基于“内关心胸谋”的针药结合治疗慢性心力衰竭机制

Mechanism of acupuncture-medicine combination for chronic heart failure based on‘Neiguan acupoint(PC6)can cure the cardiothoracic diseases’

目的:观察内关穴区神经标志物蛋白基因产物9.5(PGP9.5)及内皮黏蛋白(EMCN)变化特征,探析慢性心力衰竭疾病状态、药物/针药结合干预方式影响内关穴的神经生物学机制。方法:60只雄性SD大鼠随机分正常组、模型组、西地兰组及针药结合组,除正常组30只外,其余每组10只。模型组予冠状动脉左前降支结扎,自然发展6周成慢性心力衰竭,不予治疗;西地兰组在模型基础上单次给药0.1 mg/kg,每周6次,持续4周;针药结合组则在成模后行左侧内关穴电针,参数为预电针30 min操作,电针参数为2 Hz,3 mA,每周6次,持续4周,给药同西地兰组。Vonfrey、激光多普勒、远红外温度计观察大鼠内关穴区及心脏状态变化,Western Blot法及免疫荧光法检测内关穴区PGP9.5及EMCN表达,评价慢性心力衰竭疾病状态、干预方式影响内关腧穴变化的关联性。结果:模型组双侧内关穴及心脏体表投影区皮下血流减少(P<0.05);单纯使用西地兰可升高正常大鼠心脏体表投影区皮温及皮下血流,双侧内关穴区仅观察到皮下血流灌注率升高,皮温没有显著变化。而针药结合可诱导正常大鼠两侧穴区及心脏体表投影区域的皮温及微循环灌注显著升高(P<0.05),而双侧三阴交穴区并未察到类似现象;西地兰组及针药结合组均可以显著提升模型大鼠机械痛阈及力竭时间,且针药结合组改变更为显著(P<0.05);针药结合组大鼠心脏组织中PGP9.5(P<0.05)及左侧内关穴区皮肤组织PGP9.5及EMCN表达显著增加,心脏组织中EMCN表达减少(P<0.05),且优于西地兰组(P<0.05)。结论:内关穴局部状态受到慢性心力衰竭疾病状态、不同干预方式等因素影响,且与心功能改善变化趋势一致;内关穴区的PGP9.5及ECMN可能是介导腧穴-靶器官相关联的潜在分子。

Objective:To observe the change characteristics of nerve marker protein gene product 9.5(PGP9.5)and endothelial mucin(EMCN)in Neiguan acupoint(PC6)area,and to explore the neurobiological mechanism of chronic heart failure disease status and drug/needle/drug combination intervention on Neiguan acupoint(PC6)changes.Methods:Sixty male SD rats were randomly divided into normal group,chronic heart failure(CHF)model group,digilanid C group and acupuncture-medicine combination group,except for 30 in the normal group and 10 in each group.In the model group,the left anterior descending branch of coronary artery was ligated,and CHF developed naturally for 6 weeks without treatment.In the digilanid C group,O.1 mg/kg was administered as a single dose six times a week for 4 weeks on a model basis.In the acupuncture-medicine combination group,EA was performed at the left PC6 after the model was formed,with the parameters of pre-EA for 30 min,EA parameters of 2 Hz,3 mA,six times a week for 4 weeks,and the same drugs were administered as in the digilanid C group.Laser Doppler and far-infrared thermometers were used to record the changes in subcutaneous blood fow and temperature in the PC6 area and the projection area of the heart before and after a single intervention in normal rats.After 4 weeks of treatment and intervention,we measured the exercise exhaustion time of each group of rats,and Vonfrey measured the mechanical pain threshold of each group of rats,and observed the changes of PGP9.5 and EMCN expression in the left PC6,to analyze the influence of the disease state of CHF and the mode of intervention on the changes of the PC6.We analyzed the correlation between CHF disease status and intervention methods on the changes of PC6.Results:Subcutaneous blood flow was reduced at the PC6 and the projection area of the heart's body surface in the model group bilaterally(P<0.05).The use of digilanid C alone increased the skin temperature and subcutaneous blood flow in the surface projection area of the heart in normal rats,and only an increase in the subcutaneous perfusion rate was observed in the bilateral PC6,with no significant change in the skin temperature,whereas the combination of acupuncture and drugs induced a significant increase in the skin temperature and microcirculatory perfusion in the acupoints on both sides of the heart and the surface projection area of the heart in normal rats(P<0.05),while a similar phenomenon was not observed in the bilateral Sanyinjiao(SP6)acupoints area.Both the digialnid C group and the acupuncturemedicine combination group could significantly elevate the mechanical pain threshold and force time of the model rats,and the changes were more obvious in the needle-medicine combination group(P<0.05).The expression of PGP9.5 in cardiac tissue(P<0.05)and expressions of PGP9.5 and EMCN in the left PC6 of the rats in the needle-medicine combination group were significantly increased,the expression of EMCN in cardiac tissue was significantly decreased(P<0.05),and they were better than those in the digilanid C group(P<0.05).Conclusion:The local status of PC6 is affected by the disease state of CHF,different intervention methods and other factors,and is consistent with the trend of cardiac function improvement.PGP9.5 and ECMN in PC6 may be the potential molecules that mediate the association between acupoints and target organs.