Effect Of DAA-â…  And Danshensu On Myocardial Apoptosis During Heart Failure Induced By Adriamycin In Rat

Objective:To observe the effect of des-asparticacid-angiotensin I(DAA-I) and Danshensu on the rats of heart failure induced by adriamycin(ADR),to explore its protective mechanisms of anti-oxidative damage and myocardial apoptosis in heart failure,to evaluate the protective effect of DAA-I and Danshensu on myocardial cells and the treatment on cardiovascular diseases,to provide the basis of pre-clinical experiments and bedding for further clinical research.We established the experimental model of heart failure in rats by intraperitoneal injection of ADR.28 days later,these rats were randomly divided into eight groups:normal control group,model group,DAA-I group,Danshensu group,DAA-I+Danshensu group,indomethacin group,DAA-I+ indomethacin group,valsartan group,at the same time it began to give intervention with drugs such as DAA-I,Danshensu,valsartan for 28 days. Hemodynamic studies such as LVSP,LVEDP,±dp/dtmax and other hemodynamic parameters were performed in each group of rat after 56 days.And then,TNF-a in plasma was detected by radioimmunoassay.SOD,MDA,CAT and other indicators of oxidative damage was detected from left ventricular tissue homogenated.Besides,pathological changes can be observed through left ventricular biopsy under a microscope.TUNEL technique can be in the charge of detecting the apoptosis of left ventricular myocardial cell.What't more,the changes in gene expression of Bcl-2,Bax,Caspase-3 were detected by immunohistochemical SABC method.Results:1.Hemodynamics:Compared with the normal group,HR and LVEDP of model group were significantly higher in rats(P＜0.01),and LVSP,±dp/dtmax were significantly lower(P＜0.01);Compared with the model group,LVEDP of the treatment groups decreased significantly(p＜0.01),and LVSP,±dp/dtmax were significantly increased(P＜0.01), which DAA-I+Danshensu group showed the most significant improvement(P＜0.01) that has not yet reached the level of normal group(P＞0.05);Compared with the model group, indomethacin group and DAA-I+indomethacin group don't have significant difference (P＞0.05). 2.Microscopic observations of the pathomorphology:clear structure,regular alignment, uniform nuclear staining had been observed in myocardial fibers of the normal rats that there was no edema,fibrosis and other obvious pathological changes;Cell degeneration such as vacuolization and edema,nuclear stained darkly,myocardial fibrosis with inflammatory cell infiltrated,atrophic myocardial fiber with disorder and widening gap between myocardial cells can be seen in the model group;Compared with model group, each treatment group has been improved,DAA-I group,DAA-I+Danshensu group,valsartan group and Danshensu group in particular.3.Cytokines changes:Compared with the normal group,the TNF-a concentration in plasma of model group was significantly higher around four times(P＜0.01);Compared with the model group,TNF-a were significantly lower(P＜0.01) in DAA-I group,DAA-I+ Danshensu group,valsartan group and Danshensu group in addition to indomethacin group,DAA-I + indomethacin group.Particularly it is most evident of DAA-I group and DAA-I + Danshensu group to decline approaching to the level of normal group(P＞0.05). Although Danshensu group had a downward trend,but there was no statistical significance(P＞0.05).4.Oxidative damage indicators:SOD,CAT in myocardial tissue of model group were significantly lower than normal group,and the content of MDA as lipid peroxidation product was higher than the normal group(P＜0.01);Compared with the model group,the treatment groups had clear differences(P＜0.01)in elevating the activity of SOD and CAT, in addition depressing MDA content;Compared with the model group,there was no significant difference in indomethacin group and DAng-I + indomethacin group(P＞0.05).5.Bcl-2,Bax,Caspase-3 gene expression:Compared with the normal group,the model group had a significantly difference(P＜0.01) that showed high expression of Bax gene,weak expression of Bcl-2 gene and decreased ratio of Bcl-2/Bax;Compared with the model group,the treatment groups had significantly differences(P＜0.01) that were inhibited in the expression of Bax gene and the promotion of Bcl-2 gene expression and increased in