Experiment And Clinical Study On Treatment Of The Ventricular Arrhythmia In CHD With Dcordaline And The Composite Prescriptions Includining Yanhusuo

Today, coronary heart disease (CHD) has become one of the three cause of human death. The morbidity of CHD has increased year after year. Because the basic pathophysiology of CHD is myocardial ischemia, if we are going to save ischemic myocardium, we must resume the myocardial ischemic-reperfuse timely and efficiently. But the reperfusing of myocardium often causing ischemia reperfusion injury (IRI). How to control the ischemic reperfusion injury of myocardium has become an important field in basic and applying research of cardiology.Murry found that ischemic preconditioning can relieve ischemia reperfusion injury in 1986. But it has difficult application and extension in clinic because of the injury of ischemicpreconditioning. So it is of great practical significance to find safe and effective drugs for preconditioning. In 1988, the results from CAST trail suggest that: antiarrhythmic drugs have the potential danger of causing arrhythmia. This bring opportunities and challenges for traditional Chinese drugs preconditioning to control ischemia reperfusion injury, specially in antiarrhythmic induced by myocardial ischemia and reperfusion injury.Corydalis yanhusuo W. T. Wang has been used in much TCM composite prescriptions in research on preventing and curing cardiovascular disease recently, specially in ischemic heart disease and antiarrhythmic. Corydalis yanhusuo W. T. Wang, as a kind of promoting blood circulation and removing blood stasis drug, has the effect of increasing the flow of coronary, dilating vessel, improving nutritional flow in myocardium, antiarrhythmic, the modern pharmacodynamics study showed. Up to now, the experiment study of using Corydalis yanhusuo W. T. Wang preconditioning on myocardial ischemia reperfusion injury and the clinical study of using the composite prescriptions containing Corydalis yanhusuo W. T. Wang (raoren Honghua Decoction) in the treatment of ventricular arrhythmia in CHD have not been reported. This study was a beneficial evaluation in this field.OBJECTIVE: To compare the action between ischemic preconditioning and drug preconditioning on myocardial ischemia reperfusion injury by using the experimental model of rats. To evaluate the protective action of Corydalis yanhusuo W. T. Wang preconditioning on myocardial ischemia reperfusion injury in rats and the pharmacodynamics mechanism.METHODS: SD rats were randomized into six groups: sham-operation group, model group, ischemia preconditioning group, dcordaline large, moderate and low dose group. To establish rats model with myocardial ischemic-reperfuse, with the methods such as the patch clamp technique, flow cytometry, immunohistochemical technique, electron microscopic cytochemistry of enzyme, molecular biologic technique, to evaluate the protective action of dcordaline preconditioning on myocardial ischemia reperfusion injury in rats and the pharmacodynamics mechanism. This study includes 6 parts. Part 1: observation on the effect of dcordaline preconditioning on rats myocardial ischemic-reperfuse arrhythmic. Part 2: observation on the effect ofdcordaline preconditioning on myocardial ischemic-reperfuse cell apoptosis. Part 3: observation on the effect of dcordaline preconditioning on activities of Ca²⁺-ATPase and Na⁺-K⁺-ArPase in ischemic-reperfuse myocardium. Part 4: observation on the effect of dcordaline preconditioning on gating dynamics of L-type calcium currents(I_Ca-L) in single myocardial cell. Part 5: observation on the effect of dcordaline preconditioning on the concentration of free Ca²⁺ in ischemic-reperfuse myocardium. Part 6: observation on the effect of dcordaline preconditioning on the infarct size in ischemic-reperfuse myocardium.RESULTS:1. Comparion of the accident rate of ventricular arrhythmia: in dcordaline large and moderate dose groups, ventricular tachycardia (VT) and ventricular fibrillation (VF) rate were markedly reduced, there were significant differences between the tow groups and model group(P＜0.05, P＜0.01). while there were no significant differences in dcordaline low dose group(P＞0.05). There were no significant differences of the VT and VF rate between the large, moderate dose groups and model group(P＞0.05). There were no significant differences of the ventricularpremature beat (PVBs) rate among the groups(P＞0.05).2. Comparion of the occuring time of arrhythmic: in dcordaline large, moderate and low dose groups, the occuring time