| Purpose:Establish a multi-etiology compound coronary heart disease myocardial infarction rat model based on hypertension combined with hyperlipidemia,through a comprehensive and dynamic observation of the rats’ characterization and behavioral changes at different time periods after surgery,combined with the criteria of TCM syndromes,to explore the syndrome evolution law of the model in different time periods.At the same time,according to microscopic physical and chemical indicators and pathological changes,to reveal the biological basis of the model and explore its correlation with syndrome properties.Methods:1.Spontaneous hypertensive rats(SHR)and Wistar rats at the age of 10 weeks were randomly divided into SHR high-fat group,Wistar high-fat group,SHR general diet group and Wistar general diet group.They were given high-fat feed or normal feed for 4 weeks.Blood pressure and lipids were measured 4 weeks later,and then regrouped into hypertension hyperlipidemia myocardial infarction group(referred to as double-high myocardial infarction group),hypertensive myocardial infarction group,hyperlipidemic myocardial infarction group,myocardial infarction group,hypertension group and sham operation group.The left anterior descending(LAD)ligation was performed in each myocardial infarction group.The hypertension group and the sham operation group were only threaded without ligation,and continued the original feed after operation.The ECG showed 6-8 pathological Q waves,systolic blood pressure(SBP)≥140mmHg,and elevated blood TC or TG or LDL-C to determine the success of the model preparation.Then,the characteristics(activity,sensitivity,irritability,color of nose,lips,tongue,and moisture,etc.)and behavioral changes(including echocardiography,rotation tolerance,stress respiration frequency,grip value,the value of tongue and foot’s r/g/b,etc.)of the rats were collected and analyzed within 1-7 days,7-14 days and 14-28 days after the operation,and combined with the diagnostic criteria of TCM syndromes.Eventually,the syndrome attributes of the model in different time windows were determined.2.Detection of myocardial enzyme indexes(AST,LDH,CK,CKMB),inflammation indexes(TNF-α,IL-1β,IL-6),nerve-Humoral factors(ET,AngⅡ,ALD,BNP,NTpro-BNP),oxidation Stress indicators(MDA,NO,ox-LDL)and cardiac pathological tissues.The correlation analysis was carried out with the syndrome indexes.Results:1.The electrocardiogram of the rats in the double high myocardial infarction group showed 6-8 pathological Q waves in the 3 days after surgery,indicating that the myocardial infarction model was successfully prepared.The blood pressure and blood lipid of the rats in the double-high myocardial infarction group were tested at three time points of postoperative 7d,14d,and 28d.The results showed that the SBP of the rats at each time point was greater than 140mmHg,and the blood lipid levels TG and LDL-C were higher than the hypertensive group.All of them were significantly increased(P<0.01),indicating that the preparation of rats with hypertension and hyperlipidemia combined with myocardial infarction was successful.2.Throughout the observation period,most of the hypertensive group rats had some characteristics with high irritability scores,short rotation tolerance time,increased tongue r-value,yellow urine and dry stools.Combined with the diagnostic criteria of traditional Chinese medicine syndromes,they were judged to be liver-fire hyperactivity syndrome.In the double-high myocardial infarction group,hypertensive myocardial infarction group,high-fat myocardial infarction group,and myocardial infarction group,most of these groups’rats appeared the following symptoms:the stress breathing frequency increased,the grasping force value decreased,the echocardiography EF value decreased,and the tongue-surface and plantar r value decreased,g/b value increased and so on.Combined with the diagnostic criteria of TCM syndromes,indicated the syndrome of qi deficiency and blood stasis appeared.In addition,within 1-7 days after operation,rats in the hypertensive myocardial infarction group and the double-high myocardial infarction group also appeared high irritability scores,short rotation tolerance time,yellow urine,and dry stools,which similar to hypertensive group rats,indicating a syndrome with liver-fire hyperactivity.Within 7-28 days after surgery,the rats in the double-high myocardial infarction group and the hyperlipidemic myocardial infarction group appear to food intake reduced,weight gain,and Lee’s index increased,combined with the increase in blood lipid levels,determine the performance of these two groups’rats within 7-28 days after surgery appeared phlegm turbidity syndrome.3.(1)Myocardial injury:Compared with the hypertensive group,the content of myocardial enzymes(LDH,CK,CK-MB)in the double-height myocardial infarction group increased significantly from 7 to 28 days after surgery(P<0.01),and the AST increased significantly from 7 to 14 days after surgery(P<0.05,P<0.01),The trend of hypertensive myocardial infarction group was the same as that of double height myocardial infarction group,while the increase was slightly lower than that of double height myocardial infarction group.The myocardial enzyme index of the high-fat myocardial infarction group increased from 7 to 28 days after the operation(P<0.05,P<0.01).In the myocardial infarction group,LDH and CK contents increased significantly in the whole observation period after surgery(P<0.05,P<0.01).The results of Masson staining showed that the degree of fibrosis in each model of myocardial infarction gradually increased,the collagen volume fraction was significantly increased(P<0.01),the heart infarction area was significantly increased(P<0.01),the heart/body weight ratio was significantly increased(P<0.05).