| Purpose:To investigate the protective effect of yimai granules on hyperlipidemia with myocardial ischemia reperfusion injury(MIRI)in rats.Taking inflammation,oxidative stress and apoptosis mediated by Nrf2/HO-1 pathway as the entry point,this study aims to reveal the protective effect and molecular mechanism of yimai granules on high-fat MIRI.Material and method:1.Protective effect of yimai granule on myocardial injury in rats with high fat and MIRI(1)100 rats were randomly divided into 5 groups,including sham operation group,model group,high,medium and low dose yimai granules group,with 20 rats in each group.Animals in the sham operation group were raised routinely,and the rest groups were fed with high-fat diet for 2 weeks,other conditions being the same.Rats in the sham operation group and model group were given normal saline 10ml/kg daily,while those in the low-dose,medium-dose and high-dose groups were given Chinese herbal decoction 10 ml/kg daily,and the number of times of gavage was 2 times a day.And the rats were given a continuous administration for 1 week before ischemia-reperfusion.Each rat myocardial ischemia reperfusion after 2 hours,the anterior descending coronary artery ligation of left in situ,after waiting for dye distribution to the whole heart tissue anterior descending(perfusion area except),rapid heart,saline flushing,frozen in-20 ?,after myocardial infarction area is determined.At the same time,the rat abdominal aorta was taken for blood,and the serum was separated for later use.Myocardial infarction area is determined based on the rest of the heart tissue cryopreserved-80 ? cryogenic refrigerator.(2)Cholesterol level(TG,TC,HDL-C and LDL-C)was detected by automatic biochemical analyzer.(3)Myocardial enzyme profiles(CK,CK-MB,HBDH and LDH)were detected by ultraviolet spectrophotometer.(4)TTC method was used to detect the myocardial infarction area of rats in each group.(5)HE staining was used to detect the morphological changes of the heart tissue in each group.(6)The changes of ST segment and T wave of ECG in each group were analyzed.2.Effect of yimai granules on myocardial inflammatory factors in rats with high lipid and MIRI(1)Modeling and grouping is same as above.(2)Serum TNF-?,IL-6,IL-1 were detected by ELISA.(3)mRNA expression of TNF-?,IL-6 and IL-1? in rat myocardial tissue was detected by RT-qPCR.(4)The expression of NF-?B protein was determined by Western blot.3.Yimai granules affect oxidative stress and apoptosis of high-fat MIRI rats through Nrf2/HO-1 pathway(1)modeling and grouping is same as above.(2)Fluorescence probe method was used to determine the ROS activity in rat cardiac tissue.(3)The levels of MDA,SOD and GSH-px in the serum of rats in each group were determined by ELISA.(4)RT-qPCR method was used to detect the expressions of Nrf2 and HO-1 mRNA in rat cardiac tissue.(5)The expressions of Bax,Bcl-2,Caspase3,Nrf2 and HO-1 in myocardium of rats in each group were determined by Western blot.Results: 1.Serum lipid level test showed that,compared with the model group,the high dose group of yimai granules could significantly reduce the serum TG,TC,LDL-C level and significantly increase the HDL-C level in rats(compared with the model group,P <0.01).2.The detection of serum myocardial enzyme spectrum showed that the serum CK,CK-MB,HBDH and LDH levels of rats in the high-dose group of yimai granules could be significantly reduced(P <0.01 compared with the model group).3.HE staining showed that in the high-dose group of yimai granules,there was only a small amount of necrotic area,myocardial fibers were arranged in a orderly manner,and there was only a small amount of inflammatory cell infiltration in myocardial interstitium with clear structure,which could significantly improve the pathological changes of myocardial tissue MIRI injury.4.Myocardial infarction area detection showed that the high-dose yimai granules group could significantly reduce the myocardial infarction area of rats(compared with the model group,P <0.01).5.The analysis of electrocardiogram results showed that high dose of yimai granules could significantly reduce the elevated ST segment and T wave level of MIRI rats(compared with the model group,P <0.01).6.ELISA results showed that the high-dose group of yimai granules could significantly reduce the expression of TNF-?,IL-6,and IL-1? in the serum of MIRI rats(compared with the model group,P <0.01).7.The RT-qPCR results showed that the high-dose yimai granules group could significantly reduce the expression of TNF-?,IL-6,and IL-1? mRNA in rat myocardial tissue(compared with the model group,P <0.01).8.Western blot results showed that the high dose yimai granules group could significantly reduce the expression level of NF-?B protein in rat myocardial tissue(P <0.01 compared with the model group).9.The ROS test results by fluorescent probe showed that the high dose group of yimai granules could significantly lower the ROS level(P <0.01 compared with the model group).10.ELISA results showed that the high-dose group of yimai granules could significantly increase serum SOD and GSH-px levels,and significantly reduce MDA levels in rats(compared with the model group,P <0.01).11.Western blot results showed that the high dose group of yimai granules significantly increased the level of BCl-2 in rat cardiac tissue,and significantly decreased the levels of Bax and Caspase3(compared with the model group,P <0.01).12.Western blot results showed that the high-dose group of yimai granules significantly increased the expression levels of Nrf2 and HO-1 proteins in rat cardiac tissue(compared with the model group,P <0.01).13.RT-qPCR results showed that the high-dose yimai granules group could significantly improve the expression levels of Nrf2 and HO-1 mRNA in rat cardiac tissue(compared with the model group,P <0.01).Conclusion: 1.Yimai granules can play a role in improving dyslipidemia and alleviating myocardial ischemia reperfusion injury in rats with high lipid and myocardial ischemia reperfusion injury.2.Yimai granules can improve myocardial injury and play a protective role by inhibiting inflammatory reaction,oxidative stress and apoptosis.3.Yimai granules can inhibit oxidative stress and apoptosis to improve myocardial ischemia reperfusion injury by regulating the Nrf2/HO-1 pathway. |
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