| Background:In recent years,the morbidity and mortality of acute myocardial infarction(AMI)have been increasing year by year in my country.Reperfusion therapy is currently the main treatment for myocardial ischemia and myocardial infarction,but there is currently a problem of myocardial ischemia-reperfusion injury(I/R).Myocardial I/R injury is the restoration of blood supply to the ischemic myocardium within a short period of time,and the formerly ischemic myocardium undergoes more severe injury than before the blood supply was restored.Exploring the pathological mechanism of myocardial I/R injury and finding protective drugs against it have become the focus of improving the clinical prognosis of AMI patients.Objectives:1 Evaluating the efficacy and safety of Chinese herbal medicine for adverse cardiovascular events and myocardial protection after percutaneous coronary intervention(PCI).2 To investigate the effects of Panax quinquefolium saponin(PQS)with Panax notoginseng saponins(PNS)on cardiomyocyte injury after I/R injury in rats,and to explore the possible mechanisms from mitophagy and apoptosis pathways.Methods:1 Literature Research SectionWe searched the full-text databases of Chinese journals(CNKI),VIP(VIP),Wanfang Data;and foreign databases including PubMed,EMBASE,and Cochrane CENTRAL by computer.we collected publicly available information on the combination of the main method of benefitting Qi and activating blood with conventional Western medicine for the treatment of coronary artery disease in patients with acute coronary syndrome PCI patients in randomized controlled clinical trials(RCTs).2 Experimental Research SectionSeventy-five healthy adult male SD rats were randomly divided into five groups:sham-operated(Sham)group,model(I/R)group,I/R+QN(PQS and PNS 200 mg-kg-l-d-1,gavaged for 4 weeks)group,I/R+QN+3-methyladenine(15 mg-kg-l-d-1 for 3-MA,gavaged for 4 weeks)group,and I/R+3-MA group,n=15 for each group.Except for the Sham group,the ligated myocardium turned white and caused myocardial ischemia,except for the Sham group,where the left anterior descending coronary(LAD)under the coronary artery was not ligated after opening the chest.ECG monitoring showing ST-segment elevation of 0.2 mV was considered as successful modelling,which lasted for 30 min before opening the ligature line,closing the chest and reperfusing for 120 min.Biochemical analyzer was used to measure serum lactate dehydrogenase(LDH)and serum creatine kinase MB isoenzyme(CK-MB)content;2%azole(2,3,5-Triphenyl-2H-Tetrazolium Chloride,TTC)phosphate buffer staining to measure myocardial infarction area;HE staining to observe myocardial pathological changes;in situ nick end labeling(TUNEL)to detect myocardial cell apoptosis in each group;observe autophagosomes between different groups under radio microscope and changes in mitochondrial ultrastructure;Western blot was used to detect the expression of apoptosis-related proteins.Results:1 Literature Research Section① Results of the literature search:41 studies were finally included,including 5 English-language studies and 36 Chinese-language studies,all in China;6380 cumulative cases,including 3200 cases in the treatment group and 3180 cases in the control group.② Major Adverse Cardiovascular Events(MACEs):Statistical Outcomes:Total incidence of MACEs,recurrence of angina,non-fatal myocardial infarction,arrhythmia,heart failure,repeat revascularization,cardiac death,in-stent restenosis,and other infarctions.For benign cardiovascular events,a meta-analysis of dichotomous variables was performed;the results were:total incidence of MACEs[RR=0.49,95%CI(0.41,0.59),P<0.00001,I2=0],recurrence of angina[RR=0.48,95%CI(0.32,0.72),P=0.0004,I2=0],non-fatal myocardial infarction[RR=0.58,95%CI(0.38,0.87),P=0.009,I2=0],arrhythmia[RR=0.66,95%CI(0.40,1.09),P=0.11,I2=0],heart failure[RR=0.36,95%CI(0.21,0.61),P=0.0002,I2=0],repeat revascularization[RR=0.50,95%CI(0.35,0.70),P<0.0001,I2=0],cardiac death[RR=0.33,95%CI(0.12,0.90),P=0.03,I2=0],In-stent restenosis[RR=0.46,95%CI(0.35,0.60),P<0.00001,I2=0]and other cardiovascular events[RR=1.0,95%CI(0.14,7.04),P=1]..