解毒通络方对脑缺血损伤大鼠海马与皮质神经递质变化的影响

Effects of prescription for detoxification and dredging meridian-collaterals on neurotransmitter in hippocampus and cortex of rats with cerebral ischemia

目的:观察解毒通络方对脑缺血大鼠恢复期海马与皮质中单胺类递质和乙酰胆碱含量的影响。方法:实验于2005-05/2006-02在北京中医药大学病理实验室与中国中医科学院西苑中医院药理实验室完成。选择健康SD大鼠150只,采用双侧颈总动脉夹闭结合低血压状态制作脑缺血模型,将造模成功的150只大鼠随机数字表法分为5组,正常组、模型组及解毒通络方低剂量组、中剂量组、高剂量组,各30只。每组又分为造模后4,8周2个时相点,每个时相点15只。各剂量治疗组每天按不同剂量灌服解毒通络口服液(由栀子、丹参、黄芪、天麻等组成),低、中、高剂量分别相当于生药1.85,3.7,7.4g/kg,配置成给药体积为5mL/kg的溶液;模型组、正常组同时灌胃相应体积的洁净水。以改进的柱前、柱后双酶柱结合高效液相电化学方法检测大鼠海马与皮质的乙酰胆碱含量;以高效液相电化学方法检测大鼠海马与皮质的单胺类神经递质及其代谢产物的含量。结果:纳入动物150只,均进入结果分析。①脑缺血后4周模型组大鼠海马去甲肾上腺素水平与正常组比较显著升高[分别为(797.29±60.04),(528.90±149.13)μg/L,P<0.01]、乙酰胆碱水平显著降低[分别为(19.72±10.09),(39.73±22.31)μg/L,P<0.01],皮质去甲肾上腺素、高香草酸水平与正常组比较显著降低[分别为(52.81±21.71),(258.65±125.82)μg/L,P<0.01;(5.22±3.19),(9.02±2.27)μg/L,P<0.05],海马与皮质多巴胺、5-羟色胺及其他代谢物及皮质乙酰胆碱水平均无明显变化。②脑缺血后8周模型组大鼠海马去甲肾上腺素、多巴胺水平与正常组比较显著升高[分别为(346.88±125.98),(110.76±18.56)μg/L,P<0.05;(7.16±3.52),(3.78±0.44)μg/L,P<0.01],5-羟色胺、乙酰胆碱水平显著降低[分别为(79.38±26.47),(127.64±23.03)μg/L,P<0.01;(14.75±5.21),(22.59±6.58)μg/L,P<0.05],皮质去甲肾上腺素水平也显著降低[分别为(57.67±18.30),(133.43±39.15)μg/L,P<0.01],皮质单胺类递质代谢产物及乙酰胆碱水平则无明显变化。解毒通络方可使上述各项指标的绝大部分恢复到接近正常大鼠水平。结论:解毒通络方能够有效促进脑缺血大鼠海马与皮质中单胺类递质和乙酰胆碱代谢紊乱状态的恢复。

AIM： To study the effects of prescription for detoxification and dredging meridian-collaterals on contents of acetylcholine and monoamines in hippocampus and cortex of rats with cerebral ischemia of convalescence stage. METHODS： The experiment was conducted in the Laboratory of Pathology, Beijing University of Traditional Chinese Medicine and Laboratory of Pharmacology, Xiyuan Hospital of China Academy of Chinese Medical Sciences. 150 healthy SD rats were made into models of cerebral ischemia by occluding bilateral carotid arteries combined with hypotension. 150 rats of successful modeling were randomly divided into 5 groups： normal group, model group and high/middle/low dose prescription for detoxiflcation and dredging meridian-collaterals group （high/middle/low dose prescription group） with 30 rats in each group. Each group was subdivided into 2 groups according to 4 and 8 weeks after modeling with 15 rats in each time-point. Rats in the high/middle/low dose prescription groups orally took detoxification and dredging meridian-collaterals fluid at different dosage （composed of cape jasmine fruit, danshen root, radix astragali and gastrodia rhizome etc.）, the volume of which was 5 mL/kg and were prepared by 1.85 g/kg, 3.7 g/kg and 7.4g/kg crude drug respectively） administration was performed daily. Rats in the model group and normal group received gastric perfusion of purified water at the same volume. The level of acetylcholine was determined by integrated method of improved high performance liquid chromatography （HPLC-ECD） and two immobilized enzyme reactors. The contents of monoamine neuretransmitter and its metabolic product in hippocampus and cortex were determined with HPLC-ECD. RESULTS： A total of 150 animals were involved in the analysis of results. ①At 4 weeks after cerebral ischemia, the level of norepinephrine （NE） in hippocampus of rats in the model group was remarkably increased than the normal group [respectively were （797.29±60.04） and （528.90±149.13） μg/L,P 〈 0.01], and level of acetylcholine was significantly decreased [（19.72±10.09）, （39.73±22.31） μg/L, P 〈 0.01], while the level of NE and homovanillic acid （HVA） in cortex decreased significantly [52.81±21.71], （258.65＋125.82） μg/L, P 〈 0.01 ; （5.22±3.19）, （9.02＋9.27） μg/L, P 〈 0.05]. There were no obvious differences in dihydroxyphenyl ethylamine （DA）, 5-HT and other metabolisms in hippocampus and cortex as well as the level of acetylcholine in cortex.②At the 8^th week of cerebral ischemia, the levels of NE and DA in hippocampus of rats in the model group were obviously increased than the normal group [respectively were （346.88 ±125.98）, （110.76±18.56） μg/L, P 〈 0.05; （7.16±3.52）, （3.78±0.44） μg/L, P 〈 0.01], while levels of 5-HT and acetylcholine were decreased significantly [respectively were （79.38±26.47）, （127.64±23.03） μg/L,P 〈 0.01; （14.75±5.21）, （22.59±6.58） μg/L, P 〈 0.05], and level of cortical norepinephrine was greatly decreased [57.67±18.30], （133.43±39.15） μg/L, P 〈 0.01]. However, levels of cortical neui＇otransmitter metabolic products and acetylcholine were not significantly changed. The prescription for detoxification and dredging meridian-collaterals could enable most of above-mentioned indexes to recovery to normal level. CONCLUSION： The metabolic disorders of monoamine neurotransmitter and acetylcholine in hippocampus and cortex of rats with cerebral ischemia can recover to normal levels by prescription for detoxification and dredging meridian-collaterals.