白扁豆多糖对神经细胞缺氧性凋亡的保护  

Protective effect of white hyacinth bean polysaccharide on the apoptosis of fetal rat cerebral cortical neurons induced by hypoxia

作　　者：胡国柱[1] 姚于飞 文珠[3] 高幼奇[1] 

机构地区：[1]江西省人民医院,南昌330006 [2]乐清市人民医院ICU科,乐清325600 [3]江西省医学科学研究所,南昌330006

出　　处：《中药药理与临床》2012年第1期91-94,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：江西省科技厅2006年社会发展项目

摘　　要：目的:探讨白扁豆多糖(WHBP,white hyacinth bean polysaccharide)抗神经细胞缺氧性凋亡的作用机制。方法:实验分正常对照组,凋亡诱导组,WHBP 0.5g/L组、3.5g/L组、8.0g/L组;采用Neurobasal加B27 Supplement体外培养SD胚大鼠大脑皮层神经细胞;应用MTT法、Trypan blue拒染法、DNA琼脂糖电泳法、免疫细胞化学染色等观察细胞活性和凋亡。结果:①MTT测定证明缺氧条件下WHBP 2.5g/L～3.5g/L刺激神经细胞的活性与正常对照组比较无差异(P>0.05),而与凋亡诱导组比较WHBP 0.5g/L～6.0g/L刺激神经细胞的活性均显著增高(P<0.000)。②DNA琼脂糖凝胶电泳发现WHBP 3.5g/L组与正常对照组相似未见有明显凋亡梯状条带。③免疫细胞化学发现缺氧条件下WHBP 3.5g/L刺激神经细胞的Bcl-2表达率(63.01±1.40%)高于凋亡诱导组(37.83±7.41%),且与正常对照组相近(68.07±3.17%);Bax和Caspase-3表达率(62.79±1.30%,45.75±3.44%)显著低于凋亡诱导组(72.09±3.98%,70.01±3.43%)(P<0.05～0.000),且与正常对照组相比情况不一(63.36±2.22%,33.60±4.14%)(P>0.05,P<0.01);但是Bcl-2/Bax比值(1.01±0.02)显著高于凋亡诱导组(0.52±0.08)(P<0.000),而与正常对照组相近(1.07±0.08)(P>0.05)。结论:WHBP通过减少Bax、Caspase-3的表达,相对提高Bcl-2的表达及Bcl-2/Bax比例阻断缺氧诱导的神经细胞凋亡和保护神经细胞。Objective ：To investigate the mechanism of white hyacinth bean polysaccharide（WHBP） anti-apoptosis of fetal rat cerebral cortical neurons induced by hypoxia.Methods ：The experiments were divided into normal control,apoptosis group,WHBP 0.5g/L group,WHBP 3.5g/L group,WHBP 8.0g/L group.To culture fetal rat cerebral cortical neurons in serum-free neurobasal medium supplied with 2%B27 supplement.Using MTT,trypan blue exclusion staining,DNA agarose electrophoresis and immunocytochemical staining to observe cell activity and cell apoptosis.Results： The cell activity was not different between WHBP 2.5g/L^3.5g/L stimulating hypoxia-induced fetal rat cerebral cortical neurons and neurons in normal control（P0.05）,but the cell activity significantly increased in WHBP 0.5g/L^6.0g/L stimulating hypoxia-induced fetal rat cerebral cortical neurons comparing with ones in apoptosis group（P0.000）.In WHBP 3.5g/L group the apoptotic ladder of DNA fragments extracted from hypoxia-induced neurons was not obviouslike one innormal control.In WHBP 3.5g/L stimulating hypoxia-induced neurons group the Bcl-2 expression（63.01±1.40%） was higher than in apoptosis group（37.83±7.41%） and very similar（68.07±3.17%） with normal group.In WHBP 3.5g/L stimulating hypoxia-induced neurons group Bax and Caspase-3 expressions（62.79±1.30%,45.75±3.44%） were significantly lower than in apoptosis group（72.09±3.98%,70.01±3.43%）（P0.05~0.000） and similar（63.36±2.22%,P0.05） or higher（33.60±4.14%,P0.01） than in normal control.However,in WHBP 3.5g/L stimulating hypoxia-induced neurons group Bcl-2/Bax ratio（1.01±0.02） was significantly higher than in apoptosis group（0.52±0.08,P0.000） and very similar（1.07±0.08,P0.05） with normal control.Conclusions： The effect of WHBP protecting hypoxia-induced neurons from apoptosis is by reducing Bax and Caspase-3 expressions and the relative increase of Bcl-2 expression and Bcl-2/Bax ratio.

关 键 词：白扁豆多糖 神经细胞 原代培养 缺氧 凋亡 BCL-2 BAX CASPASE-3 

分 类 号：R285[医药卫生—中药学]