③ Markers of myocardial injury:Statistical outcome indicators:CK-MB and cTnI,Meta-analysis of continuous variables was performed,and the results were:CK-MB[MD=-2.26,95%CI(-3.83,-0.70),P=0.005,I2=89%],cTnI[MD=-0.28,95%CI(-0.72,0.17),P=0.22,I2=99%].④ Evaluation indicators of cardiac function:Statistical outcome indicators:LVEF,NT-pro BNP and BNP.Meta-analysis of continuous variables was carried out.The results were:LVEF[MD=5.31,95%CI(3.95,6.67),P<0.00001,I2=89%],NT-pro BNP[MD=-68.69,95%CI(-119.52,-17.87),P<0.008,12=50%],BNP[MD=-16.68,95%CI(-23.46,-9.90),P<0.00001,I2=0%].⑤ Inflammatory factors:Statistical outcome indicators:Hs-CRP,IL-6 and TNF-α.Meta-analysis of continuous variables was carried out,and the results were:Hs-CRP[MD=-1.80,95%CI(-2.24,-1.35)),P<0.00001,I2=66%],IL-6[MD=-3.34,95%CI(-4.30,-2.37),P<0.00001,I2=33%],TNF-α[MD=-3.86,95%CI(-4.68,-3.04),P<0.00001,I2=0].⑥ TCM syndrome score:Meta-analysis of continuous variables for TCM syndrome score,TCM syndrome score[MD=-3.43,95%CI(-4.15,-2.72),P<0.00001,I2=12%].2 Experimental Research Section① General:During the experiment,the experimental animals were in good health,and there was no significant difference in body weight between the two groups of rats(P>0.05).② Myocardial infarction size:After 4 weeks of intragastric administration and reperfusion for 120 min,the myocardial infarction size of the I/R group was significantly increased compared with the Sham group(P<0.05).There was no significant difference in myocardial infarction size between I/R+QN group and Sham group(P>0.05).Compared with the I/R group,the myocardial infarction area in the I/R+QN group was significantly reduced(P<0.05).Compared with the I/R+QN group,the myocardial infarction area in the I/R+QN+3-MA group and the I/R+3-MA group was significantly increased(P<0.05).There was no significant difference in myocardial infarction size between I/R+QN+3-MA group and I/R+3-MA group(P>0.05).③ Serum LDH and CK-MB contents:After 4 weeks of intragastric administration and 120 min of reperfusion,the serum LDH and CK-MB contents of the I/R group were significantly higher than those of the Sham group(P<0.05).There was no significant difference in serum LDH and CK-MB levels between I/R+QN group and Sham group(P>0.05).Compared with the I/R group,the serum LDH and CK-MB levels in the I/R+QN group were significantly decreased(P<0.05).Compared with I/R+QN group,serum LDH and CK-MB levels in I/R+QN+3-MA group and I/R+3-MA group were significantly increased(P<0.05).There was no significant difference in serum LDH content between I/R+QN+3-MA group and I/R+3-MA group(P>0.05).④ Pathological morphology of myocardial tissue:myocardial fibers in the Sham group were neatly arranged,with clear borders,no swelling or disorder,and no inflammatory cell infiltration in the myocardial interstitium;myocardial cells in the I/R group were enlarged,the myocardial fibers were arranged in disorder,and the gap was swollen,widened,and myocardial fiber defect;myocardial fibers in I/R+QN group were basically complete,with a small amount of interstitial edema and swelling,and cell structure was relatively complete;I/R+QN+3-MA and I/R+3-MA.Cardiomyocytes in the MA group were swollen,and myocardial fibers were ruptured and defected,similar to the I/R group.⑤ Cardiomyocyte apoptosis and related molecular pathways.(1)Apoptosis index:compared with the Sham group,the number of apoptotic cells in rat cardiomyocytes was significantly increased in the I/R group(P<0.05).no significant apoptosis was observed in the I/R+QN group,and there was no significant difference compared with the Sham group(P>0.05);compared with the I/R group,the number of apoptotic cells in the I/R+QN group was significantly decreased(P<0.05).no significant difference was observed in the I/R group,the I/R+QN+3-MA group and the I/R+3-MA group(P>0.05)./R+QN+3-MA group and I/R+3-MA group showed no significant difference in cardiomyocytes(P>0.05).