| Myocardial ischemia is the main pathological change of coronary heart disease.The decrease of blood perfusion makes the myocardium in the state of ischemia and hypoxia,resulting in abnormal energy metabolism and different degrees of reduction of cardiac function.In severe cases,it can induce adverse events such as myocardial infarction and sudden death.Timely reperfusion therapy can restore blood supply in a short time,but it will bring serious reperfusion injury while revascularization,and irreversible damage to myocardial structure and function,which was regarded as the main adverse factor affecting its prognosis.Professor Cao Hong-xin proposed "Wen yang yi xin" therapy for treating coronary heart disease based on its pathological characteristics of "Heart yang deficiency,Phlegm and blood stasis",and created the representative prescription "Wenxinfang",the preliminary research showed that it has a protective effect on myocardial ischemia and ischemia-reperfusion injury.At present,a consensus has been reached that the impairment of mitochondrial function and energy metabolism are one of the mechanisms of myocardial ischemia-reperfusion injury.Therefore,this study discussed the possible mechanism of " Wen yang yi xin" therapy in the prevention and treatment of myocardial ischemia-reperfusion injury from the perspective of mitochondrial energy metabolism.Objective:1.To study the effect of“Wen yang yi xin" therapy on the energy metabolism of myocardial mitochondria after myocardial ischemia-reperfusion injury.2.Deeply study the molecular mechanism of " Wen yang yi xin" therapy on improving myocardial mitochondrial energy metabolism,and reveal the effect of "Wen yang yi xin" therapy on AMPK/SIRT1-PGC-lα-NRF-1/ERRs energy signal axis,to provide more scientific basis for the prevention and treatment of myocardial ischemiareperfusion injury.Methods:90 male Wistar rats of SPF grade were randomly divided into sham operation group,model group and Wenxin Formula low,medium and high dose groups,with 18 rats in each group,which were given 0.99g·kg-1,1.98g · kg-1 and 3.96g·kg1 granules by gavage respectively,the sham operation group and model group were given the same amount of normal saline by gavage.21 days after pre administration,the rat model of myocardial ischemia-reperfusion injury(MIRI)was established by ligation of the left anterior descending coronary artery for 30 min and reperfusion for 2 h,whereas in the sham operation group were only threaded without deligation.After reperfusion,blood was taken from the abdominal aorta,the heart was quickly removed,and the myocardial tissue in the ischemic area of the left ventricle of the heart was cut off,extraction of myocardial mitochondria by differential centrifugation.Myocardial infarction area was observed by TTC staining,the pathological morphology of myocardial tissue observed through the Hematoxylin Eosin(HE)staining.The ultrastructure of myocardium and mitochondria were observed by transmission electron microscope.Using ELISA methods to detect the Serum CK-MB activity,using micro methods to detect the activities of LDH,ATP,PDH,α-KGDH,SDH and CCO,WIS-8 methods to detect the content of NADH.RT-PCR was used to detect the mRNA expression of AMPK,SIRT1,PGC-1α,ERRα,ERRγ,NRF-1 and TFAM in myocardial tissue,immunohistochemistry was used to detect PGC-1α Protein expression,western blot was used to detect the AMPK,p AMPK,SIRT 1,PGC-1α,ERRα,NRF-1 and TFAM expression.Results:1.The myocardial infarction area of Wenxin Decoction pretreatment group decreased,and there was significant difference in high-dose group(P<0.01),the pathological morphology of myocardial tissue was improved,the activities of serum CK-MB,LDH and tissue LDH decreased significantly(P<0.05),which alleviated myocardial ischemia-reperfusion injury.2.The ultrastructural damage of myocardial mitochondria in the model group was lighter than that in the model group,ATP content in serum and tissue,the activities of LDH,ATP,PDH,α-KGDH,NADH,SDH and CCO were increased,which of the middle and high dose groups were the most obvious or significant(P<0.05,P<0.01).3,AMPK,SIRT1,PGC-1α,ERRα,ERRy,NRF-l and TFAM mRNA were up-regulated(P<0.05,P<0.01),the positive expression of PGC-1α was more obvious(P<0.05),the phosphorylation level of AMPK protein increased,the expression of SIRT1,PGC-lα,ERRα,NRF-1 and TFAM was increased in varying degrees(P<0.05).Conclusions:1."Wen yang yi xin" therapy can protect myocardial ischemiareperfusion injury.2." Wen yang yi xin" therapy can protect the structure and function of mitochondria,improve the efficiency of tricarboxylic acid cycle and mitochondrial oxidative phosphorylation,through which to improve the energy metabolism of myocardial mitochondria after myocardial ischemia-reperfusion injury.3."Wen yang yi xin" therapy can activate "AMPK/SIRT1-PGC-1α-NRF-1/ERRs "energy signal axis,promote mitochondrial transcription and biogenesis,accelerate the efficiency of oxidative phosphorylation,so as to improve mitochondrial energy metabolism after myocardial ischemia-reperfusion injury. |
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