银杏内酯B对谷氨酸诱导脑皮质神经元凋亡及Ca^(2+)超载的影响  

Effects of Ginkgolide B on glutamate-induced apoptosis and calcium overload in cultured rat cortical neurons

作　　者：蔡卫斌[1] 杨中汉[1] 李朝阳[2] 宋志宏[1] 周世豪[1] 骆晓枫[1] 高国全[1] 

机构地区：[1]中山大学中山医学院生物化学教研室,广东广州510089 [2]中山大学中山眼科中心,广东广州510089

出　　处：《中国病理生理杂志》2005年第4期652-656,共5页Chinese Journal of Pathophysiology

基　　金：中华医学基金会资助项目 (CMB -SUMS学者项目 # 98- 6 77) ;教育部回国人员基金资助项目 (2 0 0 3年 ) ;广东省医学科研基金资助项目 (No .A2 0 0 315 3)

摘　　要：目的:观察银杏内酯B对谷氨酸诱导培养脑皮质神经元凋亡的拮抗作用,并探讨这种作用与神经细胞内游离Ca2 + 浓度改变的关系。方法:采用改良的方法原代培养胎小鼠脑皮质神经元,用噻唑兰(MTT)法检测神经元的存活情况;细胞凋亡采用形态学观察、DNA琼脂糖凝胶电泳法和Hoechst332. 5 8核染色方法进行分析;用Fura - 2 /AM荧光指示剂法测定细胞内Ca2 + 浓度。结果:谷氨酸(0 .8mmol/L)能诱导神经细胞凋亡和胞内Ca2 + 超载,银杏内酯B(10 - 2 5 0 μmol/L)能减轻谷氨酸所致的细胞损伤,表现为神经元存活率提高,细胞形态的恢复和DNA断裂减少。结论:银杏内酯B可拮抗谷氨酸所致的神经细胞毒性作用,这可能与其能竞争PAF受体并降低神经细胞内[Ca2 + ]从而抑制谷氨酸诱导的神经元凋亡有关。AIM: To observe the effects of ginkgolide B on the neuron apoptosis induced by glutamate and explore whether this effects a re related to the changes of calcium in neurons. METHODS: Primary neuron culture was prepared according to a prev iously reported procedure with slight modification. Neuron damage was induced by 0.8 mmol/L glutamate. The cell survival rate was examined by 3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays. The neuron morpholo gical changes, Hoechst 33258 unclear-staining analysis and agarose gel electroph oresis of DNA were measured as the indexes of cell apoptosis. Intracellular free Ca 2+ concentration in neurons was measured by using the fluorescent Ca 2+ indicator Fura-2/AM. RESULTS: The cells, exposed to glutamate (0.8 mmol/L), showed c haracteristic change of apoptosis and calcium overload, which were relieved by t he treatment of ginkgolide B (10-250 μmol/L), with survival increasing and cell morphology restoring and DNA fragment decreasing. CONCLUSIONS: Ginkgolide B prevents the neurons from glutamate ne urotoxicity by inhibiting glutamate-induced apoptosis. The potential mechanism o f its action may be related to the competitive PAF receptor binding of ginkgolid e B and decreasing the intracellular calcium concentration in neurons.

关 键 词：银杏内酯B 谷氨酸 脑皮质 神经元凋亡 CA^2+超载 

分 类 号：R741[医药卫生—神经病学与精神病学] R971[医药卫生—临床医学]