松生拟层孔菌水提物在体外对多巴胺能神经元的保护作用研究

Study on Neuroprotective Effects of Water Extract of Fomitopsis Pinicola on Dopaminergic Neurons in vitro

目的观察松生拟层孔菌水提物在体外对1-甲基-4-苯基吡啶(1-Methyl-4-phenylpyridine,MPP+)诱导胎鼠中脑多巴胺能细胞元凋亡的保护作用。方法采用酪氨酸羟化酶(tyrosine hydroxylase,TH)染色标记胎鼠中脑多巴胺能神经元,观察MPP+诱导后神经元数量和轴突长度的变化,通过检测还原型烟酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide,NADH)下降率来观察细胞线粒体呼吸链复合酶活性的变化,采用碘化乙啶(propidiumiodide,PI)染色分析细胞凋亡率。结果 MPP+诱导后TH阳性(THir)神经元数量明显减少,轴突长度明显下降,线粒体呼吸链复合酶I活性明显下降,凋亡率明显升高;松生拟层孔菌水提物作用后能明显增加THir神经元数量和轴突长度,其中50、25μg·mL^(-1)剂量组与MPP+组比较具有显著性差异;其可明显升高线粒体呼吸链复合酶I活性,其中50、25、12.5μg·mL^(-1)剂量组与MPP+组比较具有显著性差异;其可明显降低细胞凋亡率,其中50、25μg·mL^(-1)剂量组与MPP+组比较具有显著性差异。结论松生拟层孔菌水提物对MPP+诱导的胎鼠中脑多巴胺能细胞元凋亡具有明显的保护作用。

Objective To observe the neuroprotective effects of water extract of Fomitopsis Pinicola on 1-Methyl-4-phenylpyridine （MPP＋） induced apoptosis of mesencephala dopaminergic cells in vitro. Methods Mesencephalic dopamin-ergic neurons were labeled by tyrosine hydroxylase（TH） staining to observe the survival of THir neurons and the average length of neurite growth. The activity of mitochondrial respiratory chain complex I was analyzed by descent rate of nicotinamide adenine dinucleotide （NADH）. The relative apoptosis rate was analyzed by PI staining. Results The survival of TH immuno-reactive（THir） neurons and the average length of neurite growth increased obviously after MPP＋ induction, the activity of mitochondrial respiratory chain complex I decreased remarkably, and the apoptosis rate increased significantly. Compared with the MPP＋ control group, the survival of THir neurons and the average length of neurite growth were increased by water extract of Fomitopsis Pinicola at doses of 50 and 25 μg·mL-1 respectively. Water extract of Fomitopsis Pinicola treatment（50, 25 and 12.5 μg·mL^-1）dramatically increased the activity of mitochondrial respiratory chain complex I compared with MPP＋ control respectively. Compared with the MPP＋ control, the relative rates of PI-positive cells were significantly decreased by water extract of Fomitopsis Pinicola at doses of 50 and 25 μg·mL^-1 respectively. Conclusion Water extract of Fomitopsis Pinicola showed significant neuroprotective effect on mesencephala dopaminergic cells induced by MPP＋.