| BackgroundAlthough there are many kinds of treatment methods and medications for acute myocardial infarction,and their development and renewal are rapid,the mortality and clinical recurrence rate caused by the disease are still very high.The patients who survive after acute myocardial infarction are further deteriorated and develop to the degree of heart failure.Very large,clinical treatments do not completely resolve many of the patient's symptoms and signs,suggesting that current treatments miss one or more of the pathological mechanisms.The rapid inflammatory response induced by myocardial infarction is an important cause of the initiation of apoptotic signaling pathways in cardiomyocytes in vitro and in vivo,and it is inextricably linked with myocardial fibrosis and the promotion of ventricular remodeling.Two to three hours after myocardial infarction,the ischemic and hypoxic cardiomyocytes and tissues can attract many monocytes in the peripheral circulation to accumulate to the ischemic site,under the stimulation of the infarct tissue microenvironment and after myocardial infarction itself.Under the regulation of mechanism,the differentiation of monocytes into subtype macrophages with different functions plays a different role in ischemic and hypoxic myocardial tissue cells.Toll-like receptors,which are located on the surface of immune cells including macrophages,activate the inflammatory response after myocardial infarction through the TLR4/NF-kB signaling pathway.The Yiqi Huoxue Drug mainly include Huangqi,Chinese angelica,ginseng,Chuanxiong and Panax notoginseng.They are based on the theoretical basis of traditional Chinese medicine theory of blood and blood.Myocardial injury and the effectiveness of myocardial infarction,its role is mainly Yixin Qi,promote blood flow and dredge veins,can significantly improve or even eliminate the symptoms of myocardial infarction patients with pain,chest tightness,shortness of breath and other symptoms,the research group has also been in the previous study It has been proved that Yiqi Huoxue Drug can improve cardiac ejection function in rats with myocardial infarction,improve myocardial damage after myocardial infarction in rats,and reduce serum inflammatory cytokine secretion.PurposeThe experimental subject of this study was to investigate the mechanism of Yiqi Huoxue Drug for improving cardiac function after myocardial infarction by studying the changes of macrophages and related inflammatory factors after myocardial infarction in rats.MethodIn this experiment,the animal model of acute myocardial infarction was established by the method of ligation of the left anterior descending coronary artery in a male SD rat.The MI models were randomly divided into model group,perindopril group and Yiqi Huoxue drug group.In three.groups,a sham group with only puncture and no ligation was set up.There were a total of four groups.The rats were given intragastric administration on the second day after the operation.Changes of each index at two time points after intragastric administration for 7 days and 28 days were observed.Ultrasound and HE staining were used to detect the heart of acute myocardial infarction rats at two time points.Functional structure and pathological changes,overall assessment of cardiac function in rats with myocardial infarction;immunohistochemical detection of CD68,CD86,CD 163,to detect macrophage infiltration and typing changes in the infarct marginal zone,explore the Yiqi activating blood drugs The effect of modulation;The expression of M1,M2,TLR4 and NF-kB protein was detected by Western blot,explored the possible mechanism of inflammatory cytokines modulation by Yiqi Huoxue drug;Serum MCP-1,TNF-a,IL were detected by enzyme-linked immunosorbent assay.-Levels of 6? and IL-10.Results The experimental study was divided into 4 partsIn experiment 1,in order to evaluate the changes of cardiac function in myocardial infarction rats after drug intervention,cardiac ultrasound and HE pathological staining were used for evaluation.The results of HE staining showed that the myocardial cells in the sham-operated group were well-arranged,normal in size and shape,clear in structure,normal in nucleus morphology,and no hypertrophy of cardiomyocytes;the volume of myocardial cells in the model group was significantly increased,and the number of normal cells was significantly reduced.Disorders were arranged and inflammatory cells infiltrated.Each drug group improved compared with the model group.The effects of Yiqi Huoxue Drug on cardiac ultrasound in acute myocardial infarction rats were as follows:compared with sham-operated rats,the left ventricular end-systolic diameter(LVIDs)and left ventricular end-diastolic dimension(LVIDd)of model rats were significantly increased.Left ventricular diastolic wall thickness(LVPWd),left ventricular systolic wall thickness(LVPWs)decreased,left ventricular ejection fraction(LVEF)and left ventricular short axis rate(LVFS)decreased significantly.At 7 days,compared with the model group,the LVIDd and LVIDs of each drug group decreased slightly,but there was no statistical difference(P>0.05),LVPWd,LVPWs had no significant changes,no statistical difference,LVEF,LVFS were increased,including Perindopril increased more significantly,with statistical differences.At 28 days,compared with the model group,the LVIDd and LVIDs of each drug group were significantly reduced,but although the LVIDd of the Qi Qihuoxue drug was reduced,there was no significant difference(P>0.05).The LVPWd and LVPWs were significantly increased,but the Qi Although the LVPWd of the blood-activating drug was reduced without statistical difference,the LVEF and LVFS were significantly increased.Through the above results,it can be found that the left ventricular diastolic and systolic function is significantly reduced after acute myocardial