西洋参皂苷对缺氧性神经细胞坏死和凋亡的影响

Influence of Panax quinquefolium L. saponin on necrosis and apoptosis of cerebral cortical neurons induced by hypoxia

目的:探讨西洋参皂苷(PQLS)对缺氧诱导的胎鼠大脑皮层神经细胞抗凋亡的保护作用。方法:实验分为正常对照组、凋亡阳性组、西洋参皂苷0.025g/L组、0.05g/L组、0.1g/L组、0.2g/L组。采用无血清体外培养SD胚大鼠大脑皮层神经细胞;建立缺氧-复氧性凋亡细胞模型;Trypan blue拒染法、MTT法、Hoechst33342荧光染色法、Annexin V-FITC/PI流式细胞术等观察西洋参皂苷的毒性剂量及对缺氧性神经细胞坏死和凋亡的保护作用。结果:①PQLS在0.05～0.1g/L范围内促进神经细胞增殖较正常对照组提高9.78%～18.89%(P﹤0.01);在缺氧条件下PQLS在0.05～0.2g/L范围内促进神经细胞增殖较凋亡阳性组提高25.62%～37.65%(P﹤0.05～0.01),而在0.05%～0.1g/L(87.09%～91.39%)范围内与正常对照组(100.00%)差异无显著性(P 0.05)。②Annexin V-FITC/PI染色流式细胞仪测定表明,PQLS在0.025g/L～0.2g/L凋亡率(6.13±0.80%～14.47±0.60%)均显著低于凋亡阳性组(28.77±0.61%,P<0.01),但高于正常对照组(2.05±0.17%,P<0.01);而PQLS在0.025g/L～0.2g/L坏死率(2.15±0.17%～5.47±0.21%)均显著低于凋亡阳性组(6.57±0.23%,P 0.01),但高于正常对照组(0.84±0.09%,P<0.01)。③Hoechst 33342荧光染色证明,PQLS在0.025g/L～0.2g/L凋亡率(16.07±0.71%～29.91±1.06%)均显著低于凋亡阳性组(39.47±0.92%,P<0.01),而高于正常对照组(8.51±0.68%,P<0.01)。结论:西洋参皂苷具有促进神经细胞生长,阻止缺氧引起的神经细胞生长抑制、以及坏死和凋亡。

Objective：To explore the effects of Panax quinquefolium L.saponin（PQLS） on necrosis and apoptosis of embryonic rat cerebral cortical neurons induced by anoxia.Methods：The Experiments were divided into normal control,apoptosis group,PQLS 0.025g/L group,PQLS 0.05g/L group,PQLS 0.1g/L group,PQLS 0.2g/L group.Primary culture of embryonic rat cerebral cortical neurons was performed in serum-free in vitro.A model of apoptosis of cerebral cortical neurons was established by hypoxia for 12h then reoxygenation for 48h.The cytotoxic dose and protection of anoxic neuron of PQLS were determined by trypan blue exclusion staining,MTT assay,Hoechst33342 fluorescent staining and Annexin V-FITC/PI Flow cytometry.Results：①The PQLS within 0.05g/L ~ 0.1g/L promoting embryonic rat cerebral cortical neurons proliferation increased 9.78%~18.89% comparing with normal control in normoxia（P﹤0.01）.However,the PQLS within 0.05g/L~ 0.2g/L promoting embryonic rat cerebral cortical neurons proliferation increased 25.62% ~ 37.65% comparing with apoptosis group（P﹤0.05 0.01） and within 0.05g/L ~ 0.1g/L（87.09%~91.39%） it was not statistical difference comparing with normal control（100.00%）（P 0.05） in anoxia.②The apoptosis rates in PQLS within 0.025g/L ~ 0.2g/L（6.13±0.80%~14.47±0.60%） were lower than in apoptosis group（28.77±0.61%）（P 0.01） but were higher than in normal control（2.05±0.17%）（P 0.01）,and the necrosis rates in PQLS within 0.025g/L ~0.2g/L（2.15±0.17%~5.47±0.21%） were lower than in apoptosis group（6.57±0.23%）（P 0.01） but were higher than in normal control（0.84±0.09%）（P 0.01） through the Annexin V-FITC/PI staining flow cytometry determination.③The apoptosis rates in PQLS within 0.025g/L ~ 0.2g/L（16.07±0.71%~29.91±1.06%） were lower than in apoptosis group（39.47±0.92%）（P 0.01） but were higher than in normal control（8.51± 0.68%）（P 0.01） through the Hoechst 33342 fluorescent staining.Conclusion：Panax quinquefolium L.saponin is able to promote growth of cultured embryonic rat cerebral cortical neurons,and prevent growth suppression,necrosis and apoptosis of cultured embryonic rat cerebral cortical neurons caused by anoxia.