| Objective:In this study,the rats were effectively intervened with the Mongolian medicine Zhu Ri Hen dripping pills,the objective is to clarify the effect of Zhu Ri Hen Dripping Pills in myocardial ischemia/reperfusion injury and its molecular mechanism,thus,to provide an objective basis for Zhu Ri Hen Dripping Pills in the treatment of Xin Ci Tong pain in Mongolian medicine.Methods:Male SD rats were randomly divided into the sham operation group(Sham),model group(I/R),positive group(DS),Zhu Ri Hen Dripping Pills low(ZRH-L),medium(ZRH-M),high(ZRH-H)dose groups;Rats in each group were given intragastrically for 15 days continuously,once a day,rats in the sham group and I/R group were given normal saline(10ml·kg-1)respectively,rats in the DS group were given Composite Salvia Dropping Pill(170 mg·kg-1),and rats in the ZRH-L,ZRH-M,and ZRH-H groups were given Zhu Ri Hen Dripping Pills(300,900,1500 mg·Kg-1)respectively.Rats in each group were administered once one hour before modeling.The left anterior descending coronary artery in rats was ligated for 30min for ischemia and reperfusion for 2h.The model myocardial ischemia/reperfusion in rats was constructed.Blood was collected from the abdominal aorta,the heart was removed,and the specimen was kept.Image analysis software(mage-Pro Plus)was used to calculate the area of myocardial infarction,enzyme-linked immunosorbent assay(ELISA)method was used to detect the content of serum creatine kinase isoenzyme MB(CK-MB),cardiac troponin T(CTnT)and the content of myocardial malondialdehyde(MDA),the activity of superoxide dismutase(SOD).HE staining method was used to observe the pathological morphological changes of myocardial tissues and transmission electron microscopy was used to observe the ultrastructural changes of myocardial mitochondria,and the protective effect of Zhu Ri Hen Dripping Pills was evaluated.The TUNEL method was used to detect and calculate the apoptosis index of cardiomyocytes and observe the effect of Zhu Ri Hen Dripping Pills on cell apoptosis;Western Blot method was used to detect the expression levels of Cyto-C,c-caspase-9,c-caspase-3,Bax,Bcl-2,P-Akt protein respectively and RT-PCR mwthod was used to detect the expression levels of Bax mRNA and Bcl-2 mRNA.The mechanism of Zhu Ri Hen Dripping Pills was clarified.Results:1 The effect of Mongolian medicine Zhu Ri Hen Dripping Pills on the area of myocardial infarction after MIRI:the results showed that compared with the Sham group,the area of myocardial infarction in the I/R group was significantly increased;Compared with the I/R group,the area of myocardial infarction in the ZRH-M group,the ZRH-H group and the DS group was decreased significantly,and the difference was statistically significant(P<0.001);There was no statistical significance in the area of myocardial infarction between the ZRH-M group and the DS group(P>0.05).2 The effect of Mongolian medicine Zhu Ri Hen Dripping Pills on the content of serum CK-MB and CTnT after MIRI:the results showed that compared with the Sham group,the content of CK-MB and CTnT in the I/R group was increased significantly;Compared with the I/R group,the content of CK-MB and CTnT in the ZRH-M group,ZRH-H group and DS group was decreased significantly,and the difference was statistically significant(P<0.001);There was no statistical significance in the content of CK-MB and CTnT between the ZRH-M group and the DS group(P>0.05).3 The effect of Mongolian medicine Zhu Ri Hen Dripping Pills on the pathological morphology of myocardial tissues and myocardial mitochondrial ultrastructure of rats after MIRI:The observation results under the microscope showed that the morphology of the myocardial cells was irregular,the arrangement was disordered,the local muscle fiber stripes disappeared,and the edema of the myocardial cells was visible,and there was focal degeneration and necrosis,neutrophil cells were infiltrated in the I/R group.In each group of Zhu Ri Hen Dripping Pills and the DS group,myocardial cell edema was reduced,and the number of neutrophil infiltration was reduced.The observation results under transmission electron microscope showed that in the I/R group,the arrangement of myofilaments was loose and disordered;Most of mitochondria were swollen,the ridge gap increased,and the ridge fracture was accompanied.In