黄精多糖对新生大鼠大脑皮层神经细胞缺氧性凋亡的影响

Influence of Polygonatum Polysaccharide on apoptosis of primary cultured neonate rat cerebral cortical neurons caused by hypoxia

目的探讨黄精多糖对体外培养的新生大鼠大脑皮层神经细胞缺氧性凋亡的保护作用。方法采用“Neurobasal加B27Supplement”体外培养新生大鼠大脑皮层神经细胞,使用Hoechst33342荧光染色、免疫细胞化学染色观察黄精多糖对缺氧复氧性神经细胞凋亡的保护作用。结果缺氧前加入500μg/ml^1.5mg/ml的黄精多糖能显著地降低缺氧复氧培养诱导的神经细胞凋亡率,增加缺氧的神经细胞Bcl-2蛋白的表达,减少Bax蛋白的表达,提高Bcl-2/Bax比值。而缺氧12h后加入黄精多糖则无明显的抗凋亡作用。结论缺氧前加入黄精多糖可以通过上调缺氧神经细胞Bcl-2表达、下调Bax表达和提高Bcl-2/Bax的比值以避免缺氧的神经细胞凋亡。

Objective： To investigate the protective mechanism of Pdygonatum Polysaccharide （PP） on apoptosis of primary cultured cerebral cortical neurons （PCCCN） caused by hypoxia. Methods： The anti-apoptosis effect of PP on primary cultured neurons was observed by means of the PCCCN of postnatal rats in free-serum with neurobasal medium supplied with 2%B27 supplement, hypoxie culture of neurons, Hoechst 33342 staining and immunocytoehemistry. Results： The PP added to neurons culture in 0.5, 1 and 1.5mg/ml before hypoxia markedly dropped apoptosis rate of neurons induced by hypoxia （P〈0.01）, increased Bcl-2 protein expression （P〈 0.01 ） and decreased Bax protein expression （P〈 0.01 ） in neurons induced by hypoxia, and raised the ratio of Bcl-2/Bax （P〈 0.01 ）. The PP added to neurons culture in 0. 5, 1 and 1.5mg/ml after hypoxia 12h hardly had anti-apoptosis effect （P〉0.05）. Conclusion： The PP added to neurons culture in 0.5, 1 and 1.5mg/ml before hypoxia is able to prevent from apoptosis of hypoxic neurons in PCCCN. The effect of anti-apoptosis is through upregulation of Bcl-2 protein expression , down-regulation of Bax-2 protein expression and raising the ratio of Bcl-2/Bax.