| Objective:Based on the treatment of acute myocardial infarction by acupuncture at Neiguan point,this study explored the initiation mechanism of acupuncture effect mediated by local adenosine A1 receptor in the point area.Method:1 Effect of acupuncture at Neiguan point on AMI ratsHealthy adult SD rats were selected,six in the sham operation group,other rats were ligated with LAD(Left Anterior Descending coronary artery)to replicate AMI(Acute Myocardial Infarction)model,after modeling,the rats were randomly divided into model group and model needle group,with 6 rats in each group.Immediately after modeling,the ECG of rats was detected by physiological signal recording system,and the success of modeling was preliminarily evaluated by ST segment arch back elevation of 0.2mv.On the first day after modeling,LVEF(Left Ventricular Ejection Fraction)and LVFS(Left Ventricular Fractional Shortening)were measured by small animal ultrasound.On the second day after modeling,the model acupuncture group was treated with bilateral hand acupuncture at Neiguan point once a day for 7 days;The sham operation group and the model group were only fixed with the same binding as the model needle group without hand acupuncture;After 7 days of treatment,LVEF and LVFS of rats in each group were detected,and the myocardial infarction area was detected by TTC staining to evaluate the therapeutic effect of acupuncture on AMI.2 Experimental Study on the effect of acupuncture at Neiguan point on local adenosine A1 receptor and fibroblastsOn the basis of Experiment 1,the local tissues of rat acupoints in each group were further taken,and the protein and gene contents of four adenosine receptors in acupoints were detected by WB(Western blotting)and RT q PCR(Real-time Quantitative PCR Detection System),immunofluorescence staining was used to detect the number of positive expression of A1R(Adenosine A1 receptor),fibroblasts and fibroblast activating protein(FAP)in acupoints,ELISA(Enzyme linked immunosorbent assay)was used to detect the changes of cytokines(IL-6、TNF-α)related to fibroblasts in acupoints,so as to evaluate the effects of acupuncture on A1 R,fibroblasts and fibroblast related cytokines(IL-6、TNF-α)in acupoints.3 Effect of regulating adenosine A1 receptor on acupuncture treatment of AMI rats and its effect on fibroblastsHealthy adult SD rats were selected.there were 6 rats in the sham operation group and the other rats copied the AMI model,after modeling,the rats were randomly divided into model group,model needle group,model needle + DMSO group(control solution),model + CCPA group(A1R agonist)and model needle + DPCPX group(A1R antagonist),with 6 rats in each group.Immediately after modeling,the ECG of rats was detected by physiological signal recording system,and the success of modeling was preliminarily evaluated by ST segment arch back elevation.On the first day after modeling,LVEF and LVFS of rats in each group were detected by small animal ultrasound.On the second day after modeling,the model needle group was treated with acupuncture at both Neiguan points,the model needle + DMSO group and the model needle + DPCPX group were injected with 20 μl of DMSO and DPCPX respectively at Neiguan point half an hour before acupuncture,the model + CCPA group was injected with 20μl of CCPA at Neiguan point every day,the model group was not given any acupuncture and drug intervention.After 7 days of treatment,LVEF and LVFS of rats in each group were detected by small animal ultrasound,and myocardial infarction area was detected by TTC staining,so as to clarify the effect of regulating local A1 R function of acupoint area on acupuncture effect.The number of fibroblasts and activated protein(FAP)in local acupoints were detected by immunofluorescence staining,and the local cytokines(IL-6,TNF-α)in acupoints were detected by(IL-6,TNF-α)to determine the effect of regulating the local A1 R function of acupoint area on fibroblasts and related cytokines.Result:1.Acupuncture at Neiguan point can improve cardiac function and reduce myocardial infarction area in AMI ratsThe AMI model was prepared by ligating LDH,the ECG showed that the ST segment of rats after modeling was significantly higher than that before modeling.The results of cardiac ultrasound showed that compared with the sham operation group,LVEF and LVFS decreased significantly 