Effect Of Yiqi Huoxue Recipe On Metabolism Of Rats With Myocardial Infarction And Its Mechanism

ObjectivesThis research was aimed to investigate the protective effect, metabolic material basis and mechanism of the supplementing qi and activating blood circulation formula on myocardium in myocardial infarction (MI) rats, by observing the formula’se protective effect, metabolic expression and mechanism on MI rats. And the change in heart structure & function, metabolism, signal pathway and organizational structure foundation had been analyzed to offer the material basis for the theory of "heart governs the blood" in Traditional Chinese Medicine.MethodsThe MI models were prepared by ligating the left coronary artery in SD rats. On the basis of the evaluation of electrocardiogram, randomly SD rats were divided into model group, the supplementing qi and activating blood group, perindopril group, Qi Shen Yi Qi pill group and rats that only were punctured but with no ligation as sham operation group, which was served as a control group. There were a total of 5 groups. Intragastric administration of corresponding drugs in each group start after the operation and 1 time per day. Then at the seventh and twenty-eighth day, observing pathological change and related cardiac function indexes of MI rats to evaluate the success of MI models and the effect of the supplementing qi and activating blood circulation formula on myocardial tissue in MI rats. Based on the NMR metabolic proteomics methods, the differences in metabolite between the supplementing qi and activating blood group and model group were figured out and the p value between the drug groups and modern group was worked out. Besides, the Akt, m-TOR and phosphorylation expression in infarction marginal zone of myocardial in rat by methods of immunohistochemistry, Western blot and immunofluorescence quantitative real-time PCR were detected to analyze related metabolism signaling pathways, which provided a basis for protective effect and the mechanism of the supplementing qi and activating blood circulation formula in MI rats.ResultsIn experiment 1, the MI model was confirmed successfully by comparing the electrocardiogram change before and after the operation between the model group and the sham operation group. After HE staining, it could be observed by ordinary light microscop e that the cardiac muscle cells in sham operation group arranged neatly, with no cardiomyocyte hypertrophy. Compared with the sham operated group, the myocardial cells ’code in the model group increased obviously, myocardial cells were in disorder, and there was inflammatory cell infiltration. And the myocardial cells’ construction and inflammatory cell infiltration were better after the intragastric administration of corresponding drugs. about the aspects of echocardiography, (1) the results in diastolic left ventricular end diastolic diameter and end systolic diameter:compared with the sham operation group, the LVIDd and LVIDs in model group rats significantly increased (P< 0.01), and after the intragastric administration of corresponding drugs, the LVIDd and LVIDs in model group rats significantly decreased (P< 0.01). (2) the results of left ventricula r volume:compared with the sham operation group, the EDV and ESV in model group rats significantly increased (P< 0.01), and after the intragastric administration of corresponding drugs,the EDV and ESV in model group rats significantly decreased (P< 0.01). (3) The results of left ventricular ejection fraction and left ventricular short axis rate:in the 28 day group, compared with the sh am operation group, the EF% and FS% in model group rats significantly increased (P< 0.01). Compared with the model group, the EF% and FS% all increased in all drug groups.In experiment 2:the differences in plasma and urine metabolism:7 days and 28 days after modeling, the metabolism in plasma and urine in the sham operation group, model group, perindopril group, Qi Shen Yi Qi pill group, supplementing qi and activating blood circulation group did not display obvious separation trend. Compared with sham operation group, there were 22 kinds of significant different metabolites in plasma and 17 in urine in modern group 7 days after modeling cooperation. Compared with sham operation group, there were 15 kinds of significant different metabolites in plasma and 24 in urine in modern group 28 days after modeling cooperation. The mainly differences of metabolites were in the metabolism of carbohydrate, lipid and amino acid. After the intervention of traditional Chinese medicine, there were significant changes in the prognosis, carbohydrate metabolism and lipid metabolism (P<0.05 or P<0.01).In experiment 3:Immunohistochemistry, Western blot and RT-PCR results of AKt, m-TOR and phosphorylation protein expression in infarction marginal zone of cardiac tissues: (1) Western blot result:compared with the sham operation group, there were significant changes in p-AKt/AKt and p-mTOR/mTOR in all modern groups at 7 and 28 day (P<0.01). Compared with the modern group, there were increased trend in p-AKt/AKt and p-mTOR/mTOR in perindopril group, Qi Shen Yi Qi pill group and supplementing qi and activating blood circulation group with significant changes at 7 and 28 day (P<0.05 or P<0.01). (2) immunohistochemical result:compared with the sham operation group, there were significant changes in p-AKt and p-mTOR in all modern groups at 7 and 28 day (P<0.05 or P<0.01). Compared with the modern group, there were increased trend in p-AKt and p-mTOR in perindopril group, Qi Shen Yi Qi pill group and supplementing qi and activating blood circulation group with significant changes at 7 and 28 day (P<0.05 or P<0.01). (3) RT-PCR result:compared with the sham operation group, there were significant changes in AKt & m-TOR m-RNA in all modern groups at 7 and 28 day (P<0.01). Compared with the modern group, there were increased expression of AKt & m-TOR m-RNA in perindopril group, Qi Shen Yi Qi pill group and supplementing qi and activating blood circulation group with significant changes at 7 and 28 day (P<0.05 or P<0.01).Conclusions1. MI model was successfully established by ligating the left coronary artery, the supplementing qi and activating blood circulation formula has certain therapeutic effect on structure and function of MI rats, such as reducing infarct size, improving heart function, reversing cell morphology and function change caused by ischemia and hypoxia. Thus it could effectively treat the myocardial dysfunction.2. The glucose metabolism, lipid metabolism, amino acid metabolism, energy metabolism and some other metabolism in MI rats changed. It showed that the supplementing qi and activating blood circulation formula had regulated effects on products of sugar, energy and lipid metabolism, such as lactic acid, glycerol, α-glucose, β-glucose, glycoprotein, fructose, nicotinic acid and choline. So it had been confirmed that there were sugar, energy and lipid metabolism disorder in MI rats’ hearts, and the supplementing qi and activating blood circulation formula had great effects.3. The supplementing qi and activating blood circulation formula could activate the AKt/m-TOR pathway to improve metabolism and thus play an important role in myocardial protection. According to the changes of cardiac function & structure, plasma and urinal metabolism and related signal pathway, this research could offer material basis for the theory of "heart governs the blood" in Traditional Chinese Medicine.