Bcl-2/Bax ratio,particularly DAA-I group,DAng-I+Danshensu group(P＜0.01),but there was no significant difference between the above two groups(P＞0.05);Compared with the model group,indomethacin group and DAA-I+indomethacin group had no statistical significance(P＞0.05);Compared with normal group,caspase-3 protein expression of model group was more higher(P＜0.01);Compared with the model group,the treatment groups had significantly differences(P＜0.01) that were inhibited in the expression of Caspase-3,but indomethacin group,DAA-I+indomethacin group had no statistical significance(P＞0.05).6.rate of cardiomyocyte apoptosis(AI):Compared with the normal group,the rate of cardiomyocyte apoptosis of model group that was in the position of high level had significant difference(P＜0.01);Compared with model group,cardiomyocyte apoptosis inhibited effectively had significantly difference in the treatment groups(P＜0.01), especially DAA-I group,DAA-I+Danshensu(P＜0.01),but there was no significant difference between the above two groups(P＞0.05);Compared with the model group, indomethacin group,DAA-I+indomethacin group did not significantly inhibit cardiomyocyte apoptosis(P＞0.05).7.Mortality:mortality statistics of each group in the end of the experiment showed that normal rats all survived and the treatment groups of rats with heart failure can be reduced mortality and improved the prognosis of heart failure comparing with the model group.Conclusion:1.Pathophysiological process of heart failure is closely correlated with oxidative damage and cardiomyocyte apoptosis.It is of important clinical significance to take measures of anti-oxidative damage and myocardial apoptosis on the reversal of myocardial remodeling and heart failure for avoiding or delaying the process of decompensation in time.2.DAA-I can significantly improve LVSP,LVEDP,±dp / dtmax and other hemodynamic parameters in rats with heart failure,with the remarkabe effect to maintain and improve heart function for preventing and controlling hearts failure.DAA-I could significantly reduce mortality in rats with heart failure and improve prognosis of heart failure.3.DAA-I that plays the antagonism role of angiotensinâ…¡has a myocardial protective effect by combining with the AT₁ receptor primarily.The experiment shows that it can improve cytokines,oxidative damage and myocardial apoptosis in the pathophysiology process of heart failure,and it is likely the mechanisms of protection of myocardial cells and blockade of heart failure.The effect of Danshensu alone is inferior to DAA-I or valsartan,and but DAA-I associated with Danshensu can significantly increase the combined synergies,especially in anti-oxidative damage,improving hemodynamics and so on.This shows that DAA-I associated with Danshensu from point of combined Traditional Chinese and Western Medicine for RASS-control to have some prospects for prevention and treatment of heart failure.4.The effect of DAA-I on myocardial protection such as anti-oxidative damage and cardiomyocyte apoptosis may be closely related to the following aspects:reducing the content of tumor necrosis factor in circulating,increasing activity of SOD and CAT in cardiomyocyte and lowering MDA content;inhibiting the expression of Bax gene and caspase-3 proteinin along with upregulatiing the ratio of Bcl-2/Bax.5.Compared with the current principal use of angiotensin-converting enzyme inhibitor (ACEI) and angiotensinâ…¡type 1 receptor antagonist(ARB) to inhibit the renin -angiotensin -system (RAS),DAA-I played antagonistic role of RASS system has comprehensive and precise advantages:First of all,binding with AT₁ receptor can block the bad effect of Angâ…¡combined with AT₁ receptor;Secondly,AT₂ receptor can be increased in the prostrate heart and Angâ…¡also can be increased after its effect was blocked by DAA-I,certainly Angâ…¡can exert favorable effect by binding with AT₂; Finally,it might be able to play the same role as ARB that can strengthen the combination of Angâ…¡and AT₂ to reverse the bad effect of Angâ…¡.DAA-I is a potential drug to protect myocardial cells.However,it has yet to be explored further in-depth research in detailed and comprehensive mechanism,and prevention and treatment of cardiovascular disease from the point of traditional Chinese and western medicine.