| Objective: To investigate the effect of Miaoling natto capsule pretreatment on myocardial ischemia-reperfusion injury in rats and its possible mechanism.Determining the contents of the main active ingredients in the natto capsule of Miaoling.Methods: 1.90 healthy male SD rats were randomly divided into the sham group,the model group,positive control group,Miaoling natto capsules low,medium and high does groups.Using ligation the left anterior descending coronary artery of rat heart,the method of myocardial ischemia-reperfusion model was set up.(1)The standard ECG leads II segment of ST and T waves values for indicators was observed in rat with myocardial ischemia-reperfusion electrocardiogram effect.(2)To observe and messure myocardial infarction area with TTC staining,To observation mycocardial cell structure and inflammatory cells infiltration with HE dyeing.(3)Determination of aspartate aminotransferase in serum the levels of acetic transaminase(AST),lactate dehydrogenase(LDH),creatine kinase isoenzyme(CK-MB),cardiac troponin-I(cTn-I).(4)Detect myocardial ischemiareperfusion serum content of superoxide dismutase(SOD),malondialdehyde(MDA),and tumor necrosis factor-?(TNF-?).2.The fibrin plate method was used to determine the plasmin activity of Miao ling natto capsules.The content of total flavonoids in soybean was determined by UV method.The compounds containing daidzin,genistin,daidzein,and genistein were determined by HPLC.The content of soybean polysaccharide was determined by phenol-sulfuric acid colorimetry,and the content of soybean saponin was determined by vanillin-perchloric acid colorimetry.Results: 1.(1)Compared with the sham group,the heart rate of the model group increased significantly during the ischemic period(P<0.05).There was no significant difference in heart rate after the ligature was released.Compared with the model group,there was no statistically significant difference in the heart rate between the high,middle,and low doses of Miaoling natto capsules.(2)Compared with model group,the Miaoling natto capsules groups against myocardial ischemia-reperfusion at the stage of ischemia phases of ST segment and T waves elevation had significant differences(P<0.05),there was no significant difference in reperfusion time(P>0.05).(3)Compared with sham group,myocardial infarct size of model group at the end of reperfusion was significantly increased(P<0.001),the Miaoling natto capsules groups at the end of reperfusion,infarct size was significantly reduced(P<0.001).(4)Compared with model group,visible muscle fiber fracture,disorder,myocardial cell gap widened,shrink phenomenon,which has a bleeding,nucleus pycnosis,burst;Miaoling natto capsulesc low,medium and high does groups visible muscle fiber arrangement is neat,dense,not seen obvious fracture,myocardial cells arranged relative rules,no obvious hypertrophy,edema in drug groups.(5)Miaoling natto capsule groups can significantly reduce the LDH,CK-MB activity(P <0.05),for the AST activity,the high dose group can significantly reduce its activity(P <0.05),For the cTnI content,there was no significant difference in the low dose group(P>0.05).(6)Compared with the model group,Miaoling natto capsule groups can significantly increase the activity of SOD and decrease the content of MDA(P<0.05).For the content of TNF-?,the high dose group can significantly reduce the content of TNF-?(P<0.05).2.The determination of the main active substances: The fibrin plate method was used to measure the activity of thrombolytic kinase in the Miaoling natto capsule: 3406.01 IU/g;the UV was used to determine the isoflavone content: 4.80 mg/g;the HPLC method was applied to determine the four kinds of soybean isoflavones,daidzin 0.644 mg/g,genistein 0.733 mg/g,daidzein 0.213 mg/g,genistein 0.095 mg/g;sulfuricacid-phenol method for the determination of soybean polysaccharide: 6.38mg/g;UV method of soybean saponin content: 17.18mg/g.Conclusion: 1.Through the experiment this study finds that Miaoling natto capsule can make the rats of myocardial ischemia-reperfusion injury with no significant changed in heart rate,ST segment and T wave amplitude decreased;the infarct size reduced;it can improve the cell morphology,reduce the myocardial interstitial edema,and improve fibrils arranged in a disordered condition;it can significantly reduce the levels of serum AST,LDH,CK-MB and cTnI content.The experimental results suggest that the drug has protective effect on myocardial ischemia-reperfusion injury.2.The study found that Miaoling natto capsule increased the activity of SOD and reduced the content of MDA and TNF-?.Its mechanism of protective effect may be by inhibiting of the production of free radicals,reducing lipid peroxide,maintaining the homeostasis of myocardial cell oxidation and antioxidants,and reducing inflammation factor synthesis or releasing.3.In this experiment,the main active substances in Miaoling nattocapsule were determined.Simultaneously,the content of the four kinds of soybean isoflavones was determined by HPLC,which provided the material basis for the protective effect of Miaoling Natto Capsule on myocardial ischemia-reperfusion injury. |
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