银杏叶提取物及银杏总内酯对帕金森病大鼠黑质神经元损伤的保护作用

Effects of ginkgo biloba extract and bilobalide on neuronal injury of substantia nigra of rats with Parkinson disease

目的:观察银杏叶提取物及银杏总内酯预防和治疗帕金森病模型大鼠黑质细胞损伤的作用。方法:实验于2004-07在锦州医学院动物实验室完成。选用健康成年SD雄性大鼠80只。①预防性用药实验:选取40只大鼠,随机分为5组,每组8只:银杏叶提取物预防组,按100mg/kg剂量灌胃银杏叶提取物溶液(含黄酮24%,银杏内酯6%,由北京第四制药厂提供,生产批号0310469,5mL/支),银杏总内酯高剂量预防组,按12mg/kg剂量给予银杏总内酯溶液(首都医科大学药物研究所提供,生产批号2003196,2mg/支),银杏总内酯低剂量预防组,按6mg/kg剂量给予银杏总内酯溶液,对照组和模型组灌胃等体积生理盐水。给药1次/d,连续14d,最末次给药1h后,模型组和用药组通过6-羟基多巴脑立体定向注射术建立帕金森病大鼠模型。②治疗性用药实验:于另40只大鼠中选用8只为对照组(灌胃等体积生理盐水)。将其余32只造成帕金森病模型,均造模成功。随机分为4组,每组8只:银杏叶提取物治疗组(按100mg/kg剂量灌胃银杏叶提取物溶液),银杏总内酯高剂量治疗组(按12mg/kg剂量给予银杏总内酯溶液),银杏总内酯低剂量治疗组(按6mg/kg剂量给予银杏总内酯溶液),模型组(灌胃等体积生理盐水)。给药1次/d,连续14d。③预防性给药组于造模后,治疗性给药组于给药14d后将各组大鼠断头取脑,分离双侧纹状体,采用高压液相色谱仪紫外检测器测定多巴胺浓度;严格按购于南京建成生物研究所的试剂盒说明对右侧黑质超氧化物歧化酶、丙二醛含量进行检测。④计量资料差异比较采用t检验。结果:大鼠80只均进入结果分析。①多巴胺含量:银杏叶提取物预防组和银杏总内酯高、低剂量预防组和对照组大鼠患侧纹状体明显高于模型组(P<0.05～0.01);各组健侧纹状体差异不明显(P>0.05);银杏总内酯高剂量治疗组帕金森大鼠患侧纹状体明显高于模型组(P<0.05),对照组大鼠患侧纹状体明显高于模型组(P<0.01);各组健侧纹状体差异不明显(P>0.05)。②右侧黑质丙二醛含量:各用药预防组和各用药治疗组及对照组明显低于模型组(P<0.05)。③右侧黑质超氧化物歧化酶活力:各用药预防组和各用药治疗组及对照组明显高于模型组(P<0.05)。结论:银杏叶提取物及银杏总内酯可以预防和对抗6-羟基多巴造成的黑质多巴胺能神经元损伤。

AIM： To study the effects of ginkgo biloba extract （GBE） and bilobalide on cellular injury of substantia nigra of model rats with Parkinson disease （PD）. METHODS： The experiment was completed at the Animal Laboratory of Jinzhou Medical College in July 2004. Totally 80 healthy adult male SD rats were selected in this study. ① Prophylactic medication： Forty rats were randomly divided into 5 groups with 8 in each group. GBE prevention group： Rats were perfused with 100 mg/kg GBE solution including 24% chromocor, 6% bilobalide （provided by the Fourth Pharmaceutical Factory of Beijing, batch number. 0310469, 5 mL/branch）. Prevention group of high dosage of bilobalide： Rats were perfused with 12 mg/kg bilobalide solution （provided by Medical Institute of Capital University of Medical Sciences, batch number. 2003196, 2 mg/branch）. Prevention group of low dosage of bilobalide： Rats were perfused with 6 mg/kg bilobalide solution. Rats in control group and model group were perfused with the same volume of saline. All rats were medicated once a day for 14 successive days. One hour after the last medication, PD rats in modal and medication groups were modeled with 6-hydroxydopamine （6-OHDA） cerebral stereotactic injection. ② Remedial medication： Among other 40 rats, 8 rats were regarded as control group and perfused with the same volume of saline. Other 32 PD rats were modeled successfully and randomly divided into 4 groups with 8 in each group. GBE prevention group： Rats were perfused with 100 mg/kg GBE solution. Prevention group of high dosage of bilobalide： Rats were perfused with 12 mg/kg bilobalide solution. Prevention group of low dosage of bilobalide： Rats were perfused with 6 mg/kg bilobalide solution. Rats in model group were perfused with the same volume of saline. All rats were medicated once a day for 14 successive days. ③ All rats in prophylactic medication group after modeling and in remedial medication group after 14-day medication were killed to obtain brain and separate corpus striatum on beth sides. Concentration of dopamine was measeured with high pressure liquid chromatograph ultraviolet detector. Contents of superoxide dismutase （SOD） and malondialdehyde （MDA） in right substantia nigra were measured according to instruction provided by Nanjing Jiancheng Biological Institute. ④ Measurement data were compared with t test. RESUILTS： All 80 rats entered the final analysis.① Content of dopamine： Numbers of corpus striatum in injured side of rats were higher in prevention groups of GBE, high and low dosage of bilobalide than those in model group （P 〈 0.05-0.01）, but numbers in healthy side were not significant difference in each group （P 〉 0.05）. Numbers of corpus striatum in injured side of PD rats were higher in treatment group of high dosage of bilobalide than those in model group （P 〈 0.05）, and numbers in control group were higher than those in model group （P 〈 0.01）, but numbers in healthy side were not significant difference in each group （P 〉 0.05）. ② Content of MDA in right substantia nigra：Contents in prophylactic medication group, remedial medication group and control group were lower than those in model group （P 〈 0.05）.③ Activity of SOD in right substantia nigra： Activity in prophylactic medication group, remedial medication group and control group was higher than those in model group （P 〈 0.05）. CONCLUSION： GBE and bilobalide can protect doparnine neurons of substantia nigra from the damage of 6-OHDA.