灵孢多糖对MPP^+损伤PC12细胞的保护作用

Ganoderma lucidum Polysaccharides Protects PC12 Cells against MPP^+ Induced Neurotoxicity

【目的】观察灵孢多糖(GLPS)对甲基-苯基-吡啶离子(MPP+)诱导损伤的PC12细胞的保护作用。【方法】将MPP+或GLPS加入体外培养的PC12细胞中,建立多巴胺能神经元损伤模型,用四甲基偶氮唑盐(MTT)检测细胞活力的变化;以乳酸脱氢酶(LDH)检测法检测培养上清液中LDH水平的改变;通过酪氨酸羟化酶(TH)免疫细胞化学法检测TH阳性细胞数及蛋白的表达。【结果】MPP+处理48h后,细胞活力降至对照组的52%,经GLPS(100、200、300!g/mL)预处理后,细胞活力明显提高,分别为65%,75%,79%(P<0.05)。MPP+处理48h后,上清液中LDH水平较对照组明显提高,经不同浓度的GLPS预处理后,上清中LDH水平有所下降(P<0.05),且TH阳性细胞数及表达强度明显高于MPP+组。【结论】灵孢多糖对MPP+诱导损伤的PC12细胞具有保护作用。

[Objective] To examine the neuroprotective effects of Ganoderma lucidum polysaccharides （GLPS） on 1-methyl-4-phenylpyridinium ion （MPP^＋） induced neurotoxicity in PC12 cells. [ Method ] Neurotoxicity was induced by addition of MPP^＋ into cultures of PC12 cells and different doses of GLPS were administrated 30 rain before addition of MPP^＋ into PC12 cell cultures. Cell viability and protein expression were measured using 3-（4,5- dimethyhhiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT）, lactate dehydrogenase （LDH） and immunohistochemical activities of tyrosine hydroxylase （TH）. [ Results ] The percentage of viability of PC12 cells treated with MPP^＋ for 48 h was 52% of control group in MTT assay. In addition, treatment with MPP^＋ increased release of LDH into medium and reduced immunohistochemical activities of TH neuron. Pretreatment with single dose of GLPS at 100, 200, and 300μg/mL significantly increased cell viability by 65%, 75%, and 79% in MTT assay respectively. In parallel, treatment with GLPS significantly reduced MPP^＋ induced LDH release and attenuated MPP^＋ induced reduction of TH immunohistochemical activities. [ Conclusion ] The results suggest that pretreatment of GLPS protects PC12 cells against MPP^＋ induced neurotoxicity.