| Objective:(1)To explore whether there is heart damage in WD patients by observing blood pressure, cardiac color doppler echocardiography, brain natriuretic peptide (BNP) and homocysteine (HCY), blood lactate dehydrogenase (LDH), creatine kinase (CK), creatine kinase isoenzyme (CK-MB), cardiac troponin I and myoglobin (MYO), hypersensitive c-reactive protein (hs-CRP), heart type fatty acid binding protein (hFABP), ischemia modified albumin (IMA);(2)Through clinical observe lots of cases of Wilson disease wether have heart damages.Using different treatment programmes to clinical study of Wilson’s disease, observing the clinical curative effect and influence on heart function index.Using GDL and penicillamine on copper load model rats to intervene, compare the indicators of cardiac damage and heart tissue copper content changes, and observed the pathological morphological changes, thus providing an objective basis for clinical.Methods:(1) The 60 patients were selected to divide into 2 groups with 30 in the GDL+DMPS group and 30 in the DMPS group,6-day course, intermittently for 2 days, a total of 4 courses,compared with 20 cases of healthy group. To observe clinical efficacy of two groups patients, TCM syndrome integral, electrocardiogram, lie made a blood pressure, cardiac color doppler echocardiography, BNP, HCY, LDH, CK, CK - MB, cTn-I, MYO, hs-CRP, hFABP, IMA and safety indicators, and damage to the heart and correlation analysis.(2)Using copper load model rats, grouping after dosing abdominal aorta and blood to return heart tissue specimens.Then use enzyme-linked immunoassay kits measured copper load model of rats serum BNP, HCY, LDH, CK, CK - MB, cTn-T,cTnI,MYO, hs - CRP, hFABP, IMA, inductively coupled plasma mass spectrometer measuring heart tissue copper content, and the H-E dyeing, under electron microscopy to observe the pathological changes of heart tissue.Using western blot detection WD model rat heart tissue HO-1 protein expression.(3)Adopting SPSS 17.0 software to carry out the data as x ±s and GraphPad Prism5 software to draw statistical chart,comparison between groups by randomized block design analysis of variance comparison in the group by t test, hierarchical information using Ridit analysis, count data using the x2 test.Data were considered significant when P<0.05 or P<0.01.Results:(1) Heart damage index changes of WD patients1) Electrocardiogram (ECG):Compared with the healthy group of WD patients electrocardiogram (ECG) abnormal rate is significantly higher than the healthy group, and the difference was statistically significant (P< 0.01).2)Color doppler echocardiography: Compared with the healthy group of left atrial diameter in patients with WD group and right ventricular diameter were higher than in healthy group, but the difference had no statistical significance (P>0.05), ejection fraction value is lower than the healthy group, there was no statistically significant difference (P>0.05),and left the interior diameter value is higher than that of healthy group difference was statistically significant (P<0.05).3)Serological indicator: Compared with healthy group in patients with WD group CK, LDH, IMA, cTn - 1, hFABP, hs-CRP, HCY values were higher than in healthy group, and between the difference had statistical significance (P<0.01), CK-MB, BNP, MYO value is higher than health group but there was no statistically significant difference (P>0.05).Compared with healthy group, patients with WD CER, TYHM, serum content of Cu content significantly reduced, and between the difference had statistical significance (P< 0.01).(2) Clinical assessment: The experimental results show that after four treatment GDL+DMPS combination of clinical curative effect is superior to the simple use of DMPS.Compared with group before treatment, the treatment group after treatment of TCM syndrome integral value lower, and between the difference had statistical significance (P<0.01).There is a decrease in the control group and has statistically significant (P<0.05).Two groups after treatment, the treatment group of TCM syndrome integral value improved more significantly (P<0.01).(3) Symptomatic index evaluation of clinical1)ECG:Compared with the same group before treatment, the treatment group after treatment rate of abnormal electrocardiogram, the difference was statistically significant (P<0.05). The control group there was no statistically significant difference electrocardiogram changes (>0.05).2)Color doppler echocardiography:Compared with group before treatment, the treatment group after treatment of left atrial diameter, left inside diameter, right ventricular diameter is slightly decreased, but the difference have no sense of statistics to learn (P>0.05); The ejection fraction increases, and the difference was statistically significant (P<0.01). The control group after treatment there was no statistically significant difference each index (P>0.05). Comparison between two groups after treatment group, treatment group with right ventricular diameter and ejection fraction of improvement is better than with the control group (P<0.05).3) Heart damage related serological indexes:Compared with group before treatment, treatment group CK, CK-MB, LDH, BNP, HCY, hs-CRP, IMA, cTn-I, hFABP, MYO LDH its numerical all decreased after treatment and control group (P<0.01); Control group CK, BNP, HCY, hs-CRP, MA, IMA, cTn-I, hFABP, MYO