| 1 Objective(1)Observation the electro-acupuncture stimulate the heart meridians simultaneously to effect the sympathetic nerve and vagus nerve in acute myocardial ischemia(AMI)rat,in order to discuss the heart meridians section improve the coordinate regulation effect by heart automatic nervous in AMI on electro-acupuncture;(2)Observation the paraventricular nucleus(PVN)neuron discharge,the sympathetic nerve discharge and the changes of hemodynamic index simultaneously,then carries on the correlation analysis,in order to explore the hippocampus-PVN-sympathetic neural pathways in the role of acupuncture heart meridian against myocardial ischemia;(3)Observation the nucleus tractus solitarii(NTS)neuron discharge,the vagus nerve discharge and the changes of hemodynamic index simultaneously,then carries on the correlation analysis,in order to explore the hippocampus-NTS-vagus neural pathways in the role of acupuncture heart meridian against myocardial ischemia.2 Methods(1)Effects of Electro-acupuncture of Heart Meridians on the sympathetic and vagus Nervous discharge in Acute Myocardial Ischemia Rats.According to random number table method,the SD rats were randomly divided into sham group,model group and heart meridian group,10 rats in each group.Model group and heart meridian group are reference the operation way of Guo Yao,according the left anterior descending coronary artery ligation method to copy the model of acute myocardial ischemia rat.The heart meridian group adopts the electro-acupuncture to stimulate the heart meridian section: " Shen Men(HT-7)-Tong Li(HT-5)".In the heart meridian group,using three ?0.30×25mm acupuncture needles to stab the upper limbs,distance of 2 mm,connecting the electro-acupuncture therapeutic apparatus,stimulation parameters are consistent,current intensity 1 m A,the frequency of 2 Hz,electro-acupuncture continuous 20 min.Using two platinum wire electrode to lead on the left side of the sympathetic and vagus nerve respectively,through the Bio Pac biological signal collection system to record the ECG,ECG J-spot,the sympathetic nerve and vagus nerve's discharge frequency at the same time in 5 min before acupuncture,in 1 min,3 min,5 min and 15 min after acupuncture.Through the Acqknowledge software to analysis each groups the rat heart rate(HR),electrocardiogram(ECG)J point,sympathetic nerve and vagus nerve discharge frequency.(2)Effects of Electro-acupuncture of Heart Meridians on the PVN neuron discharge,the sympathetic nerve discharge and the changes of hemodynamic index in Acute Myocardial Ischemia Rats.According to random number table method,the SD rats were randomly divided into sham group,model group,heart meridian group and lesion hippocampus+heart meridian group,6 rats in each group.Model group,heart meridian group and lesion hippocampus+heart meridian group are reference the operation way of Guo Yao,according the left anterior descending coronary artery ligation method to copy the model of acute myocardial ischemia rat.Lesion hippocampus+heart meridian group adopt the way of chemical damage using the brain trace injection of Kainic Acid,damaged bilateral hippocampus.The heart meridian group and Lesion hippocampus+heart meridian group adopt the electro-acupuncture to stimulate the heart meridian section: " Shen Men(HT-7)-Tong Li(HT-5)".Stimulation parameters are consistent,current intensity 1 m A,the frequency of 2 Hz,electro-acupuncture continuous 30 min,once a day for 3 times.The microelectrode array embedded in the PVN of rats with each groups,use of Plexon in multi-channel signal acquisition system,records of each rat PVN neurons action Potential(Spikes)and the Local Field potentials(LFP),through the Offline Sorter software to cope with the data processing and cluster analysis by use of the original electrical signal(Wide Band,WB),using the Neuro Explorer software to analyze the rat's PVN neurons discharge frequency(spike counts),peak value,max peak value and spectrogram,and taking the sampling sequence to analyze the autocorrelation analysis after electro-acupuncture in each groups;using BIOPAC multilead physiological recording instrument to observe the sympathetic nerve discharge,HR,Mean Arterial Pressure(MAP)and Rate Pressure Product(RPP)in each groups. (3)Effects of Electro-acupuncture of Heart Meridians on the NTS neuron discharge,the vagus nerve discharge and the changes of hemodynamic index in Acute Myocardial Ischemia Rats.According to random number table method,the SD rats were randomly divided into sham