| In China,an average of one patient died from acute myocardial infarction(AMI)in less than one second.Cardiovascular disease deaths account for more than 40% of the total mortality rate in China.Thrombolytic therapy and percutaneous coronary interventions(PCI)effectively improve the survival rate of patients with AMI in the acute phase.However,after coronary recanalization,incomplete reperfusion of myocardial tissue may occur,even without reperfusion.This phenomenon is called noreflow(NR).NR can lead to insufficient local blood perfusion,left ventricular dysfunction,and even cardiac death.At present,there is no effective therapy for NR treatment in clinic,so it is significant to find drugs that can prevent and treat NR after coronary recanalization.Xinyue Capsule is an innovative traditional Chinese medicine developed based on the total saponins of American ginseng stems and leaves.It is mainly used in the treatment of angina pectoris of coronary heart disease with qi-yin deficiency.Studies have reported that Xinyue Capsule has the effect of reducing myocardial ischemiareperfusion injury,but its protective effects on NR and its mechanism have not been reported.Objective:Observing the protective effects of Xinyue capsules on myocardial ischemiareperfusion no-reflow rats,and exploring its molecular mechanism.Methods: Wistar rats were randomly divided into Sham,Model,Xinyue capsules(400,800 mg/kg)dose and alprostadil group.Sham and Model,0.5% CMC-Na 10 mL/kg was administered intragastrically(ig),Xinyue capsules 400 and 800 mg/kg dose group were given 10 mL/kg of the corresponding drug once a day by ig,for 7 days.Alprostadil group was administered intravenously(iv)2 mL/kg after ischemia and reperfusion.In this experiment,the rat left anterior descending coronary artery(LAD)was ligated for 2 hours after ischemia reperfuse for 2 hours.2 hours after reperfusion,Doppler ultrasound portable machine was used to detect the cardiac function of rats;Evans Blue,TTC,Thioflavin S staining were used to analyze the area at risk(AAR),area at infarct(AAI)and area at no-reflow(AAN)using Image-Pro Plus;CK-MB,AST,and LDH activity in rat serum were measured using biochemical and ELISA kits;HE staining reflected myocardial tissue pathological changes;White blood cells were counted in peripheral blood under an optical microscope;Biochemical kit to detect MPO activity in myocardial tissue;Immunohistochemical method to detect ICAM1 and CD62 L protein expression in rat myocardial tissue;ELISA kit to detect inflammatory cytokine levels in myocardial tissue;Western Blot to detect myocardial tissue NLRP3 inflammasome associated protein expression and its phosphorylation level.Results:1.Myocardial ischemia,infarction,and no-reflow did not occur in Sham.Compared with sham,myocardial AAR/LV,AAI/AAR and AAN/LV in Model were significantly increased(P<0.01);compared with Model,Xinyue capsules 400,800 mg /kg and positive drugs can significantly reduce myocardial AAR/LV,AAI/AAR,AAN/LV in rats(P<0.05 or P<0.01).2.Compared with Sham,LVEF and LVFS of Model rats were significantly reduced(P<0.01),while LVIDs were significantly increased(P<0.01),but no significant effect on LVIDd;compared with Model,Xinyue capsules 800 mg/kg can significantly increase LVEF and LVFS in rats(P<0.05),while Xinyue capsules 400,800 mg/kg and positive drugs can reduce LVIDs to varying degrees,but the results were not statistically significant(P>0.05).3.Compared with Sham,the serum levels of CK-MB,AST and LDH in Model were significantly increased(P<0.01);compared with Model,Xinyue capsules 400 and 800 mg/kg can reduce the activity of CK-MB,AST and LDH in rat serum(P<0.05 or P<0.01).Positive drugs can also reduce the activity of AST and LDH in serum of myocardial ischemia-reperfusion no-reflow rats(P<0.05),reduce CK-MB activity,but the results are not statistically significant(P>0.05).4.Histopathological results showed that there was no pathological change in Sham;the myocardial fiber space in Model was enlarged,a large number of red blood cells exuded,and inflammatory cells(mostly neutrophils)infiltrated;800mg/kg significantly improved myocardial tissue damage in rats,with a small amount of infiltration of inflammatory cells between myocardial cells.The pathological morphology of Xinyue capsules 800 mg/kg group was better than that of Xinyue capsules 400 mg/kg group.Xinyue capsules were close in the 800 mg/kg group.5.Compared with Sham,the peripheral blood leukocytes of Model rats increased significantly(P<0.01);compared with Model,Xinyue capsules 400,800 mg/kg and positive drugs could significantly reduce the peripheral blood of rats Number of white blood cells(P<0.01).6.Compared with Sham,the MPO activity in the myocardial tissue of Model was significantly increased(P<0.01).Compared with Model,Xinyue capsules 400,800 mg/kg can significantly reduce the activity of MPO in myocardial tissue(P<0.01),and the positive drug can also reduce the activity of MPO in myocardial ischemiareperfusion rat myocardial tissue(P<0.01).7.There was almost no positive expression of ICAM-1 protein in the myocardial tissue of Sham.Compared with Sham,ICAM-1 positive staining in the myocardial tissue of Model increased significantly,and the IOD/Area ratio increased(P<0.01),And they are all expressed on the inner wall of blood vessels;the expression levels of ICAM-1 in myocardial tissue of rats in the Xinyue capsules 400 and 800 mg/kg groups were significantly reduced compared with Model(P<0.01).CD62 L protein is mainly expressed on the surface of lymphocytes.There was almost no positive expression of CD62 L protein in the myocardial tissue of Sham.Compared with Sham,CD62 L positive staining in the myocardial tissue of Model significantly increased,and the IOD/Area ratio increased(P<0.01),and most of the expressions on the inner of blood vessels;the expression level of CD62 L in myocardial tissues of rats in Xinyue capsules 400 and 800 mg/kg group was significantly reduced compared with Model(P<0.01).8.Compared with Sham,the levels of IL-1?,IL-18,IFN-? and TNF-? in myocardial tissue of Model were significantly increased(P<0.01);In comparison,Xinyue capsules 400 and 800 mg/kg significantly reduced the levels of IL-1? and IFN-? in myocardial tissue of rats(P<0.01),Xinyue capsules 800 mg/kg significantly reduced the IL-18 content in rats(P<0.01),positive drugs can also reduce the content of IL-1?,IFN-? and TNF-? in myocardial tissue of myocardial ischemia-reperfusion no-reflow rats(P<0.01),Xinyue capsules 400,800 mg/kg can reduce TNF-? content,but the results have no statistical significance(P>0.05).9.Compared with Sham,the expression of NLRP3 inflammasome-associated proteins in the myocardial tissue of Model was significantly increased(P<0.01).Compared with Model,Xinyue capsules 400,800 mg/kg can make the myocardium tissue protein expression significantly reduced in a dose-dependent manner(P<0.05 or P<0.01).Positive drugs also reduced the expression of NLRP3 inflammasomeassociated proteins in rat myocardial tissues with significant differences(P<0.01).Conclusion:The results show that Xinyue capsules can significantly reduce the area of myocardial ischemia,infarction and no-reflow in myocardial ischemia-reperfusion nonreflow model rats,significantly improve heart function,reduce myocardial tri-enzyme infiltration and improve the pathological changes of myocardial tissue injury.The mechanism may be reduced the inflammatory response by inhibiting the activation of NLRP3 inflammasome-associated signaling pathways,thereby reducing the accumulation and adhesion of white blood cells in myocardial microvessels,and ultimately reducing no-reflow phenomenon during perfusion. |
PDF Full Text Request