| ObjectiveThe study explored the myocardial protective effect and mechanism of Mongolian medicine zadi-5 on ischemia-reperfusion injury model rats.In order to reveal the myocardial ischemia-reperfusion injury with Mongolian medicine theory,the myocardial protective effect of zadi-5,which is commonly used in clinical coronary heart disease and angina pectoris,is verified by modern scientific experimental methods,so as to provide references for clinical and scientific research.Methods1.Grouping and Administration:84 male SD rats weighing 200 ± 20g were randomly divided into 7 groups according to their body weight:blank group,model group,sham operation group,Chinese medicine control group,Mongolian medicine high-dose group,Mongolian medicine middle dose group and Mongolian medicine low-dose group,with 12 rats in each group.The control group of traditional Chinese medicine was given compound Danshen dripping pill 0.5g/kg diluted in distilled water,Mongolian medicine high,medium and low dose groups were given zadi-5,1.6g/kg,0.8g/kg and 0.4g/kg diluted in distilled water by gavage.The blank group,model group and sham operation group were given the same volume of distilled water 1.0ml/100g,once a day for 14 days.2.Model making and screening:Twelve hours after the last administration,the rat model of Miri was established by ligation of the left anterior descending coronary artery.In the sham operation group,only thread was inserted without ligation.In the other groups,the left anterior descending coronary artery was ligated for 30 minutes and then reperfused for 90 minutes.3.Sampling and testing:ECG was recorded to evaluate whether the model was established or not;blood samples were taken after successful modeling,and the contents of AST,LDH,1dh-1 in serum and CK,MDA,SOD and Ca2+ in myocardial tissue were measured by ELISA.TTC staining was used to detect myocardial infarction and calculate infarct size;TUNEL method was used to detect myocardial apoptosis and calculate apoptosis rate;he staining was used to observe the morphological changes of myocardial tissue;immunohistochemistry was used to detect the expression of Bcl-2,Bax and caspase-3;QRT PCR was used to detect the expression of miR-30a in myocardial tissue.4.Statistical treatment:SPSS 22.0 statistical software was used for statistical x±S).One way ANOVA was used,and tamhane test was used for uneven variance.The test level was p=0.05,and P<0.05 was statistically significant.Results1.The results of electrocardiography and HE stainingST segment was elevated in lead ? of rat limb during ischemia by ligation of anterior descending branch of left coronary artery,while ST segment gradually returned to normal during reperfusion,suggesting that myocardial ischemia-reperfusion injury model was successfully established.It was observed by HE staining that the myocardial tissue cells in the model group were disordered,cell rupture,necrosis,inflammatory cell infiltration,myocardial fiber rupture,while the traditional Chinese medicine group and Mongolian medicine group reversed the above changes in different degrees.2.Effect of Mongolian medicine zadi-5 on myocardial infarction area in Miri model ratsCompared with the blank control group,the myocardial infarction area of model group was significantly increased(P<0.05),and the myocardial infarction area was significantly larger than that of the sham operation group(P<0.05).Compared with the model group,the myocardial infarction area of rats in the TCM control group and Mongolian medicine high and medium dose groups was significantly reduced(P<0.05),while the myocardial infarction area of rats in low-dose Mongolian medicine group showed a decreasing trend,but there was no statistical significance(P>0.05).3.The effect of Mongolian medicine zadi-5 on ast,LDH,ldh-1 in serum of Miri model ratsCompared with the blank control group,the contents of AST,LDH and ldh-1 in serum of model group were significantly increased(P<0.05),and the contents of AST,LDH and ldh-1 in serum of model group were significantly higher than those in sham operation group(P<0.05).Compared with the model group,the contents of AST,LDH and ldh-1 in serum of rats in TCM control group and Mongolian medicine high and medium dose groups were significantly decreased,with statistical significance(P<0.05).The contents of AST,LDH and ldh-1 in serum of rats in low-dose Mongolian medicine group showed a downward trend,but there was no statistical significance(P>0.05).4.Effect of Mongolian medicine zadi-5 on CK,MDA,SOD,Ca2+ content in myocardial tissue of Miri model ratsCompared with the blank control group,the