also fell after treatment, but there was no statistical significance (P>0.05); Control group CK-MB rose slightly after treatment but there was no statistical difference learn significance (P>0.05). Comparison between two groups after treatment group, treatment group CK, CK-MB, LDH, HCY, hs-CRP, IMA, cTn-I, hFABP improvement were better than the control group (P<0.01 or P<0.05); The BNP, MYO is improved but it had no statistically significant differences (P>0.05).4)CER and TYHM, serum copper,24h urine copper:Compared with group within groups before treatment,2 groups of CER, TYHM and serum copper had no obvious difference (P>0.05); The difference of comparing between group after treatment is no statistically significant (P>0.05). Compared with the same group before treatment,2 groups of patients with WD 24h urine copper content were increased (P< 0.01); Comparison between groups after treatment, the treatment group 24h higher copper content is more obvious (P<0.01).(4) Safety index1) Blood routine:Group comparison before and after the treatment, the treatment group of RBC, WBC, PLT value slightly lower,(P>0.05); Control group of RBC, WBC, PLT value significantly decreased, and ithas statistically significant (P<0.01). Compare two groups after treatment, the WBC and PLT were higher than the control group (P<0.05); and RBC change has no statistical significance (P>0.05).2) Liver and kidney function: Compared with group before treatment, the treatment group and control group after treatment of ALT, AST, but the treatment group decreased more obviously, and the changehas statistically significant (P<0.05); Difference were observed in the two groups after treatment of BUN, Scr has no statistical significance (P>0.05). Comparison between two groups after treatment group, treatment group in the improvement of ALT, AST is superior to control group (P<0.01).3) Adverse reactions:WD patients of two groups before and after the abnormal ECG saw no obvious change. Treatment group was 6.7%, control group 30.00% incidence of adverse reactions, the distinguish statistically significant (P<0.05).(5)Animal experimental results1) Copper content of rat myocardial tissue:Compared with blank group, model group rats myocardial tissue copper content increased significantly, and there are statistically significant (p<0.01); Compared with model group, penicillamine group and hepatic bean spirit myocardial tissue are falling and copper content was statistically significant (P<0.01); Compared with penicillamine group liver beans spirit set of myocardial tissue copper content is slightly higher, the difference was statistically significant (P<0.05).2) Serological indexes of the rat:Compared with the blank group WD model group rats serum IMA, hFABP, LDH, CK, CK-MB, cTn-I, cTn-T, MYO, BNP,hs-CRP, HCY were significantly elevated (P<0.01); Compared with model group, penicillamine group, GDL group IMA, hFABP, LDH, CK, CK-MB, cTn-I, cTn-T, MYO, BNP, hs-CRP, HCY values were significantly lower,(P<0.01); Compared with penicillamine group and GDL group set of rat serumIMA, hFABP, LDH, cTn-I,cTn-T value decreased significantly (P<0.01); Compared with penicillamine group rats serum GDL group MYO,BNP, hs-CRP, HCY value is reduced; Compared with penicillamine group,GDL group rats serum CK, CK-MB value difference has no statistical significance (P>0.05).3) Rat heart tissue pathological changes:This study observed by naked eye have no obvious differences between groups in the rat heart tissue, scattered red glycine copper impregnation under the yellow particles.Visible heart tissue muscle wire fracture under electron microscopy, mitochondria vacuoles and more than the most obvious change in model group, penicillamine group and GDL group rats improved was most pronounced in model group, penicillamine group and the rat of GDL group were improved, and the improvement of GDL group is more obvious than penicillamine group. H-E staining were visible under the model group rats myocardial cell edema, local horizontal stripes blur and even disappeared, the above change model group obviously, penicillamine group and GDL group are slightly changed, improved slightly obviously GDL group.4) Testing results of Western blot experiments showed that the expression of model group rats myocardial HO-1 significantly lower than the blank group, suggests building 8 weeks after model group rats have a certain damage, myocardial and GDL and penicillamine intervention of two groups of rats myocardial were recovered, HO-1 of GDL group is improve relatively obvious.Conclusion:(1)WD patients there is a certain degree of heart damage.(2)Copper load model in the rat model can be used in the experiment to observe copper deposition in the pathogenesis of organ and tissue damage and drug curative effect observation.(3)First discovered problems under the load model in the rat heart tissue copper deposit, and lead to muscle, silk, mitochondria, the changes of ultrastructure change and heart serological indexes.(4)GDL can improve clinical symptoms in patients with phlegm and blood stasis mutual junction WD, increase the 24h urine copper content, have a certain role to protect the heart, to improve the ecg in patients with WD, and load model for copper in the rat heart tissue injury has certain protective effect. |
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