针刺对脑梗死大鼠脑微循环灌注状态的定性和定量分析(英文)

Qualitative and quantitative analysis of acupuncture on cerebral microcirculation perfusion state in rats with cerebral infarction

背景:脑梗死发生后直接观察梗死区微血管灌注状态是评价治疗效果的最直接证据。目的:动态观察大鼠脑缺血区流质、流场的变化,定性、定量地分析软脑膜缺血区微血管的状态及其所伴随的血细胞的流速及流态变化。设计:随机对照实验。单位:解放军总医院针灸科。材料:实验于2000-05/09在解放军总医院微循环研究室完成。选用健康雌性2月龄Wistar大鼠120只,单纯随机分为针刺组36只,模型组36只,假手术组36只,正常组12只。前3组又按拟观察时段的要求分3,6,24h组,每组12只。方法:针刺组、模型组动物经戊巴比妥钠腹腔麻醉后,用开颅热凝阻断法行大脑中动脉阻塞术。假手术组经麻醉,开颅等手术过程,但不阻断大脑中动脉。正常组不做任何处理。针刺组于大脑中动脉阻塞术60min后,在人中、内关穴区施加针法:先刺双侧内关,接电针仪,疏密波、频率4～10Hz,强度0.4V,持续刺激5min。继续针刺人中,施雀啄手法强刺激10下。假手术组、模型组动物经抓取、固定过程,但不针刺。采用血管内皮细胞荧光染色及白细胞荧光示踪法,结合显微录像系统和计算机图像分析系统,动态定量地观察了针刺对大脑中动脉阻塞后3,6,24h软脑膜微血管形态、密度、血流速度的影响。主要观察指标:①从显微镜下直接观察各组大鼠不同时段软脑膜微血管的形态、密度、血流状况,以及注入荧光剂后血管内皮细胞形态和周围脑组织状况。②定量测量各组大鼠不同时段梗死区微血管密度、血流速度。结果:120只大鼠均进入结果分析。①模型组各时段微血管内皮细胞着色差,组织渗出荧光多,针刺组明显好于模型组。②软脑膜微血管密度:模型组明显低于正常组。针刺组3,6,24h时段明显高于模型组犤(6.92±0.42),(3.25±0.52)根/视野;(7.61±0.51),(3.68±0.32)根/视野;(8.24±0.72),(5.49±1.2)根/视野,q=5.02～13.84,P<0.01犦。③软脑膜微动物白细胞速度:模型组明显低于正常组。针刺组3,6,24h时段明显高于模型组犤(1193±358),(600±261)μm/s;(1112±267),(517±115)μm/s,(1766±293),(611±291)μm/s,q=4.60～8.28,P<0.01犦。结论:在脑梗死早期,缺血区动、静脉、毛细血管断流现象严重,开放的血管数目少,流速减慢,脑灌注不良。针刺可改善缺血区的灌注状态,使开放的血管数增多,血细胞流态改善。

BACKGROUND： Direct observation of infarcted microvascular perfusion after cerebral infarction is the most immediale evidence in evaluation of therapeutic effects. OBJECTIVE： To observe dynamic changes in flowing quality and field in cerebral ischemic area in rats so as to qualitatively and quantitatively analyze the microvascular state in pia matral encephali ischemic area and the changes of associated leukocytic flowing velocity and flowing morphology. DESIGN： Randomized controlled experiment was designed. SETTING： Department of Acupuncture and Moxibustion of General Hospital Chinese PLA. MATERIALS： The experiment was performed in Research Room of Microcirculation of General Hospital Chinese PLA, in which, 120 female Wistar rats of 2-month old were employed and randomized into acupuncture group （36 rats）, model group （36 rats）, sham-operation group （36 rats） and normal group （12 rats）. The first 3 groups were subdivided in 3 hours, 6 hours and 24 hours groups according to the prescriptive time divisions, 12 rats in each group. METHODS： In acupuncture and model groups, after abdominal anesthesia with pentobarbital sodium, the cranium was opened and the occlusion of meddle cerebral artery was performed in cerebrum with heat-coagulation method. In sham-operation group, anesthesia and cranium opening were performed, but the meddle cerebral artery was not occluded. In normal group, no any management was performed. In acupuncture group, 60 minutes after occlusion of meddle cerebral artery in cerebrum, needling was given on Renzhong （GV 26） and Neiguan （PC 6） with electric stimulation of disperse-dense wave, 4-10 Hz frequency and 0.4 V of intensity, lasting for 5 minutes. Afterwards, pecking-acupuncture technique was done on Renzhong （GV 26） to achieve strong stimulation for 10 seconds. In shamoperation and model groups, the rats were in process of grasping and fixation, but acupuncture was not applied. Vascular endotheliocyte fluorescence staining and leukocyte fluorescence tracing method were applied, in combination with microscopic video system and computer imaging analyzing system, to observe in dynamic and quantitatively the influences of acupuncture on pia matral encephali microvascular morphology, density and blood flowing velocity at 3 hours, 6 hours and 24 hours after cerebral occlusion of meddle cerebral artery. MAIN OUTCOME MEASURES：①Pia matral encephali microvascular morphology, density and blood flowing state at various time divisions observed directly microscopically and vascular endotheliocyte morphology and the state of brain tissue which near the microvascular after infusion with fluorescent reagent in each group.② Microvascular density and blood flow velocity in infarcted area at various time divisions measured in quantity in each group, RESULTS： Totally 120 rats all entered result analysis. ① In model group, microvascular endotheliocytes were colored worse and more fluorescent exudation appeared. Those were better remarkably in acupuncture group compared with model group.② Pia matral encephali microvascular density： That in model group was lower than normal group. That at 3 hours, 6 hours and 24 hours in acupuncture group was higher remarkably than model group [（6.92±0.42）, （3.25±0.52） pces/visual field; （7.61~0.51）, （3.68 ±0.32） pces/visual field; （8.24±0.72）, （5.49 ± 1.2） pces/visual field,q=5.02-13.84, P 〈 0.01]. （~） Pia matral encephali microvascular leukocyte velocity： That in model group was lower remarkably in normalgroup. That at 3 hours, 6 hours and 24 hours in acupuncture group was higher remarkably than model group [（1 193±358）, （600±261） μm/s; （1 112±267）, （517±115） μm/s, （1 766±293）, （611±291） μm/s, q=4.60- 8.28, P 〈 0.01]. CONCLUSION： At early stage of cerebral intaretion, broken blood flow is severe in artery, vein and capillary in isehemie area; the numbers of opened vessels are few, flow rate is slow down and cerebral perfusion is unsatisfactory. Aeupuneture amaliorate the mieroeireulation peffusion state in isehemie area, inereases the numbers of opened vessels and improves flowing morphology of blood cell.