| BackgroundReperfusion therapy of acute myocardial infarction (AMI) is a milestone in the treatment of coronary heart disease, which are mainly three ways:intravenous thrombolysis, percutaneous transluminal coronary angioplasty and stent implantation (PTCA+PCI) and coronary artery bypass grafting (CABG). With the development of interventional devices and techniques, PCI has become the main means of treatment of coronary heart disease with AMI.However, Reperfusion therapy can not stop the pathological process of coronary heart disease because it just only a kind of focal treatment measure.A series of myocardial ischemia reperfusion therapy after reperfusion injury (MIRI) severely restricted the coronary revascularization treatment effect. Inflammation, oxidative damage, calcium overload and energy utilization in disorder play an important role in the process of MIRI. in which calcium overload is a common pathway of myocardial cell injury and death. Therefore, prevention and treatment of calcium overload is an important measure to reduce MIRI injury. At present, the western medicine intervention for MIRI is limited to the experimental study, a lot of drugs have not yet been applied to clinical. The experimental study of calcium overload focus on the treatment of other organs (such as liver and ovary) after reperfusion with amlodipine.But. amlodipine acting on vascular smooth muscle of the L-type calcium channel to inhibit calcium overload, which have limited scope of application in the clinical due to the action target single, such as which can significantly inhibit cardiomyocyte contractility. Some traditional Chinese herbs also has a similar calcium antagonism, also its function characteristics and mechanismhas not been reported compare with benzene sulfonic acid amlodipine (Norvasc)--an effective western medicine calcium channel blockers in the clinical.My tutor created Yiqi Huoxue Anshen compound (inafter referred to as the compound) based on myocardial ischemia reperfusion injury in the etiology, pathogenesis and combined with years of clinical experience. It is mainly composed of Astragalus. Salvia. Radix notoginseng, Ginkgo, Spina date seed, etc. We respectively extracted the main efficacy components of those herbs, Calycosin-7-glucoside, Panax Notoginseng Saponins, Tanshinoneand salvianolic acid, Ginkgo biloba flavonoids glycol glycosides and Bilobalides,Jujuboside A and Spinosin. Therefore, aiming the two main objectives in the treatment of MIRI including calcium overbad and protection of cardiac function also inhibition of ventricular remodeling, this experimental study wasdesigned to investigate the effect of compound on myocardial injury induced by myocardial ischemia reperfusion in rats and its potential mechanisms.ObjectiveTo investigate the effect of Yiqi Huoxue compound on premature ventricular remodeling, cardiac ejection fraction and myocardial infarction area/ischemic area ratio in myocardial ischemia reperfusion injury rats.And to explore its mechanism on miRNA-133 that regulating calcium overload and cell apoptosis.Method225 male Wistar rats, body weight was 300 ± 20g, they were randomly and equally divided into 6 groups:the sham operation group, model group, ambdipine group and Yiqi Huoxue compound high, medium and low dose group.Prophylactic administration 7 days later,the left anterior descending branch of the rat heart was ligatured for 50 minutes, then wemade the model of myocardial ischemia-reperfusion injury in rats through cut the ligation line.Cardiac ejection fraction and ventricular diameter were detected before peration, after perationand and after 48h respectively. Then,the rats were killed and blood was collected, centrifuged to obtain serum, Clipping heart, assessing the myocardial infarction and ischemic area percentage by Evans blue-TTC double staining. Accord ing to those results, the optimal dosage of Yiqi Huoxue compound was chosen for the follow-upexperime ntal study to explore its mechanism.Six rats were selected in each group, and the HE staining and TUNEL staining were performed.The content of calcium ion in cardiac muscle was detected by colorimetric method, and the expressions of SERCA,CasR,MAPK,CytC,Caspase-3, Bcl-2 were determined by western blot(WB). miRNA-133 was measured by Real-Time PCR.Results(1)Cardiac ejection fraction:①After the operation:as compared with sham operation group,the EF values in the other 5 groups were significantly lowered (P<0.05),while there are no difference in the other 5 groups.②After reperfusion 48Has compared with after the operation, the EF values in model group were significantly lowered (P<0.05),the EF values in the other 5 groups were elevated (p>0.05).③After reperfusion 48Has compared with sham operation group,the EF values in the other 5 groups were significantly lowered (p<0.01).As compared with model group,the EF values inYiqi Huoxue compound high, medium and low dose group were significantly elevated (P<0.05),while there was no significantly difference between them (P>0.05)(2)LVIDd and LVIDs:①LVIDd:as compared with sham operation group, the LVIDd in model group andYiqi Huoxue compound low dose group were significantly enlarged (P<0.05).As compared with model group.the LVIDd inYiqi Huoxue compound high dose group were significantly lowered (P<0.05).