A Study On Screening Of The Effective Drugs And Clinical Curative Effect Of Gandou Decoction Intervention In Renal Damage In Wilson Disease

Objective 1.To screen out the effective effective drug and analyse the most effective compound compatibility in components in compound Gandou Decoction,which are responsible for intervening in renal damage in WD based on a uniform design,Aimed at optimizing dosage-regimen;2.To explore the mechanism of intervention in renal damage in WD for GDT;3.To verify the clinical curative effect for GDT intervention in renal damage in WD in a clinical trial.Methods In the animal experiment: 1.A U11*(116)dosage-regimen with the six flavors drug factors,including: Rhizoma Curcumae Longae,Coptis Chinensis,Alisma Orientalis,Lysimachia Christinae,Radix Notoginseng,and Rheum Officinale based on the uniform design used in this study were worked out;2.110 2-month-old TX mice were grouped into 11 treatment-groups,which were treated by the 11-levels formulas decoction base on table U11*(116)in uniform design in a dosage of mices 0.2ml/10g/d using intragastric administration for 30 days,and the kidneys in mice were taken out;3.10 2-month-old TX mice and 10 2-month-old DL mice are separately selected as the model group or the control group,treated by purified water in a dosage the same with the treatment-groups using intragastric administration for 30 days,the kidneys in mice were taken out;4.The concentration of the microelement Cu?Fe?Zn in the nephrocyte of mice each group were detected using atomic absorption method;5.Investigating kidney tissue morphology of mice each group using HE Staining;6.The expression of Bax?Bcl-2?caspase-3??-SMA and CIV in the nephrocyte of mice each group mice were detected using Western blot.In the clinical experiment: 1.All patients with WD for the first time in hospital filtered under the set of conditions,were randomly divided into the experimental group treated with Metal complexing agent DMPS combined Gandou tablets and the control group treated only use Metal complexing agent DMPS according to admission number;2.Both the serum specimens and urine specimens were taken from the two groups respectively before decoppering treatment,at the time of decoppering treatment for 4 courses,at the time of decoppering treatment for 8 courses;3.The content of the biomarkers in early stage of renal damage such as urine NAG?urine RBP?urine ?2-MG?urine m AIb?serum Cysc?urine PC??urine CIV and 24 h urine copper in the specimens were detected respectively.Results In the animal experiment: 1.Compared with control group mice:the content level of Cu(P<0.01)and Fe(P<0.01)in the nephrocyte of TX mice of 3 months had a significant increase,and the content level of Zn in the nephrocyte of TX mice of 3 months had a significant decrease(P<0.01).By the multivariate regression equation:the effect drugs screened out in GDT related to the content of Cu and Zn in the nephrocyte of TX mice were participation notoginseng(the maximum dose in uniform design 0.009 g should be taken)(P=0.030<0.05)?Rhizoma Curcumae Longae(the maximum dose in uniform design 0.059 g should be taken)(P=0.003<0.01),and no effect drug was screened out in GDT related to the content of Fe in the nephrocyte of TX mice;2.Compared with control group mice: the expression level of the protein Bax(P<0.01)?Caspase-3(P<0.05)related to apoptosis in the nephrocyte of TX mice of 3 months had a significant increase,and the expression level of the protein Bcl-2(P<0.01)had a significant decrease;the expression level of the protein SMA(P<0.01)?CIV(P<0.01)related to fibrosis had a significant increase;By the multivariate regression equation: the effect drugs screened out in GDT related to the apoptosis protein Bax?Bcl-2?Caspase-3 in the nephrocyte of TX mice were Coptis Chinensis(the maximum dose in uniform design 0.059 g should be taken)(P=0.008<0.01)?Alisma Orientalis(the maximum dose in uniform design 0.072 g should be taken)(P=0.014<0.05)?Lysimachia Christinae(the maximum dose in uniform design 0.072 g should be taken)(P=0.008<0.01)?Radix Notoginseng(the maximum dose in uniform design 0.009 g should be taken)(P=0.014<0.05);and the effect drugs screened out in GDT related to the fibrosis protein ?-SMA?CIV in the nephrocyte of TX mice were Rheum Officinale(the maximum dose in uniform design 0.059 g should be taken)(P=0.039<0.05)?Coptis Chinensis(the maximum dose in uniform design 0.059 g should be taken)(P=0.033<0.05)?Alisma Orientalis(the maximum dose in uniform design 0.072 g should be taken)(P=0.007<0.01);3.Investigating kidney tissue morphology of mice each group using HE Staining: Under a Optical microscope,compared with the control group mice,pathological changes such as glomerular swelling,great quantities of inflammatory cell infiltration into renal interstitial,renal tubular expansion in the nephrocyte of 3-month-model group mice could be observed.Compared with the model group mice,the pathological changes in 11 treatment-groups alleviated after treating for GDT in different degrees,especially in the TX mice of formula 9 group and formula11 group.In the clinical experiment: 1.There was no statistical differences in the level of 24 h urine copper(P= 0.606>0.05)?urine NAG(P=0.883>0.05)?urine RBP(P=0.981>0.05)?urine ?2-MG(P=0.980>0.05)? urine m AIb(P=0.765>0.05)? serum Cysc(P=0.675>0.05)?urine PC?(P=0.436>0.05)?urine CIV(P=0.841>0.05)between the control group and the experimental group prior treatment.2.After treating,compared with the control group patients,the level of urinary copper excretion had a significant increase(P=0.001<0.01),and the level of early renal damage such as urine RBP(P=0.008<0.01)?urine ?2-MG(P=0.000<0.01)?serum Cysc(P=0.001<0.01)?urine CIV(P=0.000<0.01)had a significant decrease in the experimental group patients.3.After treating,there was no statistical differences in the level of urine NAG(P=0.054>0.05)?urine m AIb(P= 0.152>0.05)?urine PC?(P=0.129>0.05)between the two groups.Conclusion 1.The best compatibility of GDT responsible for protection of nephrocyte in WD was screened in this animal experiment was Rhizoma Curcumae Longae 0.059 g,Coptis Chinensis 0.059 g,Rheum Officinale 0.059 g Alisma Orientalis 0.072 g,Lysimachia Christinae 0.072 g,Radix Notoginseng 0.009 g,what correspond to the original compatibility of GDT.To a certain extent,the rationality of original compatibility of GDT was confirmed.2.In the clinical experiment,it showed that by treating for GDT,the level of urinary copper excretion could be increased and the level of early renal damage biomarkers such as urine RBP?urine ?2-MG?serum Cysc?urine CIV could be decreased.The results showed that GDT play a role intervening in renal damage in WD by increassing the level of urinary copper excretion,at the same time improving the glomerular filtration and renal tubular reabsorption,and delaying the process of renal interstitial fibrosisIt.