针刺苍白球内侧部对帕金森病模型大鼠脑内抗氧化酶的调节

Adjustment of anti-oxidase by puncturing medial part of globus pallidus of model rats with Parkinson disease

目的:观察针刺苍白球内侧部对帕金森病模型大鼠行为学及纹状体抗氧化酶的影响。方法:①实验于2001-07/12在黑龙江中医药大学针灸生理实验室完成。选用健康Wistar大鼠45只。②选取35只大鼠,应用偏侧纹状体立体定位注射6-羟基多巴胺的方法制备帕金森病模型大鼠,将造模成功的帕金森病模型大鼠18只随机分为2组:模型组9只,治疗组9只。治疗组:借助脑立体定位技术将改制的针灸针垂直刺入帕金森病模型大鼠患侧苍白球内侧部进行针刺治疗,1次/d,连续治疗1周;然后再应用G-6805Ⅱ型电针治疗仪进行电针治疗犤强度1mA左右(以大鼠能耐受且不激怒、嘶叫为标准),频率为100Hz,连续波犦,持续15min,1次/d,连续治疗2周。另取10只大鼠为空白对照组,在大鼠左侧纹状体区注射含0.2%的抗坏血酸的生理盐水5μL。模型组及空白对照组:不再做任何处理,正常饲养3周。③分别于针刺治疗前、针刺后1周、电针后1周、电针后2周后于3组大鼠背部皮下注射阿朴吗啡0.5mg/kg诱发旋转,记录给药后10～40min内各组大鼠的旋转次数。④行为学检测完毕后,按照南京建成生物工程研究所试剂盒说明步骤,取组织匀浆上清液,通过722分光光度仪用化学比色法测定各组大鼠纹状体谷胱甘肽、谷胱甘肽过氧化物酶、超氧化物歧化酶、丙二醛的含量。⑤计量资料差异比较采用t检验。结果:因造模失败脱失17只,进入结果分析28只。①治疗组从治疗后1周开始,旋转次数逐渐减少,针刺治疗后1周,电针治疗后1和2周分别比治疗前减少14.55%,49.50%,67.07%,即随着治疗时间的延长,帕金森病大鼠的行为学改变明显好转。针刺治疗后1周,其旋转次数与治疗前比较,差异不明显(P>0.05);电针治疗后1和2周,旋转次数明显少于治疗前(P<0.01)。②模型组大鼠纹状体丙二醛含量明显高于空白对照组(P<0.01)。治疗组大鼠纹状体丙二醛含量明显低于模型组(P<0.01)。模型组超氧化物歧化酶含量与空白对照组比较,差异不明显(P>0.05)。模型组大鼠谷胱甘肽、谷胱甘肽过氧化物酶含量明显低于空白对照组和治疗组(P<0.05～0.01)。3组大鼠纹状体超氧化物歧化酶含量差异不显著(P>0.05)。结论:针刺苍白球内侧部可明显改善帕金森病模型大鼠的行为学,同时针刺苍白球内侧部可以减轻多巴胺能神经元的脂质过氧化反应,对氧化应激所引起的神经损伤具有保护作用。

AIM： To investigate the influence of puncturing internal globus pallidus （Gpi） on behaviour and auti-dixidase in eaudate putamen （CPu） in model rats with Parkinson disease （PD）. METHODS： ①Experiments were completed at Acupuncture Physiology Laboratory in Heilongjiang University of TCM from July to December 2001. Totally 45 healthy adult Wistar rats was used in the experiments. ② PD animal model was developed by stereotaxic injection of 6-hydroxydopamine （6-OHDA） into the left striatum in 35 adult Wistar rats. The successful models were randomly divided into control and treatment groups with 9 rats in each group. Treatment group： With the assistance of stereotaxic techniques, the modified acupuncture needles were inserted perpendicularly into the diseased GPi of the rat model and treatment was performed once per day lasting one week, electrical stimulation were performed with G-6805 Ⅱelectric acupuncture treatment system （intensity 1 mA, frequency 100 Hz, consecutive wave, the standard was set as tolerance of the animal to the stimulation without in＇itation and bray） one week after the operation lasting two weeks. Ten rats in blank control group were injected into left striatum with 5 μL of 0.2% ascorbic acid saline solution. Model group and blank control group： No treatment was performed and fed normally for three weeks. ③ 0.5 mg/kg apomorphine were injected subcutaneously into the back of the blank control, model control and acupuncture treatment group before the acupuncture, one week after the acupuncture needle insertion, one week and two weeks after electrical acupuncture treatment,10-40 minutes after the apomorphine administration the times of rotation induced by apomorphine were recorded. ④ The animals were sacrificed just after the last behaviour test, brain were taken and tissue homogenate supernatant were prepared for and contents of reduced glutathione （GSH）, glutathione peroxidase （GSH-Px）, superoxide dismutase （SOD） and malondialdehyde （MDA） were detected with ehromatometrlc method by 722 spectrophotometer according to the instruction of the .kit from Nanjing Jiancheng Bioengineering Institute. ⑤ Measurement data were compared with t test. RESULTS： Totally 17 rats were dead from the failure of model development, 28 were finally used for data analysis.① One week after the acupuncture needle insertion, the rotation was observed decreased 14.55% in treatment group compared with model group, 1 week and 2 weeks after the electric acupuncture treatment, the rotation was reduced 49.55% and 67.07% respectively compared with that before treatment （P 〈 0.01）, it indicate the behaviour was improved in rat PD model by continuous treatment with acupuncture, however, the rotation times before the treatment did not change significantly compared with that after 1 week acupuncture treatment （P〉 0.05）. ② MDA content was much higher in model group compared with blank control group （P 〈 0.01）, MDA content decreased significantly in treatment group compared with the model control group （P 〈 0.01）. In model group, there is no significant difference of the SOD content compared with blank control group （P 〉 0.05）; however, GSH, GSH-Px content were much lower in model group compared with treatment group and blank control group （P 〈 0.01-0.05）; there was no significant changes of SOD among the three groups （P 〉 0.05）. CONCLUSION： Acupuncture treatment can significantly improve behavior of rat PD model, and puncturing GPi can reduce lipid peroxidation of DA neurons and protect nerve injury from oxidative stress accordingly.