②LVIDs:as compared with sham operation group the LVIDs in model group was significantly enlarged (P<0.01).As compared with model group,the LVIDs in Yiqi Huoxue compound high and medium dose group were significantly lowered (P<0.01).while there was no difference between them (p>0.05)(3)Myocardial infarction/ischemic area ratio(ranged from 30%～60%)As compared with model group.the myocardial infarction/ischemic area ratio in amlodipine group andYiqi Huoxue compound high dose group were significantly lowered (P<0.05).while there was no difference between of them (P>0.05)(4)Calcium content of the myocardium:As compared with sham operation, the calcium content of the myocardium in model group were significantly elevated(P<0.01).while amlodipine group and Yiqi Huoxue compound high dose group were significantly lowered (P<0.05).And there was no difference between amlodipine group and Yiqi Huoxue compound high dose group (P>0.05)(5)In the expression of SERCA2A and CasR:①the expression of SERCA2A:as compared with sham operation group.the expression of SERCA2A in model group were significantly lowered(P<0.01).As compared with model group,the expression of SERCA2A in amlodipine group and Yiqi Huoxue compound high dose group were significantly elevated(P<0.01),while there was no difference between them(P>0.05).②The expression of CasRas compared with sham operation group,the expression of CasR in model group, amlodipine group and Yiqi Huoxue compound high dose group were significantly elevated (P<0.01).As compared with model group,the expression of CasR in Yiqi Huoxue compound high dose group was significantly lowered (P<0.01).As compared with amlodipine group,the expression of CasR in Yiqi Huoxue compound high dose group was significantly lowered (P<0.01)(6)Myocardial apoptosis indexas compared with sham operation group, the apoptosis index in model group, amlodipine group andYiqi Huoxue compound high dose group were significantly elevated (P<0.01).As compared with model group,the apoptosis index in amlodipine group andYiqi Huoxue compound high dose group were significantly lowered (P<0.01),while there was nodifference between two of them (P>0.05).(7)In the expression of CytC,Caspase-3 and Bcl-2:①the expression of CytC:as compared with sham operation group,the expression of CytC in model group and Yiqi Huoxue compound high dose group were significantly elevated (P<0.05).As compared with model group the expression of CytC in amlodipine group and Yiqi Huoxue compound high dose group were significantly lowered (P<0.05),while there was nodifference between amlodipine group and Yiqi Huoxue compound high dose group (P>0.05).②the expression of Caspase-3:as compared with sham operation group,the expression of Caspase-3 in model group,amlodipine group and Yiqi Huoxue compound high dose group were significantly elevated (P<0.01).As compared with model group,the expression of Caspase-3 in amlodipine group and Yiqi Huoxue compound high dose group were significantly lowered (P<0.01),while there was no difference between two of them (P>0.05).③the expression of Bcl-2:as compared with sham operation group,the expression of Bcl-2 in model group andamlodipine group were significantly lowered (P<0.01).As compared with model group,the expression of Bcl-2 in amlodipine groupand Yiqi Huoxue compound high dose group were significantly elevated (P<0.01).As compared with amlodipine group,the expression of Bcl-2 in Yiqi Huoxue compound high dose group was significantly elevated (P<0.01).(8)In the expression of P38,JNKand ERK1/2:①the expression of P38:as compared with sham operation group,the expression of P38 in model group,amlodipine group andYiqi Huoxue compoundhigh dose group were significantly elevated (P<0.01).As compared with model group,the expression of P38 in amlodipine group and Yiqi Huoxue compound high dose group were significantly lowered (P<0.01),while there were no difference between two of them (P>0.05).②the expression of JNK:as compared with sham operation group.the expression of JNK in model group were significantly elevated (P<0.01), while there were no difference among of the others (P>0.05).③the expression of ERK1/2:as compared with sham operation group,the expression ofERK1/2 in model group,amlodipine group andYiqi Huoxue compound high dose group were significantly elevated (P<0.05).As compared with model group,the expression of ERK1/2 in amlodipine group were significantly lowered (P<0.01).As compared with amlodipine group,the expression of ERK1/2 in Yiqi Huoxue compound high dose group was significantly elevated (P<0.05)(9)In the expression of miRNA-133 as compared with model group.the expression of miRNA-133 in sham operation group,amlodipine group andYiqi Huoxue compound high dose group were significantly elevated (P<0.01).while there were no difference among of three of them (P>0.05)Conclusion(1)Both Yiqi Huoxue compound and amlodipine can reduce myocardial infarction/ischemic area ratioin rats when myocardial ischemia reperfusion injury occurs. In addition,Yiqi Huoxue compound can improve cardiac ejection fraction and inhibit the expansion of LVIDd and LVIDs.Thereby which can inhibit early malignant left ventricular remodeling and improve cardiac function,so its comprehensive effect is obviously better than amlodipine.(2)Yiqi Huoxue compound could inhibit calcium overload during myocardial ischemia reperfusion. and then alleviate myocardial apoptosis in the mitochondria pathway that caused by calcium overload.(3)Yiqi Huoxue compoundprotected the damaged myocardium by regulating p38MAPK pathway and up-regulating the level of miRNA-133. |
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