白扁豆多糖抗神经细胞缺氧性坏死与凋亡

The effect of white hyacinth bean polysaccharide on the anti-apoptosis and necrosis of fetal rat cerebral cortical neurons induced by hypoxia

目的:探讨白扁豆多糖(WHBP,white hyacinth bean polysaccharide)抗胚鼠大脑皮层神经细胞缺氧性坏死及凋亡的作用。方法:实验分正常对照组,凋亡阳性组,白扁豆多糖0.5g/L、3.5g/L、8.0g/L组;采用无血清体外培养SD胚大鼠大脑皮层神经细胞;应用MTT法、Trypan blue拒染法、Annexin V-FITC/PI流式细胞术等方法观察白扁豆多糖的合适使用剂量和缺氧性神经细胞坏死及凋亡保护作用。结果:①常氧条件下白扁豆多糖2.0g/L～6.0g/L促进神经细胞相对增殖率达28.85%～15.38%,与正常对照组比较P<0.01;而缺氧条件下白扁豆多糖2.5g/L～3.5g/L与正常对照组比较P>0.05,但与凋亡阳性组比较白扁豆多糖0.5g/L～6.0g/L相对增殖率均高(P<0.01)。②Annexin V-FITC/PI染色流式细胞测定表明,白扁豆多糖0.5g/L、3.5g/L、8.0g/L组的坏死率分别为9.45±5.28%、8.39±3.44%、9.85±2.90%,与凋亡阳性组(10.01±2.94%)及正常对照组(5.38±1.46%)比较P>0.05;而早期凋亡率分别为19.83±2.50%(P<0.05)、9.34±2.47%(P<0.01)、23.35±5.14%,与凋亡阳性组(28.72±2.74%)比较差异显著,而与正常对照组(4.54±0.34%)比较除3.5g/L组外,其余两组均P<0.01。结论:白扁豆多糖具有促进神经细胞生长,阻断缺氧引起的神经细胞生长抑制,以及显著地抗神经细胞缺氧性凋亡功效。

To investigate the effect of white hyacinth bean polysaccharide（WHBP） on the anti-apoptosis and necrosis of fetal rat cerebral cortical neurons induced by hypoxia. Methods：To culture fetal rat cerebral cortical neurons in serum-free neurobasal medium supplied with 2% Bz/ supplement. The Experiments were divided into normal control, apoptosis positive group, WHBP 0.5g/L group, WHBP 3.5g/L group, WHBP 8.0g/L group. To observe optimal dose and protective effect of WHBP for hypoxic neuronal necrosis and apoptosis by using MTI＇, Trypan blue exclusion Staining, Annexin V-FITC/PI Flow cytometry and DNA agarose electrophoresis. Results： The relative prolifera- ting rate of cultured fetal rat cerebral cortical neurons stimulated by WHBP 2.0g/L - 6.0g/L was 28.85% - 15.38% in normoxia, compa- ring with normal control（ P 〈0.05）. The relative proliferating rates of cultured fetal rat cerebral cortical neurons stimulated by WHBP 2.5g/ L - 3.5g/L were higher than apoptosis positive group in hypoxia, and were not statistical difference in WHBP 2.5g/L-3.5g/L groups com- paring with in normal control （ P 〉 0.05 ）. The necrosis rates were not statistical difference in WHBP O. 5g/L（ 9.45±5.28 % ）, 3.5g/L（8.39±3.44% ）,8.0g/L（ 9.85± 2.90% ） groups comparing in apoptosis pisitive group （10.01±2.94% ） and in normal control （5.38±1.46% ） （ P 〉 0.05 ）. However, the early apoptosis rates were statistical difference in WHBP 0.5g/L（ 19.83±2.50%, P 〈 0.05 ） ,3.5g/L（9. 34±2.47% , P 〈 0.001 ）, 8.0g/L（23.35±5.14% ） groups comparing in apoptosis positive group（ 28.72±2.74% ） and in normal control （d. 54±0.34% ） （ P 〈0.001 ） except in WHBP 3.5g/L. Condusion：WHBP is able to promote growth of cultured fetal rat cerebral cortical neurons ,prevent growth suppression and apoptosis of cultured fetal rat cerebral cortical neurons caused by hypoxia.