摘要
背景:帕金森病不仅影响患者的运动功能,而且对感觉、认知也有严重影响。采用多巴胺类药物替代治疗,只能控制症状,并且副作用较大,临床应用中药经验方抑颤汤治疗帕金森病取得了较好的疗效。目的:观察抑颤汤对帕金森病大鼠模型行为学特征、脑组织儿茶酚胺类物质以及黑质神经细胞的影响,分析其可能的作用机制。设计:随机对照动物实验,检测者单盲评估。单位:解放军总医院中医科。材料:雌性SD大鼠120只,体质量180～200g,将造模成功的70只帕金森病模型大鼠通过数字表法随机分生理盐水组30只和抑颤汤组40只,另取未经造模的大鼠30只作为正常对照组。抑颤汤由山萸肉15g,石菖蒲15g,仙灵脾10g,肉从蓉10g,枸杞子15g,丹参15g,蜈蚣8g等组成,由解放军总医院中药房提供。按传统煎药法文火煎30min滤取药液,以恒温水浴锅浓缩成含生药1.0g/mL汤剂。方法:实验于2001-09/12在解放军总医院动物实验室(洁净级)完成。运用6-羟基多巴诱发法建立帕金森病大鼠模型,观察帕金森病大鼠行为特征。抑颤汤组帕金森病大鼠每千克体质量给药量按成人(60kg)每千克体质量剂量(5mL/kg)的10倍计算,灌胃给药,1次/d。每周称体质量1次,按体质量调整给药量,连续给药8周。生理盐水对照组和正常对照组每日按体质量给予等量生理盐水灌胃。分别在给药前、给药第5,6,7,8周,观察正常组、生理盐水组(用药前称模型组)和抑颤汤组大鼠的旋转行为。采用微透析技术和高效液相色谱法测定各组大鼠脑组织细胞外液中儿茶酚胺类物质含量的变化,光学显微镜20倍、40倍及电子显微镜观察双侧纹状体黑质神经细胞的改变。主要观察指标:观察抑颤汤疗效及其对帕金森病大鼠脑神经介质的影响;光学显微镜及电子显微镜观察双侧纹状体黑质神经细胞的改变,评价抑颤汤的作用。结果:各组实验大鼠共100只,由于灌胃操作失当,抑颤汤组、生理盐水组和正常对照组分别死亡6,5,2只,最终进入结果分析87只。①治疗第5,6,7,8周后抑颤汤组大鼠40min旋转次数均明显低于生理盐水组[(112.34±33.36),(91.16±73.06),(72.05±20.77),(61.63±17.93),(340.90±52.97)次,P<0.01]。②帕金森病大鼠损毁侧脑组织透析液中3,4-二羟基苯酸、高香草酸、多巴胺、5-羟色胺水平明显低于未损毁侧(P<0.05～0.01),治疗后抑颤汤组上述介质含量则明显高于生理盐水对照组(P<0.05～0.01),而未损毁侧各介质的含量3组比较差异均没有显著性意义(P>0.05)。③病理学观察,抑颤汤组大鼠受损侧脑黑质神经细胞计数明显多于生理盐水组,且神经元体积较饱满,结构较清晰,细胞内高尔基氏体、线粒体等接近正常。结论:抑颤汤能减轻帕金森病模型大鼠脑黑质细胞的受损程度,促进其修复,提高脑组织中内源性儿茶酚胺类物质的含量,改善帕金森病大鼠的旋转行为。
BACKGROUND： Parkinson disease （PD） not only influences motor function, also affects sensation and cognition. Treatment with dopamine can only control symptom and has well side effect; however, yichan tang has great effect on treating PD in clinic. OBJECTIVE： To observe the effect of y/chan tang on praxiologieal behavior, eateeholamines substance in cerebral tissue and substantia nigTa nerve cells of PD rats and analyze its effective mechanism. DESIGN： Randomized control animal study and single-blind evaluation. SETTING： Department of Traditional Chinese Medicine of General Hospital of Chinese PLA. MATERIALS： A total of 120 female SD rats weighing 180-200 g were selected in this study. Seventy modeling PD rats were randomly divided into saline group （n=30） and yichan tang group （n=40） according to digital table. Another 30 non-modeling rats were regarded as normal control group. Yichan tang was made of 15 g shanyurou, 15 g Shichangpu, 10 g xianlingpl, 10 g roucongrong, 15 g gouqizi, 15 g danshen and 8 g wugong, etc., and provided by Dispensary of Chinese herb of General Hospital of Chinese PLA. Drug was decoeted for 30 minutes＇ to filter the solution and then concentrated into decoction including 1.0 g/mL raw materials. METHODS： The experiment was carried out at the Animal Laboratory of General Hospital of Chinese PLA from September to December 2001. PD rat model were established with 6-hydroxy-dopa to observe behavior of rats. The administration of rats （body mass/kg） in yichan tang group was 10 times as the administration of the adults （60 kg） （5 g/kg）. Rats were perfused once a day and weighted once a week. The dosage of administration was regulated as the body mass changing, and the administration lasted for 8 weeks. Rats in saline group and normal control group were perfused with the same volume of saline as the criteria of body mass. Rotatory behaviors of rats in normal group, saline group （model group before administration） and yichan tang group were observed before administration and at 5, 6, 7 and 8 weeks after administration. Changes of content of catecholamines in extraeellular fluid of cerebral tissue were measured with micro-dialysis technique and high performance liquid chromatography （HPLC）, meanwhile, changes of substantia nigra nerve cells on both sides of corpus striatum were observed under 20-time mad 40-time optic microscopes. MAIN OUTCOME MEASURES： Effect of yichan tang on medium of cranial nerve of PD rats; changes of substantia nigra nerve cells on both sides of corpus striatum with optic microscope and electron microscope; evaluation of yichan tang. RESULTS： Among 100 rats, 6 rats in yichan tang group died because of failure perfusion, 5 in saline group and 2 in normal control group. Therefore, 87 rats were involved in the final analysis. ① At 5, 6, 7 and 8 weeks after treatment, rotatory times within 40 minutes were all shorter in yichan tang group than those in saline group [（112.34±33.36）2 （91.16±73.06）, （72.05±20.77）, （61.63±17.93）, （340.90±52.97） times, P 〈 0.01]. ② Levels of 3,4-dihydroxybenzene acid, homovanilic acid, dopamine and 5-serotonin on the injured side were lower than those on the uninjured side （P 〈 0.05- 0.01）. After treatment, levels of mediums mentioned above were higher in yichan tang group than those in saline group （P 〈 0.05-0.01）; however, there were no significant differences of mediums ＂on the uninjured side in the three groups （P 〉 0.05）. ③ Pathological observation： Amount of sub- stantia nigra nerve cells on the injured side was more in y/chan tang group than that in saline group; moreover, volume of neurons was full, structure was clear, and values of Golgi complex and mitochondria were close to the normals.
CONCLUSION： Yichan tang can relieve injured degrees of subsiantia nigra nerve cells, facilitate the repair, increase the content of endogenetie eateeholamines in cerebral tissue, and improve rotatory behavior of PD rats.
出处
《中国临床康复》
CSCD
北大核心
2006年第31期168-171,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金(39970913)
军队"十五"医药卫生科研基金面上课题(01MA116)~~
