| Objective:In this study,the effects and mechanism of Buyang Huanwu decoction on MI/RI in rats were investigated by detecting cardiac function,cell apoptosis and expression of related pathway proteins in myocardial ischemia/reperfusion rats.Materials and methods:1.Fifty male SD rats were randomly divided into Sham operation group(Sham),myocardial ischemia reperfusion group(IR),Buyang Huanwu decoction low-dose group(LBYHW),Buyang Huanwu decoction medium-dose group(MBYHW)and Buyang Huanwu decoction high-dose group(HBYHW),with 10 rats in each group.The LBYHW group,MBYHW group and HBYHW group were given three different doses of Buyang Huanwu decoction by gavage every day,and the Sham group and IR group were given normal saline by gavage every day.The rats in the five groups were given 2 ml of each gavage and were pretreated by gavage for 7 days.The model of myocardial ischemia-reperfusion in vivo was established by ligation of the anterior descending coronary artery(LAD).Ecg monitoring was carried out on the rats during the modeling with limb lead electrocardiogram to determine whether the model of ischemia reperfusion was successful,and the changes of electrocardiogram during the modeling were observed.2.Cardiac function was evaluated by color Doppler ultrasound 3 h after reperfusion.After the examination,venous blood and heart were taken for relevant detection,that is,HE staining was performed to compare the changes of myocardial cell structure in each group,serum lactate dehydrogenase(LDH)concentration was detected by ELISA,and apoptosis was detected by TUNEL staining.Through the above detection methods,the differences in cardiac function and myocardial cells after MI/RI were observed in each group.3.The hearts of rats were taken 3 h after reperfusion,and the expressions of SIRT1/Fox O1 signaling pathway proteins and necrotic apoptosation-related proteins RIP1,RIP3 and MLKL were detected by Western blot.Immunofluorescence assay was used to detect the expression level of SIRT1 protein,so as to explore the effect of Buyang Huanwu Decoction on SIRT1/Fox O1 signaling pathway and its possible mechanism.Results:1.The results of echocardiography showed that compared with Sham group,the cardiac function indexes of LVEF and LVFS in IR group were significantly decreased,and the difference was statistically significant(P < 0.05).Compared with IR group,LBYHW group,MBYHW group and HBYHW group had improved LVEF and LVFS,and the difference was statistically significant(P < 0.05).These results indicate that Buyang Huanwu decoction pretreatment can alleviate the MI/RI induced cardiac function decline and improve the cardiac function of rats.2.HE staining and TUNEL staining were used to detect myocardial cell injury and apoptosis.The results showed that,compared with the Sham group,the myocardial tissue injury and the number of myocardial cell apoptosis were significantly increased in IR group.Compared with IR group,the myocardial tissue damage of LBYHW group,MBYHW group and HBYHW group was alleviated,and the number of myocardial cell apoptosis was decreased to varying degrees.Among the four groups of model group,the MBYHW group had the closest cell arrangement,the least inflammatory infiltration phenomenon,and the lowest number of myocardial cell apoptosis(P < 0.05).3.ELISA showed that compared with Sham group,serum LDH concentration in IR group was significantly increased(P < 0.05).Compared with IR group,the serum LDH concentration of LBYHW group,MBYHW group and HBYHW group was decreased,and the serum LDH level of MBYHW group and IR group was significantly different(P < 0.05).4.Western blot analysis showed that compared with Sham group,the protein expressions of RIP1,RIP3,MLKL and Ac-Fox O1 in myocardial tissue of IR group were significantly increased(P < 0.05),while the protein expressions of SIRT1 were significantly decreased(P < 0.05).Compared with IR group,the expression levels of RIP1,RIP3,MLKL and Ac-Fox O1 in LBYHW group,MBYHW group and HBYHW group were decreased,while the expression levels of SIRT1 protein were increased,especially in MBYHW group(P < 0.05).5.Immunofluorescence staining results showed that SIRT1 protein expression in myocardial tissue of rats in IR group was significantly decreased compared with Sham group(P < 0.05).Compared with IR group,SIRT1 protein expression levels of LBYHW group,MBYHW group and HBYHW group were increased,and the increase effect of MBYHW group was the most significant(P < 0.05).Conclusion:1.Buyang Huanwu Decoction pretreatment can improve myocardial function indexes after MI/RI and reduce the damage of MI/RI to myocardial cells and structures.2.Buyang Huanwu Decoction pretreatment can reduce the expression of RIP1,RIP3,MLKL necrosis protein after MI/RI,and inhibit the necrotic apoptosis of rat cells.3.Buyang Huanwu Decoction may play a role in preventing MI/RI by regulating SIRT1/Fox O1 signaling pathway. |
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