infarction in rats,and after treatment with Yiqi Huoxue,it can improve the left ventricular systolic and diastolic function and reduce the degree of left ventricular dilatation.The recovery of heart function after myocardial infarction plays an active role.In experiment 2,in order to observe macrophage infiltration in the marginal infarct area,immunohistochemical staining was performed.The cells under the microscope were stained with brown-yellow cells as positive cells,and 10 cells in each section were selected for positive cell counts.Finally,the mean value was selected.Immunohistochemical staining of the myocardial infarct marginal tissue showed that there were fewer or no counts of CD68-positive macrophages,CD86-positive macrophages,and CD 163-positive macrophages in the sham-operated group on days 7 and 28,the model group and the sham operation.The number of cells in each positive macrophage group increased significantly compared with the group.At 7 days,the counts of CD68-positive macrophages decreased in the drug-treated group compared with the model group(P<0.05);the CD163-positive macrophage counts in the drug-treated group increased,with statistical significance(P<0.05).The number of CD86-positive macrophages in each drug group was significantly decreased compared with the model group,with statistical significance(P<0.01).At 28 days,the counts of CD68,CD86,and CD 163 positive macrophages in each drug group were significantly lower than those in the model group,and the difference was statistically significant(P<0.01).The results showed that Yiqi Huoxue Drug can reduce the infiltration of macrophages in the border zone of myocardial infarction,reduce the secretion of Ml macrophages,promote the secretion of M2 giant cells,reduce the inflammatory response,and thus reduce myocardial fibrosis and ventricular Refactoring.In experiment 3,in order to further explore the effect of Yiqi Huoxue Pill on the expression of M1 and M2 macrophages in the myocardial tissue of infarct marginal zone in rats with myocardial infarction,and whether the secretion of inflammatory factors is related to the activation of TLR4/NF-kB,through protein immunization.Western blotting was used to determine the protein content in tissues.At 7 days and 28 days,compared with sham operation,the levels of CD86,CD163,TLR4 and NF-kB in the infarct marginal zone of the model group were significantly increased,and the difference was statistically significant(P<0.01).At 7 days,compeared with the model group,the CD86 content in the infarct marginal myocardial tissue of the medication group decreased,and the CD 163 content increased.The TLR4 and NF-kB content decreased,and the difference was statistically significant(P<0.05).At the 28th day,the CD86 content in myocardial tissue in the infarct marginal area of the medication group decreased more significantly(P<0.01),the CD86 content further decreased,and the CD163 content increased significantly.The results were statistically significant(P<0.05).TLR4 and NF-kB decrease in the content was significant and the results were statistically significant(P<0.01).The results showed that Yiqi Huoxue Drug can reduce M1 macrophages and reduce the protein content of TLR4 and NF-k B,thereby reducing the secretion of inflammatory factors and reducing myocardial injury.The results are consistent with the results of immunohistochemistry.In experiment 4,the changes of the corresponding functional indexes in serum were detected by enzyme-linked immunosorbent assay.The correlation factors of interleukin-6(IL-6),tumor necrosis factor-a(TNF-?),monocyte chemoattractant protein-1(MCP-1)and interleukin-10 were detected in each group.At 7 days,serum IL-6 and TNF-a in the sham-operated group were significantly higher than those in the sham-operated group(P<0.01),and each drug group was decreased.Statistical significance(P<0.01).Compared with the sham group,the serum MCP-1 level in the model group increased significantly(P<0.01);the MCP-1 level in the serum of the Yiqi Huoxue Pill group and the perindopril group decreased.Obviously,there was a significant difference(P<0.01).The serum IL-10 in the model group was significantly lower than other groups,with significant differences(P<0.01).At 28 days,the serum levels of IL-6,TNF-a,MCP-1,and IL-10 in the model group were significantly different from those in the sham group(P<0,01 or P<0.05);Compared with the model group,serum IL-6,TNF-?,and MCP-1 were all significantly reduced(P<0.05),while IL-10 was significantly increased(P<0.01).).The results showed that Yiqi Huoxue Drug can down-regulate the levels of inflammatory cytokines in rat serum after myocardial infarction and reduce the production of MCP-1,thereby reducing the recruitment of macrophages in myocardial infarction sites,alleviating myocardial inflammatory responses,and improving myocardial function.Conclusion1L By building a rat model of acute myocardial infarction,it was verified that Yiqi Huoxue Pill can effectively improve the heart structure and function of myocardial infarction rats and play a positive role in the recovery of cardiac function in rats after acute myocardial infarction.2.Yiqihuoxue drug may reduce macrophage infiltration in the infarct marginal zone,regulate the balance of Ml and M2 macrophages,promote the secretion of M2 macrophages,reduce TLR4,NF-kB protein production and reduce myocardial infarction The inflammatory response in the border zone of the rat infarct reduced myocardial injury,reduced myocardial infarct size,and improved myocardial structure and function.3.By detecting the serum levels of IL-6,TNF-a,MCP-1 and IL-10 in rats with myocardial infarction,we further confirmed that Yiqi Huoxue Drug can reduce the inflammatory response in the infarct margin of infarcted rats and improve Heart function. |
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