The Effect Of GDT On Lung Function And Immune Index Of Wilson’s Disease And Study Of Pulmonary Pathology Ultrastructure On Model Rat

Objective:(1) To explore whether there is lung function damage and dysimmunity in WD patientsand the relationship in between by observing of lung function and immune index(T-lymphocyte、immunoglobulin and C3、C4).(2) Using different treatment programmes to clinical study of Wilson’s disease,observing the clinical curative effect and that to impact on pulmonary function andimmune index,to confirm whether GDT can improve the above indexes.(3) Using copper load model in rats and drug intervention, compare of their differencesof immune indices and changes of lung tissue copper contents,observe of pathologicalchanges of lung tissue and its copper-particle deposition, thus providing an objectivebasis for clinical.Methods:(1) The67patients were selected to divide into2groups with35in the GDT+DMPSgroup and32in the DMPS group,6-day course, intermittently for2days, a total of4courses,compared with20cases of healthy group.To observe the results on before andafter therapy, which including Clinical effects、pulmonary function、T-lymphocyte、immunoglobulin、complement （C3、C4）、serum ceruloplasmin、serum copperoxidase、serum copper、24h urine copper tests、 determination of security index andanalyze the correlation between lung function and immune function.(2) By feeding containing copper sulfate to make model of rats, put them to death,getsample abdominal aortic blood and lung tissue specimens after treatment.Then useNAVIOS-flow cytometry to test T-lymphocyte, inductively coupled plasma massspectrometer to measure the copper content in lung tissue, and adopt H-E、dithiooxamide copper staining under light microscopy and electron microscopy toobserve the pathological changes of lung tissue.(3)Adopting SPSS17.0software to carry out the data asx±s and WPS Office2012software to draw statistical chart,comparison between group using t test, hierarchical information using Ridit analysis, count data using the x2test.Data were consideredsignificant when p≤0.05or p≤0.01.Results:(1)Lung function and immune function on WD patients:Compared with healthy group,the level of FEV1.0/FVC、FVC、MVV、V50、V25were significantly reduced (p<0.01)and with the duration extension, the reduce was more obvious(p <0.05or p <0.01).The immune indexes of CD3+CD4+/CD3+CD8+、CD3+CD4+weredecreased significantly (p <0.01), while CD3and CD3+CD8+had no significantdifference (p>0.05).The level of IgG and IgM in the humoral immunity indices weresignificantly higher (p<0.05or p<0.01)and complement C3, C4was significantlyreduced (p<0.01), while IgA had no significant difference(p>0.05).FEV1.0/FVC%was positively correlated with CD4、complement C4(p <0.05or p <0.01), andnegatively correlated with IgG、IgM、 IgA (p<0.05).V50was negatively correlatedwith IgG (p <0.01), positively correlated with complement C4(p <0.05). V25waspositively correlated with CD4(p <0.05).(2) Clinical studies shown:After treatment, the total efficiency rate of the treatmentgroup was94.28%,better than the control group (p <0.05). TCM syndrome integralwas significantly improved (p <0.01) in the treatment group, while the control groupwas no statistical significance (p>0.05).The value of FVC、V50、V25were signifi-cantly increased (p <0.01) in both two groups and the improvement of FVC、V25inthe treatment group was better than that of control group (p <0.05).In treatmentgroup,the value of CD3+CD4+、CD3+CD4+/CD3+CD8+were significantly increasedthan before treatment and IgG significantly was lower, C3C4significantly higher (p<0.01or p<0.05), while in control group the changes was no statistical significance (p>0.05).In both two group,the level of serum ceruloplasmin、serum copper oxidase and serumcopper had no significantly change (p>0.05).The24-hour urinary copper excretion wereboth increased in the two groups（p<0.01）and the treatment group was better than thatof the control group (p<0.05). (3) Safety index：The blood routine index of the control group after treatment wasdecreased than that before the treatment (p<0.01), while the treatment group had nosignificant difference (p>0.05) between pre treatment and post treatment.The level ofALT and AST in the treatment groups were improved after the treatment (p <0.01)andthe differences between two groups had statistically significant (p <0.01). The level ofkidney and electrocardiogram on both two groups had no significant change (p>0.05).Side effects: the rate of side effects in treatment group was11.43%,28.13%in controlgroup and there was significant difference between two groups (p <0.01).(4)Animal studies shown: Compared with the blank group, the value of CD3+CD4+inWD model rats was decreased significantly (p<0.01), CD3and CD3+CD8+had nosignificant change (p>0.05), and the pulmonary copper was increased significantly(p<0.01).Compared with the model group,CD3was decreased significantly (p <0.05),the change of CD3+CD8+and CD3+CD4+had no statistical significance (p>0.05)inPCM group;and in GTD group,the value of CD3+CD4+increased significantly (p <0.01), CD3、CD3+CD8+had no significant change(p>0.05);the lung copper in bothtwo groups had dropped (p <0.01).Compared with the PCM group, CD3+CD4+andCD3was significantly increased in GDT group(p <0.01or p <0.05)(5)Under naked eye,the lung atrophy in model rat can be seen;Under lightmicroscope,inflammatory cell infiltrate obviously, dominated by lymphocytes;Underdithiooxamide copper staining,there were sporadic distributed yellow-greengranules;Under the electron microscope,the increased lamellar vesicles、chondriosomeswell and ridge disappearance and part of collagen fibers could be observed.Thesechanges was most obvious in model group and the PCM group and the GDT group hadimproved a little.Conclusion:(1) A degree of obstructive ventilatory dysfunction and small airway damage ispresented in WD patients,and with the duration extension, the damage was moreobvious. (2) Immunologic derangement of cell immunity and humoral immune function ispresented in WD patients,and lung function with that had a sure correlation.(3) The copper overloaded model can be used to study the pathogenesis of tissuedamage caused by excessive copper accumulation and the observation of drug efficacy.(4) Copper deposit in model rat’s lung tissue, and cause to change the mitochondria,increase the lamellar vesicles and then lead to lung function and immune functionchange.(5) GDT can better improve the clinical signs of damp-heat type connotation, increasethe amount of24h urine copper, regulate immune system,improve lung function andreduce the incidence of adverse reactions in WD patients,and protect the injury lung inthe copper overloaded models.