao) and X6(Sanqi).2. The protein expression of P38,ERK,Bcl-2 and Bax determined by Western-blot.The protein expression of P38 Bax increased significantly,the protein expression of ERK and Bcl-2 decreased.Group 1,Group 2,Group 3,Group 4,Group 5,Group 6,Group8,Group 9,Group 10,Group 11 could decrease the protein expression level of P38.Group4,Group 5,Group 7,Group 9,Group 11 could increase the protein expression level of ERK.Group 1,Group 3,Group 7,Group 9,Group 10,Group 11 could increase the protein expression level of Bcl-2.Group 1,Group 5,Group 6,Group 7,Group 8 could decrease the protein expression level of Bax.Through multiple regression analysis, the equation had significant relation with X2(Janghuang),X3(Huanglian),X4(Zexie) and X5(Jinqiancao),but no significant relation with X1(Dahuang)and X6(Sanqi).3. Investigated brain morphology of TX mice with HE Staining,the brain tissue could be observed neurons interstitial edema,nucleus stain and pyknosis of 1-month-old model TX mices.the brain tissue could be observed neuronal disorders, spacing increases,a lot of stained karyopyknosis neurons,and several degeneration and necrosis neuron.Compared with model group animals,the neuronal injury decreased by treating for GDT.Conclusions:1.GDT could intervene the copper metabolism of TX mice,the probable effective drugs on the pathway was Dahuang and Jianghuang,,the best compound compatibility was Dahuang 15 g and Jianghuang 15 g.2.GDT could intervene the neuron apoptosis of TX mice.The antiapoptosis mechanism of GDT maybe menifested by multiple pathways and targets, including down-regulat the expression of P38 for reducing neuronal injury,up-regulat the expression of ERK for neuronal cytothesis,inhibited neuron apoptosis by regulating the expression of Bcl-2 and Bax.The probable effective drugs on these pathways were Jianghuang,Huanglian,Zexie and Jinqiancao,the best compound compatibility was Jianghuang 15 g,Jinqiancao18g,Huanglian 14 g and Zexie 2g.3.Sanqi had no significant difference in changing all the test index.We could remove it from GDT if we could verify it later.