(2)Inflammatory reaction has been throughout the whole process.Compared with the hypertensive group,IL-6,IL-1β,TNF-α in the double-high myocardial infarction group and the hypertensive myocardial infarction group have significantly increased(P<0.05,P<0.01).And IL-6 in the double-high myocardial infarction group was significantly higher than that in the other groups at 28 days after surgery(P<0.05,P<0.01).The trend of IL-1β,TNF-α in the high-fat myocardial infarction group and the myocardial infarction group was consistent with that in the hypertensive myocardial infarction group,while the IL-6 level in the high-fat myocardial infarction group and the hypertensive myocardial infarction group showed a continuous decline from 7 to 28 days after the operation(P<0.01).(3)Neural hormone:The double-high myocardial infarction group and the hypertensive myocardial infarction group mainly showed that,compared with the hypertension group,ET,ALD,BNP and NT-proBNP increased signilficantly during the entire observation period(P<0.05,P<0.01),and the significant increase of AngⅡ within 7-14 days after operation(P<0.05,P<0.01).The results of BNP and NT-proBNP increased significantly 7-28 days after operation compared with the sham group(P<0.01).The myocardial infarction group showed a significant increase in ET,ALD and NT-proBNP in 7-28 days after surgery(P<0.05,P<0.01).At the same time,the BNP level of the double-height myocardial infarction group was significantly higher than that of the other groups 7 days after surgery(P<0.05,P<0.01)(4)Oxidative stress:Compared with the hypertension group,the MDA and NO in the double-high myocardial infarction group were significantly increased at 7-28 days postoperatively(P<0.01),while ox-LDL decreased significantly at 7 days postoperatively(P<0.01),and showed an upward trend 14-28 days after surgery.In the hypertensive myocardial infarction group,the trend of MDA and NO at 7-28 days after operation was the same as that in the double-high myocardial infarction group.Ox-LDL increased significantly only after 14 days(P<0.01).In the high-fat myocardial infarction group,the content of NO at 7 and 28 days after operation was significantly higher than that in the sham operation group(P<0.01),and at 7-14 days after MDA was significantly higher than that in the sham operation group(P<0.01),Ox-LDL was significantly increased only at 28 days after operation(P<0.01).The trend of NO and MDA in the myocardial infarction group was the same as that in the high-fat myocardial infarction group,and ox-LDL increased significantly during the entire observation period(P<0.01).At the same time,compared with the other groups,the NO content in the double-high myocardial infarction group increased significantly(P<0.01)and the ox-LDL content decreased significantly(P<0.05)at the 7 days after surgery.4.A total of 24 indicators participated in the correlation analysis,23 of which are related to the representative indicators of syndromes.Among them,there are 11,14,13,16 related to liver-fire hyperactivity syndrome,qi deficiency syndrome,blood stasis syndrome,and phlegm turbidity syndrome.There are only four related to all the four syndrome with AST,CKMB,ET,and IL-1β.Conclusion:1.The animal model of hypertension-hyperlipidemia combined with coronary heart disease myocardial infarction manifests as liver-fire hyperactivity syndrome and syndrome of qi deficiency and blood stasis within 1-7 days after operation,and evolves into syndrome ofqi deficiency,blood stasis and phlegm turbidity from 7 to 28 days.Rats in the hypertensive myocardial infarction group appeared liver-fire hyperactivity and qi deficiency and blood stasis syndrome 1-7 days after operation,and showed qi deficiency and blood stasis syndrome 7-28 days after operation.Hyperlipidemic myocardial infarction rats showed qi deficiency and blood stasis syndrome 1-7 days after operation,and 7-28 postoperative showed qi deficiency,blood stasis and phlegm syndrome.The rats in the myocardial infarction group showed qi deficiency and blood stasis syndrome during the whole observation period after operation.Rats in the hypertensive myocardial infarction group always showed signs of hyperactivity of liver-fire hyperactivity syndrome.2.The changes of myocardial enzymes and pathological tissues in this model showed that the degree of myocardial injury was greater than that of the other myocardial infarction groups.Inflammatory reaction has been throughout the whole process,with the most significant change in IL-6.The changes of nerve-humoral factors show that the difference between the high-fat myocardial infarction group and the myocardial infarction group is more obvious.Oxidative stress had a significant effect on each myocardial infarction group,among which the changes of NO and ox-LDL could be the prominent characteristics of the double-high myocardial infarction group.There are 11,14,13,and 16 indicators involved in the formation of liver-fire hyperactivity,qi deficiency,blood stasis,and phlegm turbidity syndrome,and only 4 of them are related to the four syndromes,namely AST,CKMB,ET,IL-1β. |
