| Objectives:To observe and compare the effects of superficial layer(fascial layer on muscle surface),deep layer(mainly median nerve)of Neiguan and low,medium and high frequency electroacupuncture(2Hz,20Hz,100Hz)on tachyarrhythmia model induced by barium chloride in rabbits.To clarify whether the superficial layer of "Neiguan" has anti arrhythmic effect in rabbits,and to compare which of the six stimulation methods combined with low,medium and high frequency electroacupuncture on the superficial and deep layer has the best stimulation effect on the rabbit arrhythmic model of barium chloride replica,and to further discuss the relationship between the stimulation amount combined with the stimulation parameters of"Neiguan" and the acupuncture effect,so as to provide reference for future research The standardization and objectification of research operation and clinical practice can provide reference for experimental data.Methods:50 healthy male New Zealand white rabbits were randomly divided into blank group,model group,shallow 2Hz group,deep 2Hz group,shallow 20Hz group,deep 20Hz group,shallow 100Hz group and deep 100Hz group.The rabbits in each group were given intraperitoneal injection(IP)of chloral hydrate 10%(0.5g 1kg).After anesthesia(muscle relaxation and loss of tension,hind foot reflex disappeared:there was no contraction reaction when the toes were clamped with tweezers,and the breathing was deep and slow but rhythmic),bl-420n biological signal acquisition and analysis system was connected in the way of standard limb lead II In group A,the rabbits with congenital arrhythmia were monitored continuously until sinus rhythm lasted for more than 300 s,which was regarded as normal.At the 5th minute of ECG monitoring,rabbits in blank group were injected with 1 mL 1kg normal saline via ear edge vein,while rabbits in model group and treatment groups were injected with 0,4%(4 mg/kg)barium chloride via ear edge vein according to body weight of 1 mL/kg.After confirming the success of modeling(QRS complex of continuous tall deformity on oscillograph is the sign of successful modeling),the treatment group was sterilized with 75%alcohol with insulated needle(Wuxi Jiajian,diameter 0.3mm xl3mm,needle tip exposed 3mm,needle body with patent nano coating),and then directly punctured the left "Neiguan" point of the rabbit.The depth of the superficial group was 3 In the deep group,the depth of the needle was 5-8 mm,with the point touching the median nerve to cause the rabbit's left upper limb to produce electric convulsion as the degree;another insulating needle was inserted directly at the non acupoint 3 mm above Neiguan point,with the same depth as the previous insulating needle,and the latter treatment groups were connected with Huatuo sdz-v The negative electrode was connected to Neiguan point,and the positive electrode was connected to non acupoint point.According to groups,electroacupuncture frequency of 2Hz,20Hz and 100Hz,continuous wave,intensity of 1.5mA,lasting for 20min were selected.The blank group and model group did not do any stimulation after injection of normal saline and barium chloride.At the end of the observation,the duration of arrhythmia was calculated according to the oscillograph.The left ventricular anterior wall was stained with hematoxylin eosin and detected by biochemical colorimetry and immunohistochemistry.The morphological changes of myocardial cells,Na+-K+-ATPase activity on myocardial cell membrane and connexin 43 of ventricular muscle were observed Objective to discuss the mechanism of Neiguan point in antagonizing arrhythmia.Results:1 the duration of arrhythmia in each group1)There was no change of ECG in the blank group after injection of 1 mL/kg 0.9%normal saline,while in the model group after injection of 1 mL/kg 0.9%normal saline After 0.4%barium chloride,the ECG showed wide abnormal QRS complex,suggesting that the rabbit arrhythmia model was established,and the duration of arrhythmia in the model group was the longest(P<0.05);there was no statisticaldifference in the duration of arrhythmia between the shallow 2Hz group and the model group;the other groups were shortened in varying degrees,and the difference was statistically significant(P<0.05).2)Among the treatment groups,the duration of arrhythmia in shallow 2Hz group was the longest(P<0.01),and that in deep 20Hz group was the shortest(P<0.05).There was no significant difference in the duration of arrhythmia among 2Hz deep group,20Hz shallow group and 100Hz deep group(P<0.05);among the groups,20Hz deep group<100Hz deep group/2Hz deep group/20Hz shallow group<100Hz shallow group<2Hz shallow group2 Changes of myocardial morphology of rabbits in each group1)The myocardial fibers in the blank group were observed in the long band,with 1-2 nuclei,oval or rectangular shape,located in the center of the cells.The staining of perinuclear was light.Myofibrils were arranged in order,and the periodic striations were clear.Myofibrils were located around the periphery,and the cytoplasm was light stained,containing lipofuscin,and there were regular intercalation disk connections between cells.2)In the model group,the myofibers were disordered,eosinophilic,red blood cells exuded in the stroma,intercalated disc thickened,and a large number of vacuoles and nuclei enlarged.3)The myocardial morphology of each treatment group showed pathological changes to different degrees,among which the deep 20Hz group was lighter.3 Changes of Na+-K+-ATP(also known as Na+-K+-ATP pump)enzyme on myocardial cell membrane of rabbits in three groups1)Compared with the blank group,the value of Na+-K+-ATPase in the model group was lower(P<0.05).2)Compared with the model group,there was no significant difference between the superficial group of Neiguan(2Hz,100Hz)and the model group(P>0.05).3)There was no significant difference among the effective treatment groups(P>0.05).4 Changes of connexin 43 relative expression in rabbit ventricular myocytes of four groups1)Compared with the blank group,the relative expression of connexin 43 protein in the model group was significantly lower(P<0.05).2)Compared with the model group,except that there was no statistical difference between the 2Hz shallow group and the model group(P>0.05),the other treatment groups were all improved(P<0.05),and the 20Hz deep group was the most obvious(P=0.018).3)Compared with each treatment group,the relative expression of Cx43 from high to low was 20Hz deep group>100Hz deep group>2Hz deep group>20Hz shallow group>100Hz shallow group>2Hz shallow group.Conclusions:The results showed that:1 Stimulation of the superficial and deep layers of"Neiguan" could shorten the duration of arrhythmia in the model of antiarrhythmic disease induced by barium chloride,and the effect of deep stimulation of"Neiguan" was slightly better than that of shallow stimulation at 2Hz,20Hz and 100Hz.2 In terms of the relationship between frequency and stimulation effect,20Hz electroacupuncture frequency is the best stimulation frequency in the superficial and deep layers of acupoints,2Hz low frequency is ineffective when the stimulation amount in the superficial layer of acupoints does not reach the treatment threshold;100Hz high frequency is effective in the superficial layer,but less than 20Hz;the deep layer also presents the same rule,but 2Hz deep stimulation effect is stronger than the shallow layer,which can play an anti arrhythmic role.3 There is interaction between acupoint level and frequency.The stimulation effect of shallow and deep level of Neiguan acupoint and different frequency is comprehensive,20Hz deep level group>2Hz/100Hz deep level group/20Hz shallow level group>100Hz shallow level group>2Hz shallow level group,among which the shallow level of 20Hz is better than the deep level of 100Hz,which indicates that the stimulation effect of shallow level of acupoint can be equivalent to that of deep level of acupoint,while the deep level of acupoint is not And or more than the appropriate frequency will cause the reduction of stimulation effect.4 In this experiment,the stimulation effect of Electroacupuncture with 20Hz frequency acting on the deep layer of Neiguan point is the best in antagonizing the arrhythmia induced by barium chloride in rabbits,and its mechanism may be related to improving the function of sodium potassium pump and increasing the number of connexin channel proteins in ventricular muscle. |
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