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目的:观察银杏叶提取物(EGb)对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)及其离子1-甲基-4-苯基吡啶(MPP~+)诱导的帕金森症(PD)模型的保护作用。方法:用脑立体定位仪向黑质(AP-5.4mm,-2.2mm,H8.3mm)内注射MPTP诱导大鼠旋转。在注射MPTP 24h后将大鼠处死,硫代巴比妥法测定黑质中丙二醛(MDA),羟胺法(即改进的黄嘌呤氧化酶法)测定黑质中超氧化物歧化酶(SOD),荧光分光光度法(激发波长310nm,发射波长390nm)测定纹状体中多巴胺(DA)的含量。MPP~+诱导PC12细胞凋亡,HE染色,光镜下观察凋亡细胞;吖啶橙/溴乙锭(AO/EB)染色,荧光显微镜记数凋亡细胞,观察不同浓度EGb(25,50,100mg/L)在6h,12h,24h对细胞凋亡率的影响。结果:EGb 100mg/kg组可减少模型鼠的旋转次数及旋转持续时间(n=10,P<0.05);与MPTP组比较,EGb 50mg/kg和100mg/kg组MDA相对降低,SOD及DA相对增高(n=10,P<0.05和P<0.01)。MPP~+10μmol/L可诱导PC12细胞凋亡,EGb 50和100mg/L组在6h,12h,24h可降低细胞凋亡率(P<0.05和P<0.01,n=3)。结论:EGb对MPTP诱导的PD动物模型及其离子MPP~+诱导的PD细胞模型均有保护作用,其保护机制与清除自由基及抑制神经元凋亡有关。
Objective: To observe the effect of ginkgo biloba extract (EGb) on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its 1-methyl-4-phenylpyridine (MPP) ~+) induced Parkinson’s disease (PD) model protection. METHODS: MPTP was injected into the substantia nigra (AP-5.4mm, -2.2mm, H8.3mm) to induce rat rotation using a stereotaxic apparatus. Rats were sacrificed 24 hours after MPTP injection. Malondialdehyde (MDA) in substantia nigra was determined by thiobarbital, and superoxide dismutase (SOD) in substantia nigra was determined by hydroxylamine method (ie, modified xanthine oxidase method). Fluorescence spectrophotometry (excitation wavelength 310 nm, emission wavelength 390 nm) was used to determine the dopamine (DA) content in the striatum. MPP~+ induced apoptosis of PC12 cells, HE staining, apoptotic cells were observed under light microscope, acridine orange/ethidium bromide (AO/EB) staining, apoptotic cells were counted by fluorescence microscope, and different concentrations of EGb were observed (25,50,100). The effect of mg/L on apoptosis rate at 6h, 12h and 24h. Results: The EGb 100mg/kg group could reduce the number of rotation and duration of rotation in the model rats (n=10, P<0.05). Compared with the MPTP group, the EGb 50mg/kg and 100mg/kg group had relatively lower MDA, SOD and DA were relatively Increased (n=10, P<0.05 and P<0.01). MPP~+10μmol/L could induce apoptosis in PC12 cells, and EGb 50 and 100mg/L group could reduce the apoptosis rate at 6h, 12h and 24h (P<0.05 and P<0.01, n=3). CONCLUSION: EGb has protective effects on MPTP-induced PD animal model and its ion-MPP+ induced PD cell model. Its protective mechanism is related to scavenging free radicals and inhibiting neuronal apoptosis.