(2)Apoptosis-related protein expression levels:compared with the Sham group at 120 min of reperfusion,myocardial Bcl-2 protein expression was significantly decreased and cleaved caspase-3,Cyt-c and Bax protein expression levels were significantly increased in the I/R group compared with the Sham group(all P<0.05);myocardial Bcl-2,cleaved caspase-3,Cyt-c and Bax were significantly increased in the I/R+QN group(all P<0.05);myocardial Bcl-2,cleaved caspase-3,Cyt-c,and Bax protein expression levels were not significantly different in the I/R+QN group compared with the Sham group(all P>0.05);compared with the I/R group,myocardial Bcl-2 protein expression was significantly increased and cleaved caspase-3,Cyt-c,and Bax protein expression levels were significantly decreased in the I/R+QN group(P<0.05);the I/R group,I/R+QN+3-MA-group and I/R+3-MA group showed no significant difference in myocardial Bcl-2,cleaved caspase-3,Cyt-c,and Bax protein expression levels(P>0.05)⑥Cardiomyocyte autophagy and related molecular pathways.(1)Mitochondrial ultrastructure and autophagosomes in cardiomyocytes:compared with the Sham group,the mitochondrial structure of the I/R group was severely damaged,mitochondria were swollen and sparse,and some mitochondria were ruptured,showing a small number of autophagic vesicles;the mitochondrial structure of the I/R+QN group was significantly better than that of the I/R group,the mitochondrial membrane structure was basically intact,the outer mitochondrial membrane was slightly blurred,the barren structure of the inner membrane was still clear,the arrangement was slightly sparse,and the mitochondria The mitochondrial morphological structure of I/+QN+3-MA group and I/R+3-MA group showed that cardiomyocytes were obviously swollen,structurally disorganized and ruptured,as well as obviously edematous,and the number of autophagic vesicles was significantly reduced compared with that of I/R+QN group.(2)Expression levels of autophagy-related proteins:at 120 min of reperfusion,myocardial Beclinl,LC3 Ⅰ/LC3 Ⅱ protein expression ratio,BNIP3 and HIF-1α protein expression levels were significantly increased in the I/R group compared with the Sham group(P<0.05).Compared with the I/R group,myocardial Beclinl,LC3 I/LC3 Ⅱ protein expression ratio,BNIP3 and HIF-la protein expression levels were significantly increased in the I/R+QN group(P<0.05).Compared with the I/R+QN group,myocardial Beclinl,LC3Ⅰ/LC3 Ⅱ protein expression ratio,BNIP3 and HIF-1α protein expression levels were significantly decreased in the I/R+QN+3-MA group and I/R+3-MA group(P<0.05).⑦Molecular docking:Ginsenoside Re docked with HIF-1α,the results showed that the docking affinity was-7.5 kcal/mol,and the docking affinity between Ginsenoside Re and BNIP3 was-6.0 kcal/mol;Ginsenoside Rb1 was docked with HIF-lα,and the results showed that the docking affinity was-The docking affinity between Ginsenoside Rb1 and BNIP3 was-6.1 kcal/mol;the docking affinity between Ginsenoside Rg1 and HIF-1α was-6.9 kcal/mol,and the docking affinity between Ginsenoside Rgl and BNIP3 was-6.4 kcal/mol.mol;Notoinsenoside R1 docked with HIF-1α,the results showed that the docking affinity was-7.7 kcal/mol,and the docking affinity of Notoinsenoside R1 and BNIP3 was-6.5 kcal/mol;Ginsenoside Rd docked with HIF-1α,and the results showed that the docking affinity was-8.2 kcal/mol,the docking affinity of Ginsenoside Rd to BNIP3 is-7.0 kcal/mol.Conclusions:1 In the part of literature research,it is suggested that traditional Chinese medici ne for nourishing qi and activating blood circulation combined with conventional weste rn medicine can reduce the incidence of MACEs events in patients after PCI,reduce t he level of myocardial injury markers,and improve cardiac function and TCM syndro me scores.It is recommended to be popularized and applied in clinical practice.2 In the basic experiment part,it was confirmed that the combination of nourishing qi and activating blood(PQS+PNS)may inhibit myocardial cell apoptosis,protect mitochondrial structure,and reduce myocardial I/R injury in rats through the HIF-1α/BNIP3 mitophagy pathway. |
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