of arrhythmic was obviously postponed and the duration was obviously shortend, there were significant differences between these groups and model group(P＜0.01). these effect of dcordaline preconditioning were weaker than ischemic preconditioning(P＜0.01).3. Comparion of ST segment elevation level: There were no significant differences between ischemia preconditioning group, dcordaline preconditioning groups and model group(P＜0.01). Comparion of heart rate: ischemia preconditioning and dcordaline preconditioning could obviously decrease heart rate. There were significant differences between these groups and model group(P＜0.01). The effect of slowing heart rate in dcordaline Imoderate and low dose groups was weaker than that in ischemia preconditioning group(P＜0.01), while that in large dose group was stranger (P＜0.05).4. The apoptosis cells in the rats myocardial cells was counted by employing in situ end labeling technique. The results showed: the apoptosis status in the model group was higher than that in the sham-operation group (P＜0.01); the apoptosis myocardial cells and the apoptosis status in the ischemia preconditioning group and the three dcordaline preconditioning groups markedly decreased to the model group (P＜0.01); while there were not significant differences between the ischemia preconditioning group and dcordaline large dose group(P＞0.05).5. Expression of Bcl-2 proteins in myocardial cells: after ischemic-reperfuse, the expression of bcl-2 gene markedly decreased to the sham-operation group. The positive expression index was markedly lower than that in the sham-operation group (P＜0.01). After ischemia preconditioning and dcordaline preconditioning, the expression of bcl-2 markedly increased. The positive expression index in the ischemia preconditioning group and the three dcordaline preconditioning groups markedly increased to the model group (P＜0.01); while there were not significant differences between the ischemia preconditioning group and dcordaline large dose group(P＞0.05).6. Expression of Fas proteins in myocardial cells: after ischemic-reperfuse, the expression of Fas gene markedly increased to the sham-operation group. The positive expression index was markedly higher than that in the sham-operation group (P＜0.01). After ischemia preconditioning and dcordaline preconditioning, the expression of Fas markedly decreased. The positive expression index in the ischemia preconditioning group and the three dcordaline preconditioning groups markedly decreased to the model group (P＜0.01). While there were not significant differences between the ischemia preconditioning group and dcordaline large dose group(P＞0.05).7. Analyzing the activities of Na⁺-K⁺ATPase in myocardial cells: after ischemic-reperfuse, the activities of Na⁺-K⁺-ATPase in myocardial cells markedly decreased to the sham-operation group(P＜0.01). After ischemia preconditioning and dcordaline preconditioning, the activities of Na⁺-K⁺-ATPase in the ischemia preconditioning group and the dcordaline large, moderate dose groups markedly increased to the model group(P＜0.01). While there were not significant differences between the ischemia preconditioning group and dcordaline large, moderate dose groups(P＞0.05).8. Analyzing the activities of Ca²⁺-ATPase in myocardial cells: after ischemic-reperfuse, the activities of Ca²⁺-ATPase in myocardial cells markedly decreased to the sham-operation group(P＜0.01). After ischemia preconditioning and dcordaline preconditioning, the activities of Ca²⁺-ATPase in the ischemia preconditioning group and the dcordaline large dose group markedly increased to the model group(P＜0.01). While there were not significant differences between the ischemia preconditioning group and dcordaline large dose group(P＞0.05).9. Gating dynamics of L-type calcium currents(ICa-L) in single myocardial cell:(1) Open-state probability (NPo): NPO in the model group markedly increased to the sham-operation group (P＜0.01). NPO in the ischemia preconditioning group and the three dcordaline preconditioning groups markedly decreased to the model group (P＜0.01); while there were not significant differences between the ischemia preconditioning group and dcordaline large dose group(P＞0.05).(2) Open time: open time in the model group markedly decreased to the sham-operation group (P＜0.01). there were not significant differences between the ischemia preconditioning group, the three dcordaline preconditioning groups and the model group(P＞0.05).