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研究灵芝多糖对β淀粉样蛋白1-40(Aβ1-40)诱导体外培养大鼠海马神经元凋亡的保护作用以及可能的作用机制。体外培养新生Wistar大鼠海马神经元,MTT法检测不同浓度灵芝多糖干预Aβ1-40的细胞毒作用,选择出现显著拮抗细胞毒性的剂量为使用剂量。用HO33342检测细胞凋亡情况,用RT-PCR技术检测凋亡相关基因bcl-2、bax和caspase3基因mRNA的表达水平。结果显示10μmol/L Aβ1-40与海马神经元共孵育48h有明显的细胞毒作用,并可诱导神经元凋亡,加入不同浓度灵芝多糖后凋亡细胞明显不同程度减少。Aβ1-40可使bcl-2基因表达下调,bax基因表达上调,caspase-3基因的表达增强。加入灵芝多糖干预后,部分拮抗了Aβ1-40诱导的bcl-2基因的表达变化,从而使caspase-3基因的表达降低。灵芝多糖可通过调控bcl-2/bax的比值,降低caspase-3基因mRNA的表达来阻断Aβ1-40所诱导的海马神经元凋亡,具有神经保护作用。
To study the protective effect of Ganoderma lucidum polysaccharides on neuronal apoptosis induced by β-amyloid 1-40 (Aβ1-40) in vitro and its possible mechanism. The hippocampal neurons of newborn Wistar rats were cultured in vitro. The cytotoxic effect of Ganoderma lucidum polysaccharides with different concentration of Ganoderma lucidum on Aβ1-40 was detected by MTT method. The doses that were significantly antagonistic to cytotoxicity were selected. Apoptosis was detected by HO33342, and the mRNA expression of apoptosis-related genes bcl-2, bax and caspase3 were detected by RT-PCR. The results showed that 10μmol / L Aβ1-40 co-incubated with hippocampal neurons 48h had obvious cytotoxicity and induced apoptosis of neurons. After addition of different concentrations of Ganoderma lucidum polysaccharides, apoptotic cells decreased obviously. Aβ1-40 can down-regulate bcl-2 gene expression, up-regulate bax gene expression, and increase caspase-3 gene expression. The intervention of Ganoderma lucidum polysaccharide partially antagonized the change of bcl-2 gene expression induced by Aβ1-40, and thus reduced the expression of caspase-3 gene. Ganoderma lucidum polysaccharides can block the Aβ1-40-induced apoptosis of hippocampal neurons by regulating the ratio of bcl-2 / bax and reducing the expression of caspase-3 mRNA, which has the neuroprotective effect.