| Background Cardiovascular disease prevention and treatment in China has achieved initial results,but still faces severe challenges.Cardiovascular diseases account for more than 40%of the total number of deaths each year,making it the primary disease affecting residents' lives.In recent years investigation suggests,the heart rhythm that this kind of sudden death causes by coronary heart disease more is caused by wrong,especially with malign ventricular sex heart rhythm happens after myocardial infarction is most common wrong.Studies have shown that sympathetic remodeling plays an important role in the pathogenesis of arrhythmias.After myocardial infarction,with the occurrence of a large number of inflammatory reactions,a large number of macrophages gather in the damaged myocardium,secreted inflammatory factors further activate the downstream pathway,triggering the repair and regeneration of nerves,thus leading to the occurrence of sympathetic nerve remodeling.In this process,there is a key inflammatory factor,nerve growth factor(NGF).As a member of neurotrophic factor,NGF can enhance the differentiation and development of newborn sympathetic nerves,promote the repair response after sympathetic nerve injury,and is one of the important factors involved in nerve repair after myocardial infarction.Yiqi huoxue prescription is composed of astragalus membranaceus,ginseng,chuanxiong,angelica and panax notoginseng.It is an experienced prescription for the clinical treatment of chest arthralgia by the tutor,and has a good clinical effect in improving myocardial ischemia and arrhythmia.Previous studies have shown that yiqi huoxue recipe can improve the inflammatory response associated with myocardial infarction to some extent,and down-regulate inflammatory factors.NGF,as a key inflammatory factor in sympathetic nerve remodeling,may be an important target for yiqi huoxue recipe to inhibit sympathetic nerve remodeling.Objective The purpose of this study was to study the effect of Yiqihuoxue recipe on NGF,upstream macrophages and downstream related receptors,the molecular mechanism of Yiqihuoxue recipe in regulating nerve remodeling was discussed.Methods In this experiment,healthy SD rats were selected and the classic model of myocardial infarction was adopted.The model rats were randomly divided into three groups according to the electrocardiogram:model group,Yiqihuoxue group,positive drug metoprolol group,and the sham operation group was set as the control group.The sham operation only performed pericardial puncture without ligation.The corresponding drugs were administered to each group 24h after surgery,while distilled water was administered to the control group once a day.In 28 days after 3 days,7 days,as the observation point in time,based on former advanced small animal ultrasound examination,testing rats Left ventricular ejection fraction(LVEF),Left ventricular short axis ratio(LVFS),Left ventricular end-diastolic diameter,(LVIDd)and Left ventricular end systolic diameter(LVIDs).Take heart tissue,part of fixed,fixed with 4%formaldehyde solution for pathological Hematoxylin eosin staining method to detect(Hematoxylin-eosin staining method,HE staining)and Immunohistochemical detection(Immunohistochemical,IHC),to observe the pathological changes of rats,and the sense of nerve regeneration,observe the counting macrophages and CD68 positive expression of NGF plays,and CD68 and NGF plays for immunofluorescence double dye,observation of both total positioning,another part of the tube put cryopreserved cryopreserved immune protein imprinting method to detect(Western Blot,WB),to detect edge area of myocardial infarction model NGF,Tropomyosin receptor kinase A(Tropomyosin receptor kinase A,TrkA),Phosphorylation Tropomyosin receptor kinase A(Phosphorylation Tropomyosin receptor kinase A,The protein expression of TrkA and p75 neurotrophic factor receptor(p75NTR).Results This experiment is an animal experiment,which is divided into three parts.Indexes are selected from different angles to observe the influence of yiqi huoxue recipe on nerve growth factor.Experiment 1:evaluation of the effect of Yiqi Huoxue recipe on cardiac function and nerve remodeling in rats with myocardial infarction Ultrasound results:LVEF and LVFS of the model group at each time point were lower than those of the sham operation group(P<0.01).After 3 days of treatment,compared with the model group,LVEF and LVFS in the metoprolol group increased(P<0.05,P<0.01),while LVEF and LVFS in the Yiqihuoxue group showed no significant difference(P>0.05).After7 days of treatment,LVEF and LVFS increased in the metolol group compared with the model group(P<0.01,P<0.05),while LVEF increased in the Yiqihuoxue group(P<0.05)and LVFS showed no significant difference(P>0.05).After 28 days of treatment,LVEF and LVFS in the model group were lower than those in the sham group(P<0.01).Compared with the model group,LVEF and LVFS were increased in both the metoprolol group(P<0.01,P<0.01)and the Yiqihuoxeu group(P<0.01,P<0.05).There was no significant difference between the two groups.Both cardiac function and ventricular structure were improved in the treatment group.Pathological changes:at 3,7 and 28 days,the myocardial cells around the myocardial infarction of rats in the sham operation group were aligned,with the direction and size being basically the same,and there was a small amount of inflammatory cell infiltration.Normal myocardial cells were rarely seen in the model group,with disordered arrangement,unequal size,disordered direction,wide interstitium,and a large number of inflammatory cells infiltration.Myocardial cells in the group treated with Yiqihuoxue and metoprolol were arranged in a neat order compared with that in the model group,and inflammatory cell infiltration was decreased.In the model group,the myocardial structure of infarction and marginal area was severely damaged,and inflammatory cells were clustered.In the treatment group,the recovery was better on 28 days than on 3 or 7 days.TH sympathetic nerve fibers:a very small number of positive nerve fibers could be seen in the field of view of the sham group 3 days after surgery,and a large number of positive nerve fibers could be seen in the field of view of the model group with chaotic distribution,and the density was significantly higher than that of the sham group(P<0.01).In the Yiqihuoxue group and metoprolol group,a small number of positive nerve fibers were distributed evenly,basically in the same direction as cardiac muscle cells,and the density was significantly lower than that in the model group(P<0.01).7 days after surgery,a very small number of positive nerve fibers were observed in the field of vision of the sham group,and a