(3) Amplitude: amplitude in the model group markedly increased to the sham-operation group (P＜0.01). Amplitude in the dcordaline moderate and low dose groups decreased to the model group (P＜0.05), while that in the dcordaline large group and ischemia preconditioning group increased (P＜0.05).10. Concentration of free Ca²⁺ in myocardial cells: Ca²⁺ fluorescence density in the model group markedly increased to the sham-operation group (P＜0.01).Ca²⁺ fluorescence density in the ischemia preconditioning group and the three dcordaline preconditioning groups markedly decreased to the model group (P＜0.01); while there were not significant differences between theischemia preconditioning group and dcordaline large dose group(P＞0.05).11. Ischemic, infarct size in myocardium:(1) Ischemic size in myocardium: Except in the dcordaline low dose group(P＞0.05), the ischemic size in myocardium in the other groups decreased to that in the model group (P＜0.01 or P＜0.05). Except in the dcordaline low dose group(P＜0.05), the ischemic size in myocardium in the dcordaline large, moderate dose groups were the same as that in the ischemia preconditioning group(P＞0.05).(2) Infarct size in myocardium: the infarct size in myocardium in ischemia preconditioning group, the three dcordaline preconditioning groups decreased to that in the model group (P＜0.05). Except in the dcordaline low dose group(P＜0.05), the infarct size in myocardium in the dcordaline large, moderate dose groups were the same as that in the ischemia preconditioning group(P＞0.05).CONCLUSION:1. To ischemia reperfusion ventricular arrhythmic in rats myocardium, dcordaline preconditioning can not only postpone the occuring time of arrhythmic, shorten the duration, but also reduce ventricular tachycardia (VT) and ventricular fibrillation (VF) rate. We conclude that dcordaline preconditioning has antiarrhythmic effect against ischemia and reperfusion in rats myocardium.2. Dcordaline preconditioning can inhibite apoptosis of myocardial cells and protect myocardial cells from ischemia and ischemia reperfusion injury by up-regulation of the expression of bcl-2 proteins and down-regulation of the expression of Fas proteins. We conclude that dcordaline preconditioning has heart protected effect by the mechanism that similar to endogenous anti-apoptosis of ischemia preconditioning.3. Dcordaline preconditioning can markedly increase the activities of Na⁺-K⁺-ATPase and Ca²⁺-ATPase in myocardial cells, which can carry Ca²⁺ out from intercellular to extracellular through the pathway as follows: Ca²⁺-ATPase in sarcoplasmic reticulum, Na+-Ca²⁺ exchange system, Ca(2+)-ATPase sarcolemma. So as to decrease the concentration of free Ca2+ and calcium overload in myocardium to protect myocardial cells.4. Dcordaline preconditioning can decrease the open-state probability and the amplitude of L-type calcium currents(ICa-L) in myocardial cell so as to block the L-type calcium currents opening, to decrease the concentration of free Ca²⁺ and calcium overload in myocardium to protect myocardial cells.5. Dcordaline preconditioning can markedly decrease infarct size in rats myocardium induced by myocardial ischemia and reperfusion injury, same as ischemia preconditioning.OBJECTIVE: To observe the clinic effect of the composite prescriptions containing Corydalis yanhusuo W. T. Wang(Taoren Honghua Decoction) to frequent ventricular premature complexes(VPC) in patients with coronary heart disease(CHD).METHODS: A prospective study was done of frequent ventricular premature complexes in 60 patients with CHD under the randomized, controlled and single-blind rules. These patients were randomized into control and experimental groups in 1:1 proportion. The patients in experimental group received basic western medical therapy for CHD combined with Taoren Honghua Decoction; while the patients in control group received only basic western medical therapy. To compare the clinic efficacy to the patients in between control and experimental groups by observing the change of ventricular premature rate and the change of traditional Chinese medicine syndrome and symptom before and after treatment.RESULTS:1. Compare of the efficacy to ventricular premature after treatment: the results of therapy were assessed as follows: the general effective rate being 85% in experimental group, while 75% in control group. There was not significant differences between the two groups(P＞0.05). It is illustrate that the two groups has same effect.2. Compare of the 24h ventricular premature beats pre-treatment and post-treatment: A design of oneself comparison according to pre-treatment and post-treatment was performed in both