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目的 观察银杏叶提取物 (EGb)对复制的在体及离体帕金森病 (PD)模型的保护作用 ,并探讨其作用机制。方法 1 Wista大鼠 4 0只 ,雌雄不拘 ,体重 2 30± 2 0 g ,随机分成 4组 (各组n =10 )。制模前 19d ,第Ⅰ、Ⅱ组腹腔内注射等量生理盐水 ,第Ⅲ、Ⅳ组分别腹腔内注射EGb 50mg/kg、10 0mg/kg ,每天 1次。第 2 0天制模 :第Ⅰ组左侧黑质内注射等量人工脑脊液 (ACSF) ,第Ⅱ、Ⅲ、Ⅳ组左侧黑质内注射 1 甲基 4 苯基 1,2 ,3,6 四氢吡啶 (MPTP) 40 μg/kg(2 μg/ μl,左侧黑质坐标为 :AP 5 4mm ,L 2 .2mm ,H 8.3mm )。制模后 12h ,第Ⅲ、Ⅳ组再分别腹腔内注射EGb 50mg/kg、10 0mg/kg 1次。制膜后 2 4h断头处死 ,取损毁侧黑质、纹状体充分匀浆 ,分别测定其中丙二醛 (MDA)、超氧化物歧化酶 (SOD)、多巴胺 (DA)的含量变化。 2 ① 1 甲基 4 苯基 吡啶离子 (MPP+ )诱导PC12细胞凋亡 :将PC12细胞种植在内有盖玻片的 6孔培养板中。 4 8h后 3、4、5、6孔内均加入MPP+ (终浓度为 10 μM ) ,1、2孔作对照。 6h后取出细胞爬片 ,常规固定 ,H E染色 ,光镜下观察凋亡细胞 ;免疫组化检测P53、bcl 2蛋白的表达情况。同样实验重复 3次。②观察EGb对MPP+ 诱导PC12细胞凋亡的影响 :PC12细胞在 6孔板中 (3块 )生长 4 8h后 ,
Objective To observe the protective effect of Ginkgo biloba extract (EGb) on replicating in vitro and in vivo Parkinson’s disease (PD) models and to explore its mechanism of action. Methods 1 40 Wistar rats, male or female, weighing 2 30±20 g, were randomly divided into 4 groups (n=10 for each group). On the 19th day before the injection of mold, groups I and II received intraperitoneal injection of normal saline. Groups III and IV were injected intraperitoneally with EGb 50 mg/kg and 100 mg/kg once daily. Day 20: The left intraperitoneal injection of equal amount of artificial cerebrospinal fluid (ACSF) in group I and the left substantia nigra injection of 1 methyl 4-phenyl 1,2,3,6 in groups II, III, and IV Tetrahydropyridine (MPTP) 40 μg/kg (2 μg/μl, left subsurface coordinates: AP 5 4 mm, L 2 .2 mm, H 8.3 mm ). After 12 hours of molding, groups III and IV were injected intraperitoneally with EGb 50 mg/kg and 100 mg/kg, respectively. After 24 hours of membrane production, the rats were sacrificed by decapitation, and the substantia nigra and striatum were homogenized. The contents of malondialdehyde (MDA), superoxide dismutase (SOD), and dopamine (DA) were measured. 2 1 1 Methyl 4-phenylpyridinium (MPP+) induces apoptosis in PC12 cells: PC12 cells were seeded in 6-well culture plates with coverslips. After 4 hours, 3, 4, 5, and 6 wells were added with MPP+ (final concentration 10 μM). One and two wells were used as controls. After 6 hours, the cells were removed and routinely fixed and HE stained. Apoptotic cells were observed under light microscope. The expression of P53 and bcl-2 protein was detected by immunohistochemistry. The same experiment was repeated 3 times. 2 Observe the effect of EGb on the apoptosis of PC12 cells induced by MPP+:PC12 cells grow in the 6-well plate (3 cells) for 48 hours.