24 hours after modeling,indicating that the modeling was successful;after 7 days of acupuncture intervention,it can increase LVEF and LVFS and reduce myocardial infarction area in AMI rats(P < 0.01),suggesting that acupuncture at Neiguan point can improve cardiac function and reduce myocardial infarction area in rats.2.Acupuncture at Neiguan point increased the expression of A1 R,the number of fibroblasts and the content of IL-6,TNF-α increased(1)After 7 days of intervention,WB and Q-PCR detection showed that the A1 R protein content and gene content in the model acupuncture group were higher than those in the model group,but there was no significant difference(P > 0.05);immunofluorescence staining showed that the positive expression of A1 R in acupoint area of model needle group was higher than that of model group(P < 0.01),and A1 R in acupoint area was mainly expressed on fibroblasts.(2)Compared with the model group,the number of fibroblasts and the expression of activated protein(FAP)in the acupoint area of the model needle group increased,with statistical difference(P < 0.01).(3)Compared with the model group,the content of cytokine(IL-6、TNF-α)in the acupoint area of the model acupuncture group was significantly higher(P < 0.01).3.Regulating the local A1 R function of acupoint area can affect the effect of acupuncture on AMIAfter 7 days of intervention,compared with the model group,the model acupuncture group could improve the cardiac function of the AMI rats,increase the LVEF and LVFS of the rats,and reduce the myocardial infarction area(P<0.01),suggesting that acupuncture at Neiguan acupoints could improve the cardiac function of the AMI rats.Compared with the die-needle group,the cardiac function of the die-needle+DMSO group and the model+CCPA group had no significant change(P>0.05),while the cardiac function of the die-needle+DPPCX group decreased(P<0.01),indicating that local injection of acupoints The control solution DMSO did not affect the effect of acupuncture,CCPA(A1R agonist)could simulate the effect of acupuncture,and DPCPX(A1R antagonist)partially blocked the effect of acupuncture on improving cardiac function in AMI rats.4.Regulating the local A1 R function of acupoint area can affect the content of fibroblasts and related cytokines.After 7 days of intervention,compared with the model group,the die-needling group,dieneedle+DMSO group,and model+CCPA group could increase the number of fibroblasts and activated protein(FAP)(P<0.05),while the die-needle+DPPCX group was significantly different from the model group.There was no significant change between the groups(P>0.05);compared with the die-needle group,the number of fibroblasts and the number of activated protein(FAP)in the die-needle+DMSO group and the model+CCPA group had no significant changes(P>0.05),while the the number of fibroblasts and the number of activated protein(FAP)in the needle + DPCPX group decreased(P<0.05).It is suggested that acupuncture at Neiguan or local injection of A1 R agonist CCPA can increase the number of fibroblasts and activated proteins in the acupoint,blocking A1 R can partially inhibit the effect of acupuncture on fibroblasts,indicating that acupuncture the effect on fibroblasts is mediated by A1 R.At the same time,compared with the model group,the levels of IL-6 and TNF-α in the acupoints of the die-needling group,the die-needle+DMSO group,and the model+CCPA group increased(P<0.01),while the die-needle+DPPCX group had no significant change(P>0.05);compared with the die-needle group,the contents of IL-6 and TNF-α in the die-needle+DMSO group and the model+CCPA group had no significant changes(P>0.05),however,the content of IL-6 and TNF-α in the die needle + DPCPX group decreased(P<0.01).It is suggested that acupuncture at Neiguan point or local injection of A1 R agonist CCPA at acupoints can promote the increase of local cytokines(IL-6,TNF-α)in acupoints.ImpactConclusion:1.Acupuncture at Neiguan acupoint is effective in the treatment of AMI,can improve cardiac function and reduce myocardial infarction area in AMI rats.2.Acupuncture can activate adenosine A1 receptor on fibroblasts in acupoint area and cause changes in the number and function of fibroblasts,which may be one of the initiating mechanisms of acupuncture in the treatment of AMI. |
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