| Objective:To study the effect of Yindan Xinnaotong Soft Capsule preconditioning on myocardial ischemia reperfusion injury in SD rats by establishing myocardial ischemia reperfusion injury model;To study whether Yindanxinnaotong soft capsule could alleviate myocardial ischemia reperfusion injury in rats by regulating Bcl-2/Bax/Caspase-3 signaling pathway to inhibit apoptosis.Methods:SD rats were randomly divided into 14 rats in each group,After continuous gavage for 2 weeks,the myocardial ischemia-reperfusion injury model of SD rats were established by ligating the left anterior descending branch(LAD)for 15 minutes and loosening the reperfusion for 2 h.The specific groups were as follows:(1)Sham operation group: 1 m L normal saline was continuously perfused for 2 weeks,and then the chest was opened without ligation;(2)Model group: 1 m L normal saline was continuously perfused for 2 weeks,then the chest was opened,LAD was ligated for15 minutes,and the rats were released for reperfusion for 2 h;(3)Yindan low dose group: Yindan Xinnaotong Soft Capsule was dissolved in normal saline.After continuous gavage at the dose of 200 mg/kg for 2 weeks,LAD was ligated for 15 minutes,and then released for 2 h of reperfusion;(4)Yindan middle dose group:Yindan Xinnaotong Soft Capsule was dissolved in normal saline,gavaged continuously at the dose of 400 mg/ kg for 2 weeks,LAD ischemia of rats was ligated by thoracotomy for 15 minutes,and then released for reperfusion for 2 h;(5)Yindan high dose group: Yindan Xinnaotong Soft Capsule was dissolved in normal saline,after continuous gavage at the dose of 800 mg/kg for 2 weeks,LAD was ligated for15 minutes,and then released for reperfusion for 2 h.The changes of electrocardiogram(ECG)of rats in each group were detected in the whole process of modeling.After successfully constructing the myocardial ischemia reperfusion injury model of SD rats,HE staining was used to observe the morphology of cardiomyocytes;TTC staining was used to evaluate the extent of myocardial infarction;Left Ven-tricular end-diastolic Inner Dimension(LVIDd),Left Ventricular end-systolic Inner Diameter(LVIDs),Left Ventricular Ejection Fraction(LVEF),Left Ventricular Fractional Shortening(LVFS)and Stroke Volume(SV)were measured by echocardiography in rats of each group;The positive expression levels of Bcl-2 and Bax were detected by immunohistochemistry;The expression levels of Bcl-2,Bax and Cleaved caspase-3 were detected by Western blotting.Results:The results of HE showed that the myocardial cells in the Sham operation group were orderly arranged with clear boundaries,the gap of myocardial cells in the Model group was widened,the myocardial cells were irregularly arranged,myocardial fibers were broken and inflammatory cells infiltrated,and the degree of myocardial cell injury in the Yindan groups with different doses was reduced,among them,the degree of cell injury in the Yindan high dose group was the least,followed by the Yindan medium dose group,and finally the Yindan low dose group.TTC results showed that there was no infarct in the Sham operation group;Compared with the Sham operation group,the myocardial infarction area in the Model group increased significantly(P<0.05);Compared with the Model group,the myocardial infarction area of different doses of Yindan group decreased(P<0.05),with the increase of Yindan dose,the myocardial infarction area became smaller.The results of cardiac ultrasound in rats showed that compared with the Sham operation group,LVEF,LVFS and SV in the Model group were significantly lower(P<0.05);Compared with the Model group,LVEF,LVEF and SV of different doses of Yindan group increased(P<0.05),with the increase of Yindan dose,LVEF and LVFS increased,while the SV result was that the value of Yindan medium dose group was the highest,followed by Yindan high dose group,and finally Yindan low dose group;Compared with the Sham operation group,LVIDs in the Model group increased(P<0.05);Compared with the Model group,LVIDs in different doses of Yindan group decreased,the higher the dose,the lower the LVIDs;The LVIDd results of Sham operation group,Model group,Yindan low dose group,Yindan medium dose group and Yindan high dose group had little difference(P>0.05),The measurement results of cardiac ultrasound in rats showed that different doses of Yindan Xinnaotong could significantly improve the cardiac function of MIRI rats,The higher the dose of Yindan,the more obvious the improvement of cardiac function.Immunohistochemical results showed that compared with Sham operation,Bcl-2 decreased significantly(P<0.05),Bax increased significantly(P<0.05),and Bcl-2/ Bax ratio decreased in the Model group;Compared with the Model group,different doses of Yindan Xinnaotong Capsule could significantly reduce the expression of Bax(P<0.05),increase the expression of Bcl-2(P<0.05)and the ratio of Bcl-2/Bax.Western blot showed that compared with the Sham operation group,the expression of Bcl-2 protein in the Model group decreased,but the difference was not statistically significant(P>0.05),while the expression of Bax and Cleaved caspase-3 increased significantly(P<0.05),and the ratio of Bcl-2/Bax decreased(P<0.05);Compared with the Model group,different doses of Yindan Xinnaotong Capsule could reduce the expression of Bax and Cleaved Caspase-3,increase the expression of Bcl-2 and the ratio of Bcl-2/Bax(P<0.05).Conclusions:Yindan Xinnaotong Soft Capsul pretreatment can inhibit apoptosis and reduce MIRI in rats by regulating Bcl-2/Bax/caspase-3 signal pathway. |
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