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目的:研究灵芝发酵液诱导PC12细胞分化及对β淀粉样肽细胞毒性的保护作用。方法:制备灵芝发酵液,体外培养未分化PC12细胞,观察其在发酵产物的诱导下突起生长情况并计算分化率。细胞以发酵液预处理48 h后,加入终浓度20μmol/L的“老化”Aβ25-35肽,24 h后测定细胞MTT、ROS及培养液上清LDH、MDA。以倒置相差显微镜进行形态学观察并拍摄相片记录。结果:灵芝发酵液25、50、100 mg/L可明显提高PC12细胞分化率,对抗Aβ25-35肽导致的细胞存活率降低并降低细胞ROS水平;50、100mg/L能减少细胞LDH释放量及MDA含量。结论:灵芝发酵液能明显诱导PC12细胞分化,对Aβ25-35肽导致的神经毒性有良好的保护作用,提示其作用机制可能与促进神经发生及对抗神经细胞氧化损伤有关。
Objective: To study the protective effect of Ganoderma lucidum fermentation broth on the differentiation of PC12 cells and its cytotoxicity against β-amyloid peptides. Methods: The lucid ganoderma fermentation broth was prepared and the undifferentiated PC12 cells were cultured in vitro. The neurite outgrowth under the induction of the fermentation product was observed and the differentiation rate was calculated. The cells were pretreated with the fermentation broth for 48 h, and then added with an “ablated” Aβ25-35 peptide with a final concentration of 20 μmol/L. After 24 h, the cells were assayed for MTT, ROS, and supernatants of the culture medium for LDH and MDA. Morphological observations were performed with an inverted phase contrast microscope and photographs were taken. Results: Ganoderma lucidum fermentation broth 25, 50, 100 mg / L can significantly increase the differentiation rate of PC12 cells, against the Aβ25-35 peptide caused by decreased cell viability and decreased cellular ROS levels; 50,100 mg / L can reduce cell LDH release and MDA content. Conclusion: Ganoderma lucidum fermentation broth can obviously induce the differentiation of PC12 cells and has a good protective effect on the neurotoxicity induced by Aβ25-35 peptide. It suggests that the mechanism of action may be related to the promotion of neurogenesis and the protection against oxidative damage of nerve cells.