each group of Zhu Ri Hen Dripping Pills and the DS group,there was no obvious loosening of myofilament,the size and shape of mitochondria were more normal,mild to moderate swelling,the ridge arrangement was basically ordered,and the structure was still clear and complete.4 The effect of Mongolian medicine Zhu Ri Hen Dripping Pills on the content of myocardial MDA and the activity of SOD after MIRI:the results showed that compared with the Sham group,the MDA value in the I/R group was significantly increased,and the SOD value was significantly reduced;Moreover,Compared with the I/R group,the MDA value in the ZRH-M group,ZRH-H group and DS group was decreased significantly,and the SOD value was increased significantly,the difference was statistically significant(P<0.001);There was no statistical significance in the MDA and SOD values between the ZRH-M group and the DS group(P>0.05).5 The effect of Mongolian medicine Zhu Ri Hen Dripping Pills on the apoptosis index of cardiomyocytes after MIRI:the results showed that compared with the Sham group,the apoptosis index of cardiomyocytes in the I/R group was:increased significantly;Compared with the I/R group,the apoptosis index of cardiomyocytes in the group ZRH-M,ZRH-H group,and DS group was decreased significantly,the difference was statistically significant(P<0.001);There was no statistical significance in the apoptosis index of cardiomyocytes between the ZRH-M group and the DS group(P>0.05).6 The effect of Mongolian medicine Zhu Ri Hen Dripping Pills on the expression of cyto-C Caspase-9 caspase-3 protein in myocardial after MIRI:the results showed that compared with the Sham group,the expression of Cyto一C,c-caspase-9,c-caspase-3 protein in the I/R group was increased significantly;Compared with the I/R group,the expression of Cyto-C,c-caspase-9,c-caspase-3 protein in the ZRH-M group,ZRH-H group,and DS group was reduced significantly,the difference was statistically significant(P<0.001);Compared with the DS group,the expression of Cyto-C protein in the ZRH-M group and ZRH-H group had no statistical significance(P>0.05);Compared with the DS group,the expression of caspase-9 and c-caspase-3 protein in the ZRH-M group had no statistical significance(P>0.05).7 The effect of Mongolian medicine Zhu Ri Hen Dripping Pills on bcl-2 Bax protein and mRNA expression in myocardial after MIRI:the results showed that Compared with the Sham group,the expression of Bcl-2 protein and mRNA in the I/R group decreased significantly.However,the expression of Bax protein and mRNA was increased significantly;Compared with the I/R group,the expression of Bcl-2 protein and mRNA in the ZRH-M group,ZRH-H group,and DS group was increased significantly,however,the expression of Bax protein and mRNA decreased significantly,the difference was statistically significant(P<0.001);The expression of Bcl-2 protein and mRNA in the ZRH-M group and ZRH-H group had no statistically significant compared with the DS group(P>0.05);Compared with the DS group,the difference of Bax protein expression in the ZRH-M group and ZRH-H group had statistical significance(P<0.05);Compared with the DS group,the expression of Bax protein and mRNA in the ZRH-M group had no statistical significance(P>0.05).8 The effect of Mongolian medicine Zhu Ri Hen Dripping Pills on P-Akt protein expression in myocardial after MIRI-the results showed that Compared with the Sham group,the expression of P-Akt protein in the I/R group was increased significantly;Compared with the I/R group,the expression of P-Akt protein in the ZRH-M group and the DS group was decreased significantly.The difference had statistical significance(P<0.001).Conclusion:1 Mongolian medicine Zhu Ri Hen Dripping Pills can significantly improve myocardial ischemia/reperfusion injury in rats.2 In the process of myocardial ischemia/reperfusion injury,Mongolian medicine Zhu Ri Hen Dripping Pills play a protective role in myocardium by inhibiting the mitochondrial apoptosis pathway.3 PI3K/AKT signaling pathway may be involved in the protective effect of Mongolian medicine Zhu Ri Hen Dripping Pills on myocardial ischemia/reperfusion injury in rats.4 Mongolian medicine Zhu Ri Hen Dropping Pills has the effect of treating the Xin Ci Tongoing of Mongolian medicine. |
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