摘要
目的研究灵芝孢子对帕金森病(PD)模型鼠脑黑质酪氨酸羟化酶(TH)的影响,以探讨灵芝孢子对PD的脑保护作用。方法120只SD大鼠随机分为三组,PD组:立体定向注入6-羟多巴(6-OHDA),术后1周腹腔注入阿朴吗啡观察30min大鼠旋转的次数,连续至第四周每分钟大于6次者为成功的PD模型;灵芝孢子组:先用灵芝孢子粉灌胃3d,立体定向注入6-OHDA,继续灌胃4周,直至处死;正常对照组:立体定向注入脑黑质抗坏血酸生理盐水。处死后制作快速冰冻切片用免疫组化检测TH阳性细胞数,用原位杂交法检测THmRNA的阳性细胞数,用Westernblot检测TH的半定量变化,用高效液相色谱检测多巴胺水平的变化。结果免疫组化显示灵芝孢子组术侧鼠脑黑质TH阳性神经元数量为99·7±13·6,PD组为51·0±13·5,灵芝孢子组较PD组显著升高(P<0·05);原位杂交显示灵芝孢子组术侧鼠脑黑质THmRNA阳性神经元数量为180·3±24·3,PD组为72·7±15·0,灵芝孢子组较PD组显著升高(P<0·01);Westernblot检测显示TH蛋白灵芝孢子组较PD组显著增加;高效液相色谱检测术侧鼠脑黑质多巴胺(DA)水平灵芝孢子组为(0·7±0·3)ng/mg蛋白,PD组为(0·4±0·1)ng/mg蛋白,灵芝孢子组较PD组显著升高(P<0·05)。结论灵芝孢子能够提高TH的表达,并提高鼠脑黑质DA的水平,推测对6-OHDA诱导的PD可能有脑保护作用。
Objective To explore the neuroprotective effects of ganoderma spore on parkinsonian rat model. Methods 120 male Sprague-Dawley rats were divided into three groups: parkinsonian groups with and without treatment with ganoderma spore (daily for 4 weeks) and control group. The parkinsonian rat model was produced by unilaterally infusion with 6-hydroxydopamine (6-OHDA) in the right substantia nigra of rats. Apomorphine-induced rotation was counted as behavioral assessment. The expression levels of tyrosine hydroxylase (TH) protein and mRNA were measured by immunohistochemistry (IH), Western blot, and in situ hybridization (ISH). The number of TH-positive neurons and content of dopamine (DA) in the substantia nigra was determined. Results The number of TH positive cells in substantial nigra of rats treated with ganoderma spore was 99.7±13.6, similar to that of the control group (105.0±17.6) but significantly higher than that of untreated group (51.0±13.5, P<0.05). The expression of TH mRNA was also significantly increased in ganoderma spore treated rats as compared to the untreated PD rats. The level of DA in the ganoderma spore treated group (0.7±0.3 ng/mg protein) was higher than that of the untreated rats (0.4±0.1 ng/mg protein, P<0.05). Conclusion Our results suggest that ganoderma spore may provide a protective effect against 6OHDA-induced neuronal death in parkinsonian model.
出处
《中华神经科杂志》
CAS
CSCD
北大核心
2005年第6期355-358,共4页
Chinese Journal of Neurology