large number of positive nerve fibers were observed in the field of vision of the model group,with chaotic distribution,and the density was significantly higher than that of the sham group(P<0.01).In the Yiqihuoxue group and metoprolol group,a small number of positive nerve fibers were distributed evenly,basically in the same direction as cardiac muscle cells,and the fiber density was significantly lower than that in the model group(P<0.01).On the 28th day after surgery,a small number of positive nerve fibers were observed in the field of view of the sham group,and a large number of positive nerve fibers were observed in the field of view of the model group,with chaotic distribution,and the density was significantly higher than that of the sham group(P<0.01).In the yiqi huoxue group and metoprolol group,a small number of neatly distributed positive nerve fibers were observed in the visual field,which was more than that in the 7-day group,and the fiber density was significantly lower than that in the model group(P<0.01).This experiment can clearly observe the changes in the overall function of the heart and the corresponding pathological changes,and the prescription for invigorating qi and activating blood has obvious improvement effect.Experiment 2:effect of Yiqihuoxue recipe on cytokines related to inflammatory responseImmunohistochemical expression of CD68:the positive expression level of CD68 in the surrounding tissues of infarct was significantly higher in the three time-point model groups(3,7 and 28 days)than in the sham group(P<0.01).Compared with the model group,the positive expression of CD68 in both metoprolol and Yiqihuoxue groups decreased(P<0.01),and the difference between the two groups was not significant.Immunohistochemical NGF expression:the positive expression level of NGF in the perimyocardial infarction tissues of rats in the three time point model groups(3,7,28)was significantly higher than that in the sham group(P<0.01).Compared with the model group,NGF expression levels in the three time points of Yiqihuoxue and metoprolol treatment groups were decreased(P<0.01),with no significant difference between the two groups.At day 3,there was no significant difference between the Yiqihuoxue groups and the model group(P>0.05),and the difference between the metoprolol group and the model group(P<0.01).The positive expression of NGF was decreased in the groups of 7 days and 28 days of Yiqihuoxue and metoprolol dosage(P<0.01).Fluorescence co-localization:in this experiment,it was observed that after myocardial infarction,macrophages significantly increased and clustered in the damaged area,and the expression of NGF increased with the increase of CD68.Yiqi huoxue recipe can inhibit the secretion and aggregation of macrophages to a certain extent,and also reduce the expression of NGF in macrophages.Experiment 3:effect of Yiqi Huoxue recipe on NGF and its receptorWB detection of NGF expression:NGF protein expression around infarction was significantly higher in the three time point model groups than in the sham group(P<0.01).Three days after surgery,NGF protein expression in rats in each group was observed.The results showed that NGF protein expression around infarction was significantly higher in the model group than in the sham group(P<0.01).Three days after the operation,NGF protein expression in the metoprolol group and the Yiqi huoxue group showed a decreased trend compared with that in the model group,but there was no statistical significance(P>0.05).There was no significant difference between the two groups.7 days after surgery,NGF protein expression in the metoprolol group was lower than that in the model group(P<0.01).NGF protein expression in the lYiqihuoxue group was also lower than that in the model group(P<0.01).28 days after surgery,NGF protein expression in the metoprolol group was significantly lower than that in the model group(P<0.01).NGF protein expression in the Yiqihuoxue group was also lower than that in the model group(P<0.01).There was no significant difference between the groups.WB detection of TrkA and p-trka expression:the protein expression of TrkA in each group was observed.The expression level of p-trka protein in the Yiqihuoxue group was different from that in the model group(p<0.01),and the expression level of TrkA protein was also lower than that in the model group,but there was no statistical significance.Seven days after surgery,TrkA and p-trka protein expressions in the two groups were lower than those in the model group(p<0.01).There was no significant difference between the groups.TrkA and p-trka protein expressions in the metoprolol group were lower than those in the model group 28 days after surgery,with statistical significance(p<0.05,p<0.01).TrkA and p-trka protein expressions in the Yiqihuoxue group were also decreased compared with the model group(p<0.05,p<0.01).There was no significant difference between the two groups.WB detection of P75NTR expression:P75NTR protein expression around infarction was significantly higher in the model group than in the sham group(P<0.01).Three days after the operation,the p75NTR protein expression in the metoprolol group was lower than that in the model group,with statistical significance(P<0.01).P75NTR protein expression in the qi-invigorating and blood-activating group was also decreased compared with that in the model group(P<0.01).Seven days after the operation,the p75NTR protein expression in the metoprolol group was lower than that in the model group,with statistical significance(P<0.01).P75NTR protein expression in the Yiqihuoxue group was also decreased compared with that in the model group(P<0.05).There was no significant difference between the groups.28days after surgery,p75NTR protein expression in both groups was significantly lower than that in the model group(P<0.01).P75NTR protein expression in the Yiqihuoxue group was also decreased compared with that in the model group(P<0.01).There was no significant difference between the groups.Conclusion1.Yiqi Huoxue recipe can improve cardiac function and myocardial ischemia,so as to control the occurrence of nerve remodeling.2.Yiqi Huoxue recipe can can inhibit the inflammatory reaction of macrophages,reduce the aggregation of macrophages in the area around myocardial infarction,and regulate the secretion and expression of NGF by inhibiting the synthesis and secretion of NGF by macrophages.3.Yiqi Huoxue recipe can effectively reduce the expression of NGF and inhibit the phosphorylation of its receptor TrkA,thereby reducing the activation of its receptor,avoiding the activation of the downstream pathway and effectively inhibiting nerve remodeling. |
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