越鞠丸对慢性应激大鼠海马脑源性神经营养因子的影响

Effect of Yueju Wan on expression of brain-derived neurotrophic factor in hippocampus of rats with chronic stress

目的:观察主治郁证的中药方剂越鞠丸对慢性应激状态下大鼠海马脑源性神经营养因子的影响,并与阿米替林作标准对照。方法:实验于2004-01/02在暨南大学医学院中医药实验室进行。①分组:取40只清洁级SD雄性大鼠随机分为正常组、模型组、阿米替林组及越鞠丸组4组,每组10只。②造模:正常组不加刺激,其余各组应用孤养和长期不可预见性的中等强度刺激(包括4℃冰水中游泳5min、禁水24h、禁食24h、夹尾1min、昼夜颠倒24h、摇晃5min,160Hz,40℃环境5min共7种刺激)21d造成慢性应激失调模型。③给药:造模第8天开始给药,模型组及正常组灌胃给生理盐水10mL/kg,阿米替林组灌胃给以阿米替林0.2g/kg,越鞠丸组灌胃给越鞠丸(川芎、栀子、苍术、神曲、香附各等量)50g/kg,1次/d,连续14d。④观察指标:观察造模后各组大鼠体质量、摄食量、糖水消耗量及行为的变化。治疗完成后第2天所有动物麻醉状态下处死并取材,免疫组织化学染色,比较各组大鼠脑海马脑源性神经营养因子阳性神经元的平均吸光度。结果:40只大鼠全部进入结果分析。①脑源性神经营养因子阳性神经元的平均吸光度:模型组脑海马CA1,CA2,CA3,CA4区均低于正常组(P<0.01),而越鞠丸组及阿米替林组明显高于模型组(P<0.01),与正常组接近。②体质量:造模后模型组显著低于正常组(221.3±15.5),(245.1±12.5)g,P<0.01,越鞠丸组显著高于模型组(239.9±10.6)g,P<0.01。③摄食量:造模后模型组显著低于正常组(8.01±1.10),(10.09±1.88)g/kg,P<0.01,越鞠丸组和阿米替林显著高于模型组(9.88±0.91),(9.57±1.01)g/kg,P<0.01。④糖水消耗量:造模后,模型组的糖水消耗显著低于正常组(7.81±2.97),(12.58±3.11)g,t=4.587,P<0.001,越鞠丸组和阿米替林组糖水消耗明显多于模型组(10.97±2.89),(11.01±3.14)g,P<0.01。⑤行为学变化:造模后抑郁组动物水平运动和垂直运动得分都显著低于对照组(P<0.001);越鞠丸组和阿米替林组与模型组比较,水平运动及垂直运动都有改善(P<0.001),接近正常组水平。结论:越鞠丸对慢性应激大鼠抑郁模型有明显的抗抑郁作用,其抗抑郁作用可能与增加海马脑源性神经营养因子的表达有关。

AIM： To observe the effect of Chinese medicine Yueju Wan, which is mainly used to treat depression, on brain-derived neurotrophic factor in hippoeampus of chronic stress rats, and take amitriptyline as the standard control. METHODS： The experiment was completed in the Laboratory of Traditional Chinese Medicine, Medical College of Jinan University from January to February 2004. ①Grouping： Forty male SD rats of clean degree were randomly divided into 4 groups with 10 rats in each group： normal group, model group, amitriptyline group and Yueju Wan group. ② Model establishment： Rats in the control group had no stress, and those in the other groups were subjected to isolated cultivation and long-term unpredictable stress of middle intensity （including swimming in 4℃ ice water for 5 minutes, forbidden from water for 24 hours, fasted for 24 hours, clipping tail for 1 minute, reverse of day and night for 24 hours, tottering for 5 minutes, keeping in the environment of 160 Hz and 40℃ for 5 minutes） for 21 days to induce model of chronic stress disturbance. ③ Administration： From the 8th day after model establishment, gastric perfusion of saline （10 mL/kg） was given in the model group and normal group, gastric perfusion of amitriptyline （0.2 g/kg） was given in the amitriptyline group, and gastric perfusion of 50 g/kg Yueju Wart （szechwan lovge rhizome, cape jasmine fruit, Chinese atractylodes, medicated leaven and nutgrass galingale rhizome of the same dosage） was given in the Yueju Wan group, once a day for 14 continuous days. ④ The body mass, amount of ingestion, consumption amount of sugar water and change of behavior after model establishment were observed. All the rats were killed under the state of anesthesia on the 2^nd day after treatment for material, immunohistochemical staining was applied to compare the average absorbance of the positive neuron of brain-derived neurotrophic factor. RESULTS： All the 40 rats were involved in the analysis of results. ① The average absorbance of the positive neuron of brain-derived neurotrophic factor： Those in the hippocampal CA1, CA2, CA3 and CA4 regions were all lower in the model group than in the normal group （P 〈 0.01）, but high- er in the Yueju Wart group and amitriptyline group （P 〈 0.01）, and close to the normal group. @ Body mass： After model establishment, it was significantly lower in the model group than in the normal group [（221.3±15.5）, （245.1±12.5） g, P 〈 0.01], significantly higher in the yueju Wan group than in the model group [（239.9±10.6）g, P 〈 0.01]. ③ Amount of ingestion： After model establishment, it was significantly lower in the model group than in the normal group [（8.01±1.10）, （10.09±1.88） g/kg, P 〈 0.01], but significantly higher in the Yueju Wan group and amitriptyline group than in the model group [（9.88±0.91）, （9.57±1.01） g/kg, P 〈 0.01]. ④ Consumption amount of sugar water： After model establishment, it was significantly lower in the model group than in the normal group [（7.81±2.97）, （12.58±3.11） g,t=4.587, P 〈 0.001], but obviously more in the Yueju Wan group and amitriptyline group than in the model group [（10.97±2.89）, （11.01±3.14） g, P 〈 0.01]. ⑤ Changes of behavior： After model establishment, the scores of horizontal and vertical movements were significantly lower in the model group than in the normal group （P 〈 0.001）; As compared with the model group, the horizontal and vertical movements were improved in the Yueju Wan group and amitriptyline group, and close to those in the normal group. CONCLUSION： Yueju Wan has an antidepressive effect in chronic stress rat models of depression, and its antidepressive effect may be associated with the increased expression of brain-derived neurotrophic factor in hippocampus.