白果内酯对糖氧剥夺小鼠原代海马神经元内钙离子浓度的影响  

Research on the influence and mechanism of bilobalide on intracellular calcium concentration of primary hippocampal neurons in mice

作　　者：王蕾 张雄 陈赵乐 王玉同 Wang Lei;Zhang Xiong;Chen Zhaole;Wang Yutong(Department of Emergency of Xijing Hospital,Air Force Medical University,Xi’an 710032,China)

机构地区：[1]空军军医大学西京医院急诊科,西安710032

出　　处：《神经解剖学杂志》2020年第1期40-44,共5页Chinese Journal of Neuroanatomy

摘　　要：目的:观察白果内酯对糖氧剥夺(OGD)后小鼠原代海马神经元内游离钙离子浓度的影响及机制探究。方法:分离和培养小鼠原代海马神经元,实验分为对照组(control)、糖氧剥夺组(OGD)和白果内酯组(OGD+bilobalide),对照组予以常规培养小鼠原代海马神经元,糖氧剥夺组利用氧葡萄糖剥夺/再恢复模型模拟小鼠原代海马神经元的缺血再灌注损伤,白果内酯组在氧葡萄糖剥夺后再恢复前应用20μmol/L的白果内酯。利用流式细胞仪检测各组小鼠原代海马神经元内游离的钙离子浓度,利用Western Blot技术检测各组小鼠原代海马神经元Cav1.2蛋白的表达,最后利用膜片钳技术检测各组小鼠原代海马神经元L型电压依赖型钙离子通道的功能。结果:与糖氧剥夺组相比,白果内酯组小鼠原代海马神经元游离钙离子浓度降低(P<0.05)、Cav1.2蛋白表达量减少(P<0.05)、L型电压依赖性钙离子通道功能减弱(P<0.05),但仍未恢复到对照组水平(P<0.05)。结论:白果内酯可能通过减少小鼠原代海马神经元Cav1.2表达和L型电压依赖性钙离子通道的功能降低细胞内游离钙离子浓度,从而发挥对糖氧剥夺后神经元的保护作用。Objective:To investigate the influence and mechanism of bilobalide on intracellular calcium concentration of primary hippocampal neurons in mice.Methods:The mice primary hippocampal neurons were isolated and randomly divided into control group,oxygen-glucose deprivation group(OGD)and bilobalide group(OGD+bilobalide).The neurons in the control group were cultured in the conventional incubator,and the neurons in OGD group were cultured by oxygen-glucose deprivation/reperfusion(OGDR)method to simulate ischemia reperfusion injury.The neurons in the OGD+bilobalide group were treated with 20μmol/L bilobalide after incubation in hypoxic incubator and before incubation in conventional incubator.After exposure to 20μmol/L bilobalide,we tested the intracellular calcium concentration of mice primary hippocampal neurons by flow cytometry.The expression of Cav1.2 was measured by the Western Blot.The function of L-type calcium channel(LTCC)was measured by patch clamp.Results:Compared with the OGD group,the free calcium concentration,the expression of Cav1.2 protein and the function of LTCC in the primary hippocampal neurons of mice in the OGD+bilobalide group decreased(P<0.05),but they still did not return to the level of the control group(P<0.05).Conclusion:Bilobalide may decreases the intracellular calcium concentration of mice primary hippocampal neurons by inhibiting the expression of Cav1.2 and the function of LTCC.

关 键 词：白果内酯 糖氧剥夺 细胞内钙离子浓度 L型电压依赖性钙离子通道 Cav1.2 小鼠原代海马神经元 

分 类 号：R285.5[医药卫生—中药学]