group,model group,heart meridian group and lesion hippocampus+heart meridian group,6 rats in each group.Model group,heart meridian group and lesion hippocampus+heart meridian group are reference the operation way of Guo Yao,according the left anterior descending coronary artery ligation method to copy the model of acute myocardial ischemia rat.Lesion hippocampus+heart meridian group adopt the way of chemical damage using the brain trace injection of Kainic Acid,damaged bilateral hippocampus.The heart meridian group and lesion hippocampus+heart meridian group adopt the electro-acupuncture to stimulate the heart meridian section: " Shen Men(HT-7)-Tong Li(HT-5)".Stimulation parameters are consistent,current intensity 1 m A,the frequency of 2 Hz,electro-acupuncture continuous 30 min,once a day for 3 times.The microelectrode array embedded in the NTS of rats with each groups,use of Plexon in multi-channel signal acquisition system,records of each rat NTS neurons action Potential(Spikes)and the Local Field potentials(LFP),through the Offline Sorter software to cope with the data processing and cluster analysis by use of the original electrical signal(Wide Band,WB),using the Neuro Explorer software to analyze the rat's NTS neurons discharge frequency(spike counts,firing rate),peak value,max peak value and spectrogram,and taking the sampling sequence to analyze the interspike intervals(ISI)and autocorrelation analysisafter electro-acupuncture in each groups;using BIOPAC multilead physiological recording instrument to observe the sympathetic nerve discharge,HR,Mean Arterial Pressure(MAP)and Rate Pressure Product(RPP)in each groups.3 Results(1)Before the AMI model copy,the ECG,ECG J-spot,the sympathetic nerve and vagus nerve discharge are basically consistent(P>0.05);after the AMI model copy,compared with the sham operation group,the ECG,the ECG J-spot and the sympathetic nerve discharge were obviously increased in each time point by the model group(P<0.01),the vagus nerve discharge was obviously decreased in each time point by the model group(P<0.01);compared with the model group,the ECG,the ECG J-spot and the sympathetic nerve discharge were obviously decreased in each time point by the heart meridian group(P<0.01),the vagus nerve discharge was obviously increased in each time point by the heart meridian group(P<0.01),in addition,above of each index have present their respective opposite change tendency by 5 min before and after acupuncture(P<0.05,P<0.01).(2)(1)The changes of rat sympathetic nerve,hemodynamic index in each groups: Compared with the sham group,the sympathetic discharge frequency and HR were significantly increased in model group(P<0.01),MAP,RPP significantly reduced(P < 0.01);Compared with the model group,the sympathetic discharge frequency and HR were significantly decreased in heart meridian group(P < 0.01),MAP,RPP significantly increased(P<0.01,P<0.05);Compared with the heart meridian group,the sympathetic discharge frequency and HR were significantly increased in lesion hippocampus+heart meridian group(P<0.01),MAP,RPP significantly reduced(P < 0.01);(2)The PVN neurons discharge cluster analysis in each groups: the sham group has 2 neurons,the model group has 1 neuron,the heart meridian group has 4 neurons and the lesion hippocampus+heart meridian group has 3 neurons;(3)The PVN neurons discharge frequency in each groups: Compared with the sham group,the PVN neurons discharge was significantly increased in model group(P<0.01);Compared with the model group,the PVN neurons discharge was significantly decreased in heart meridian group(P<0.01);Compared with the heart meridian group,the PVN neurons discharge was significantly increased in lesion hippocampus+heart meridian group(P<0.01);(4)The energy of spectrogram from high to low in the order for each groups: the model group,the lesion hippocampus+heart meridian group,the heart meridian group,the sham group;(5)The correlation analysis between the PVN discharge frequency and sympathetic nerve discharge frequency,hemodynamic index in each groups: the PVN total discharge frequency with the sympathetic nerve discharge frequency were positively correlated(P<0.01),and the inter neurons of the PVN with the sympathetic nerve discharge frequency were positively correlated(P<0.01),but the pyramidal cell of the PVN was not associated with the sympathetic nerve discharge frequency(P>0.05);the PVN total discharge frequency were positively correlated with HR(P<0.01)and negatively correlated with MAP,RPP(P<0.01);the inter neurons of the PVN discharge frequency were positively correlated with HR(P<0.01)and negatively correlated with MAP,RPP(P<0.01),but the pyramidal cell of the PVN was not associated with HR,MAP and RPP(P>0.05).