content of CK,MDA and Ca2+ in myocardial tissue of model group was significantly increased,and the content of SOD was significantly decreased(P<0.05).The content of CK,MDA and Ca2+in myocardial tissue of model group was significantly higher than that of sham operation group,and the content of SOD was significantly lower than that of sham operation group(P<0.05).Compared with the model group,the contents of CK and Ca2+in the control group and the high and middle dose groups of Mongolian medicine were significantly decreased,and the content of SOD was significantly increased(P<0.05),and the content of MDA in the high-dose group was significantly decreased(P<0.05).The content of MDA in middle dose group and CK,MDA and Ca2+in low dose group decreased,while SOD content increased,but there was no statistical significance(P>0.05).5.The effect of Mongolian medicine zadi-5 on the expression of Bcl-2,Bax,caspase-3 in myocardial tissue of Miri model ratsCompared with the blank control group,the expression of Bax and caspase-3 protein in myocardial tissue of model group was significantly increased,and the expression of Bcl-2 protein was significantly decreased(P<0.05).The expression of Bax and caspase-3 protein was significantly higher than that of sham operation group,and the expression of Bcl-2 protein was significantly lower than that of sham operation group(P<0.05).Compared with the model group,the expression of Bcl-2 protein was significantly increased,Bax and caspase-3 protein expression were significantly decreased in the Chinese medicine control group and Mongolian medicine high and medium dose groups(P<0.05),and the expression of Bcl-2 protein in myocardial tissue of low-dose Mongolian medicine group was significantly increased(P<0.05).The expression of Bax and caspase-3 protein in myocardial tissue of low-dose Mongolian medicine group decreased,but there was no statistical significance(P>0.05).6.The effect of Mongolian medicine zadi-5 on the apoptosis rate of Miri model ratsCompared with the blank control group,the apoptosis rate of myocardial cells in the model group was significantly higher than that in the sham operation group(P<0.05).Compared with the model group,the apoptosis rate of myocardial cells in the Chinese medicine control group and Mongolian medicine high and medium dose groups was significantly decreased(P<0.05).The apoptosis rate of myocardial cells in low-dose Mongolian medicine group decreased,but there was no statistical significance(P>0.05).7.The effect of Mongolian medicine zadi-5 on the expression of miR-30a in Miri model ratsCompared with the blank group,the expression of miR-30a in the model group was significantly decreased(P<0.05),and the expression of miR-30a in the model group was significantly lower than that in the sham operation group(P<0.05).Compared with the model group,the expression of miR-30a in TCM control group and Mongolian medicine high and medium dose groups was significantly increased(P<0.05).The expression of miR-30a in the low-dose group of Mongolian medicine increased,but there was no statistical significance(P>0.05).Conclusion1.Mongolian medicine zadi-5 can decrease the contents of AST and LDH in serum and CK and Ca2+in myocardial tissue on Miri rats.Also it can reduce the degree of myocardial cell membrane damage,the ion permeability and resist calcium overload.Mongolian medicine zadi-5 plays a protective role in myocardium.2.Mongolian medicine zadi-5 can decrease the content of MDA and increase the activity of SOD in myocardial tissue on Miri rats.It can effectively improve the activity of antioxidant enzymes and the ability of scavenging oxygen free radicals.Also it can inhibit the production of oxygen free radicals.Mongolian medicine zadi-5 plays a protective role by inhibiting lipid peroxidation of myocardial cell membrane.3.Mongolian medicine zadi-5 can increase the expression of Bcl-2 protein,decrease the expression of Bax protein,reduce the number of apoptotic positive cellsand the apoptosis rate.It can effectively enhance the inhibition of apoptosis factors,reduce the pro apoptotic factors.Mongolian medicine zadi-5 plays a protective role by inhibiting apoptosis.4.Mongolian medicine zadi-5 can increase the expression of miR-30a and the level of Bcl-2 on Miri rats,which can protect myocardium by reduce the gene regulation andcell apoptosis rate.5.The Mongolian medicine zadi-5 can regulate the blood and blood phase,improve the blood and blood circulation,enhance the cardiac function by regulating the complement and voxel,and protect the myocardium. |
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