人参皂甙Rb1抗新生大鼠大脑皮层神经细胞缺氧性凋亡研究

Study of the influence of Ginsenoside Rb1 on apoptosis of primary cultured neonate rat cerebral cortical neurons caused by hypoxia

目的探讨人参皂甙Rb1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机理。方法应用“Neurobasal加B2 7Supplement”体外培养大鼠大脑皮层神经细胞 ,并在缺氧条件下使用台盼蓝拒染法、Hoechst 3 3 3 42荧光染色法、免疫细胞化学染色法观察人参皂甙Rb1对原代神经细胞的抗凋亡保护作用。结果在 10— 10 0 μg/ml的浓度范围内 ,人参皂甙Rb1能降低缺氧诱导的神经细胞凋亡率 (10 0 μg/ml组 ,P <0 .0 5 ) ,增加缺氧神经细胞Bcl 2蛋白的表达 (5 0— 10 0 μg/ml组 ,P <0 .0 5 ) ,同时减少Bax蛋白的表达 (5 0— 10 0 μg/ml组 ,P <0 .0 5—P <0 .0 0 1) ,提高Bcl 2 /Bax比值 (5 0— 10 0 μg/ml组 ,P <0 .0 5 )。 结论在 5 0— 10 0μg/ml的浓度范围内 ,人参皂甙Rb1通过上调缺氧神经细胞Bcl 2表达和下调Bax表达 ,避免缺氧神经细胞凋亡。

ObjectiveTo investigate the protective mechanism of Ginsenoside Rb1 on apoptosis of primary cultured cerebral cortical neurons caused by hypoxia.MethodsThe anti apoptosis effect of Ginsenoside Rb1 on primary cultured neurons was observed by methods of the primary culture of cerebral neurons of postnatal rats in free serum with neurobasal medium supplied with 2% B27 supplement, trypan blue exclusion, hypoxic culture of neurons, Hoechst 33342 staining and immunocytochemistry.ResultsAt concentrations of 10 μg/ml, 50 μg/ml and 100 μg/ml, the Ginsenoside Rb1 dropped apoptosis rate of cerebral cortical neurons induced by hypoxia (in 100 μg/ml, P<0.05), and increased Bcl 2 protein expression (except 10 μg/ml, P<0.05) and decreased Bax protein expression (except 10 μg/ml, P<0.05— P<0.001) in the cerebral cortical neurons induced by hypoxia, improved the ratio of Bcl 2/Bax (except 10 μg/ml, P<0.05).ConclusionGinsenoside Rb1 is able to prevent hypoxic neurons from apoptosis in primary cultured cerebral cortical neurons from 50—100 μg/ml. The effect of anti apoptosis is through up regulation of Bcl 2 protein expression and down regulation of Bax 2 protein expression. [