| Objective:By using high-fat diet combined with injection of isoproterenol+chronic unpredictable stress method to establish the model,simulate the high-fat state of myocardial ischemia combined with depression rat model,using modified Chaihu Shugan powder and its disassembled formula for drug treatment.The rat model of double heart disease was evaluated by detecting blood lipid,he staining and behavior.ACE2 ang(1-7)-Masr axis was selected as the breakthrough point.The contents of angiotensin Ⅱ(AngⅡ),angiotensin(1-7)[ang(1-7)]and angiotensin converting enzyme 2(ACE2)in rat serum were detected by ELISA,and the gene and protein expressions of ACE2 and Masr in hippocampus and heart were detected by RT-PCR and Western blot,Objective to provide a new target basis for modified Chaihu Shugan powder in the prevention and treatment of myocardial ischemia combined with depression,and provide a new idea for the clinical prevention and treatment of the disease with traditional Chinese medicine.Methods:Experiment 1:30 male SD rats were randomly divided into three groups:normal group,model group and drug group(modified Chaihu Shugan powder group),with 10 rats in each group.The rats in the normal group were fed with common food and were not treated;The rats in model group and drug group were fed with high-fat diet(3%cholesterol,10%lard,0.2%propylthiouracil,0.5%sodium cholate,5%sugar and 81.3%basic diet)on the first day,At the same time,the animals were exposed to various kinds of unpredictable stress,including behavior restriction,heat shock,moist bedding,45℃ cage tilt,cage shaking,day and night reversal,ice water swimming,noise 85dB and so on.At the 6th week,ISO solution 3mg/kg was injected intraperitoneally daily for 21 days.The model was established for 8 weeks.The drug group was treated with modified Chaihu Shugan powder from the first day to the end of modeling,and the normal group and model group were treated with the same amount of normal saline.The depression was evaluated by behavioral observation;Four items of blood lipids were detected and the blood lipids were observed;He staining was used to observe the pathological changes of heart tissue.In order to evaluate the success of the model.Experiment 2:108 male SD rats were randomly divided into 9 groups:normal group,model group,modified Chaihu Shugan powder group,Quyu Huatan Decoction group,Shugan Xingqi Decoction group,Jianpi Yangxue Decoction group,fluoxetine group,trimetazidine group and simvastatin group,with 12 rats in each group.The modeling method of model group and each drug group was the same as the experiment.From the first day to the end of modeling,each drug group was given corresponding drug treatment.After modeling,the levels of ACE2,ang(1-7),AngⅡ in serum were detected by ELISA,and the gene and protein expressions of ACE2 and Masr in heart and hippocampus were detected by Real-time PCR and Western blot.As a breakthrough point to explore the effect of modified Chaihu Shugan powder and its disassembled formula on ACE2 ang(1-7)-Masr axis in rats with myocardial ischemia and depression in hyperlipidemic state.Result:Experiment 1:1.General condition:the rats in the normal group have bright eyes,sensitive reactions,powerful activities,full muscles,smooth body hair,and normal urine and feces.After modeling,the model group was manic,hissing,struggling to break away,moving around,losing weight significantly,loss of appetite,slowly showing mental depression,fatigue,soft stools,rough hair color,dark purple tongue.Compared with the model group,the rats in the drug group were significantly better in state,active,flexible in action,moistening in coat color,forming in stool,increasing in appetite and increasing in weight.2.Behavior:compared with the normal group,the movement time,distancein the central area and standing times of the open field test of the model group were significantly reduced(P<0.01),the immobility time of the forced swimming test was significantly increased(P<0.01).Compared with the model group,the movement time,distance in the central area and standing times of the open field test in the modified Chaihu Shugan powder group were significantly higher than those in the model group(P<0.01),the immobility time of the forced swimming test was significantly lower(P<0.01)3.Hematoxylin eosin staining method:the morphology of heart in each group was observed under microscope,and the morphology of myocardial fibers in normal group was complete and arranged regularly.In the model group,myocardial cells were injured and necrotic,myocardial fibers were disordered,fibers were broken,and inflammatory cells were infiltrated.The morphology of modified Chaihu Shugan powder group was basically complete,and the arrangement of myocardial cells was more regular and orderly than that of model group.4.Four items of blood lipid:compared with the normal group,the levels of TC and LDL-C in the model group increased,while the level of HDL decreased(P<0.01),and there was no significant difference in TG(P>0.05).Compared with the model group,the contents of TC and LDL-C in the modified Chaihu Shugan powder group were lower than those in the model group,while the contents of HDL-C in the modified Chaihu Shugan powder group were higher than those in the model group(P<0.01),and there was no significant difference in TG(P>0.05).Experiment 2:1.Serum AngⅡ,ACE2 and Ang(1-7)levels:modified Chaihu Shugan powder group and its disassembled formula group can reduce serum AngⅡ,ACE2 and Ang(1-7)levels and regulate the over expression of Ang-Ⅱ,Ang(1-7)and ACE2.2.The mRNA and protein expression of ACE2 and Masr in hippocampus and heart:RT-PCR and WB was used to detect the expression of ACE2 and Masr genes and protein in hippocampus and heart.The results showed that modified Chaihu Shugan powder group could up-regulated mRNA and protein expression of ACE2 in the hippocampus and the heart and MasR in the hippocampus,and down regulate mRNA and protein overexpression of MasR in the heart.Conclusion:1.This experiment simulates the clinical etiology of coronary heart disease and depression,and uses the multi factor method of high-fat diet and injection of iso+cums to create a model for 56 days.The rats in the model group have decreased behavioral autonomous activities,exercise ability,desire to explore,hyperlipidemia formation,cardiac morphology and pathological changes,and successfully establish a rat model of myocardial ischemia combined with depression in hyperlipidemia.2.modified Chaihu Shugan powder can prevent and treat myocardial ischemia with depression in hyperlipidemic state by regulating the expression of ACE2 and Masr in hippocampus,exerting anti oxidative stress,protecting brain tissue cells,and improving depression behavior;Through regulating the expression of ACE2 and Masr,anti-inflammatory and antithrombotic effects can be exerted to protect cardiac tissue cells and improve myocardial ischemia.Multi angle regulation of ACE2 ang(1-7)-Masr axis may be one of the mechanisms of prevention and treatment of coronary heart disease with depression.3.The results showed that the expressions of AngⅡ,ACE2,ang(1-7),Masr and other indexes in the disassembled drug groups showed changes with the opposite trend of modeling,indicating that the drugs had specific regulatory effects on the abnormal changes of ACE2 ang(1-7)-Masr axis,and the disassembled drugs had different contributions to regulating the expression changes of these indexes,which had a synergistic effect on the whole formula. |
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