(3)(1)The changes of rats vagus nerve and hemodynamic index in each groups: Compared with the sham group,the HR were significantly increased(P<0.01),the vagus discharge frequency and MAP,RPP were significantly reduced in model group(P < 0.01);Compared with the model group,the HR were significantly decreased(P<0.01),the vagus discharge frequency and MAP,RPP significantly increased in heart meridian group(P<0.01,P<0.05);Compared with the heart meridian group,the HR were significantly increased(P<0.01),the vagus discharge frequency and MAP,RPP significantly reduced in lesion hippocampus+heart meridian group(P < 0.01);(2)The NTS neurons discharge cluster analysis in each groups: the sham group has 1 neurons,the model group has 1 neuron,the heart meridian group has 3 neurons and the lesion hippocampus+heart meridian group has 6 neurons;(3)The interspike intervals analysis of the NTS neurons discharge in each groups: The sham group neuron discharge numbers were relatively concentrated and the neurons within the nucleus were relatively stable;the model group neuron discharge numbers were decrease the discrete tendency;the heart meridian group had 2 numbers of neurons discharge relatively concentrated,1 was discrete;lesion hippocampus+heart meridian group inter neuron discharge numbers were mostly discrete,pyramidal cell discharge numbers were irregularly discrete;(4)The NTS neurons discharge frequency in each groups: Compared with the sham group,the NTS neurons discharge was significantly decreased in model group(P<0.01);Compared with the model group,the NTS neurons discharge was significantly increased in heart meridian group(P<0.01);Compared with the heart meridian group,the NTS neurons discharge was significantly increased in lesion hippocampus+heart meridian group(P<0.01);(5)The energy of spectrogram from high to low in the order for each groups: the lesion hippocampus+heart meridian group,the sham group,the heart meridian group,the model group;(6)The correlation analysis between the vagus nerve discharge frequency and hemodynamic index in each groups: the vagus nerve discharge frequency were negatively correlated with HR(P<0.05)and positively correlated with MAP,RPP(P<0.05);(7)The correlation analysis between the NTS discharge frequency and vagus nerve discharge frequency,hemodynamic index in each groups: the NTS total discharge frequency with the vagus nerve discharge frequency were positively correlated(P<0.01),the NTS total discharge frequency were not associated with the HR(P>0.05),and the MAP,RPP(P>0.05);the 3 neurons in heart meridian group,inter neuron C with the vagus nerve discharge frequency were negatively correlated(P<0.05),with the HR were negatively correlated(P<0.05),with the MAP,RPP were negatively correlated(P<0.05,P<0.01);but the inter neuron A and B were not associated with the vagus nerve discharge frequency,the HR,the MAP and RPP(P>0.05);the 6 neurons in the lesion hippocampus+heart meridian group,inter neuron C,D and E with the vagus nerve discharge frequency were positively correlated(P<0.01),with the HR were positively correlated(P<0.01,P<0.05),with the MAP,RPP were positively correlated(P<0.01,P<0.05),but the inter neuron A and B,pyramidal cell F were not associated with the vagus nerve discharge frequency,the HR,the MAP and RPP(P>0.05).4 Conclusion(1)Electro-acupuncture stimulates the heart meridian may promote the discharge frequency of the vagus nerve and reduce the discharge frequency of the sympathetic nerve.This result could reveal the electro-acupuncture intervene the acute myocardial ischemia,this trend may play a role of the coordinate regulation effect to improve the acute myocardial ischemia;(2)Hippocampus-PVN-sympathetic nerve participate in the effect of the electroacupuncture intervene the acute myocardial ischemia;the inter neuron of the PVN is the key neuron that make the PVN take part in the effect of the electro-acupuncture intervene the acute myocardial ischemia;(3)Hippocampus-NTS-vagus nerve participate in the effect of the electroacupuncture intervene the acute myocardial ischemia;the part of the inter neuron in the NTS is the key neuron that make the NTS take part in the effect of the electro-acupuncture intervene the acute myocardial ischemia. |
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