groups. There was significant differences in each group(P＜0.01). The value between pre-treatment and post-treatment in experimental group had no difference to that in control group(P＞0.05).3. Compare of the efficacy to traditional Chinese medicine syndrome between control and experimental group: the general effective rate being 90% in experimental group, while 70% in control group. There were significant differences between the two groups(P＜0.05).4. Compare of the scores of traditional Chinese clinical symptoms between control and experimental group: A design of oneself comparison according to pre-treatment and post-treatment was performed in both groups. There were significant differences between pre-treatment and post-treatment in each group(P＜0.05). The value between pre-treatment and post-treatment in experimental group had significant difference to that in control group(P＜0.05).5. Compare of the traditional Chinese medicine symptom between control and experimental group: the general effective rate to palpitation being 89.7% in experimental group, while 66.7% in control group. There were significant differences between the two groups(P＜0.05). The general effective rate to chest stifling being 76.7% in experimental group, while 45.8% in control group. There were significant differences between the two groups(P＜0.05). There were not significant differences of other symptom between the two groups(P＜0.05)CONCLUSION:Though the observation of treatment of the composite prescriptions containing Corydalis yanhusuo W. T. Wang(Taoren Honghua Decoction) for frequent ventricular premature complexes in 30 patients with coronary heart disease(CHD), we found that the basic western medical therapy for CHD combined with Taoren Honghua Decoction has the same efficacy as single western medical therapy in decreasing frequent ventricular premature complexes and antiarrhythmic in patients with coronary heart disease. But the improvement of traditional Chinese medicine syndrome and symptom, specially palpitation and chest stifling, is significantly increased in experimental group than in contro group, we also found that the basic western medical therapy combined with Taoren Honghua Decoction has lower side effect, little causing arrhythmia.CONCLUDING:1. To ischemia reperfusion ventricular arrhythmic in rats myocardium, dcordaline preconditioning can not only postpone the occuring time of ventricular arrhythmic, shorten the duration, but also reduce ventricular tachycardia (VT) and ventricular fibrillation (VF) rate. The results show that dcordaline preconditioning has antiarrhythmic effect against ischemia and reperfusion in rats myocardium2. Dcordaline preconditioning can inhibite apoptosis of myocardial cells and protect myocardial cells from ischemia and ischemia reperfusion injury by up-regulation of the expression of bcl-2 proteins and down-regulation of the expression of Fas proteins. The results show that dcordaline preconditioning has heart protected effect by the mechanism that similar to endogenous anti-apoptosis of ischemia preconditioning.3. Dcordaline preconditioning can markedly increase the activities of Na+-K+-ATPase and Ca²⁺-ATPase in myocardial cells, which can carry Ca²⁺ out from intercellular to extracellular through the pathway as follows: Ca²⁺-ATPase in sarcoplasmic reticulum, Na+-Ca²⁺ exchange system, Ca²⁺-ATPase sarcolemma. So as to decrease the concentration of free Ca2+ and calcium overload in myocardium to protect myocardial cells.4. Dcordaline preconditioning can decrease the open-state probability and the amplitude of L-type calcium currents(ICa-L) in myocardial cell so as to block the L-type calcium currents opening, to decrease the concentration of free Ca²⁺ and calcium overload in myocardium to protect myocardial cells.5. Dcordaline preconditioning can markedly decrease infarct size in rats myocardium induced by myocardial ischemia and reperfusion injury, same as ischemia preconditioning.6. Though the clinic observation we conclude that the basic western medical therapy for CHD combined with Taoren Honghua Decoction has the same efficacy as single western medical therapy in decreasing frequent ventricular premature complexes and antiarrhythmic in patients with coronary heart disease. But the improvement of traditional Chinese medicine syndrome and symptom, specially palpitation and chest stifling, is significantly increased in experimental group than in contro group. The combined therapy has lower side effect, little causing arrhythmia, so as to has a good prospect of application and extension...