低分子量姬松茸多糖抑制氧应激大鼠海马神经元的凋亡作用

Inhibition function of low molecular weight polysaccharide from Agaricus blazei on the apoptosis of hippocampal neurons in rats with oxygen stress

目的研究低分子量姬松茸多糖(LMPAB)对氧应激大鼠海马神经元Bcl-2和Bax表达的影响。方法取24 h内出生大鼠的海马神经元细胞原代培养,分为对照组、模型组、3个剂量实验组(LMPAB,3,5,10,20 mg·L^(-1))。实验组预处理24 h后,同模型组均加入500μmol·L^(-1) H_2O_2损伤2 h。用流式细胞术和MTT法检测大鼠海马神经元细胞凋亡及细胞活性,用免疫细胞化学检测Bcl-2和Bax蛋白的表达。结果海马神经元凋亡率,高剂量组(10.08±0.83)显著低于模型组(29.01±0.67,P<0.01);细胞活力,高剂量组(0.38±0.05)显著高于模型组(0.20±0.07,P<0.01);与模型组相比,高剂量组Bcl-2表达显著增高(42.73±3.65 vs 29.43±2.26),Bax表达显著降低(22.47±2.82 vs47.58±4.79)(均P<0.01),Bcl-2/Bax比率增高。结论 LMPAB对氧化应激损伤的大鼠海马神经元有一定的保护作用,其机制可能与抑制海马神经元凋亡有关。

Objective To investigate the effect of low molecular weight polysaccharide from Agaricus blazei（ LMPAB） on the expression of Bcl- 2 and Bax in rat primary hippocampal neurons（ PHN） by oxidative stress. Methods PHN were clipped from 24 h- old rat and cultured primarily. Different groups were set,including the control group,model group and test groups（ 5,10 and 20 mg·L^-1）. At 24 h after LMPAB pretreatment, oxidative damage of cells were induced with 500μmol·L^-1 H2O2 for 2 h in model and test group. The PHN apoptosis and cell activity were measured with flow cytometry and MTT method.The expression of Bcl- 2 and Bax protein were detected by immunocytochemical technique. Results Hippocampus neuron apoptosis rate in high dose test group（ 10. 08 ± 0. 83） was significantly lower than that of model group（ 29. 01 ± 0. 67,P〈0. 01）. Cell viability in high dose group（ 0. 38 ± 0. 05） was significantly higher than that of model group（ 0. 20 ± 0. 07,P〈0. 01）. Compared with model group, Bcl- 2 expression significantly increased（ 42. 73 ± 3. 65 vs 29. 43 ± 2. 26）.However, Bax expression significantly reduced in high dose group（ 22. 47 ± 2. 82 vs 47. 58 ± 4. 79）（ P〈0. 01）,Bcl- 2 /Bax increased.Conclusion LMPAB attenuated the hippcampal neurons oxidative damageinduced by H2O2. The mechanism may be related to its inhibition on the apoptosis of hippocampal neurons.