[1]严飞平　陈　晨　李志磊　霍国进　冯　毅　白　磊.茶多酚对氧葡萄糖剥夺/再恢复干预后原代培养的小鼠海马神经元内钙离子浓度的影响[J].中国临床神经外科杂志,2019,(10):608-610.[doi:10.13798/j.issn.1009-153X.2019.10.011]
　YAN Fei-ping,CHEN Chen,LI Zhi-lei,et al.Effect of tea polyphenol on intracellular calcium concentration of primary hippocampal neurons and its mechanism in mice[J].,2019,(10):608-610.[doi:10.13798/j.issn.1009-153X.2019.10.011]
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茶多酚对氧葡萄糖剥夺/再恢复干预后原代培养的小鼠海马神经元内钙离子浓度的影响()
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《中国临床神经外科杂志》[ISSN:1009-153X/CN:42-1603/TN]

卷:
期数:
2019年10期
页码:
608-610
栏目:
论著
出版日期:
2019-10-20

文章信息/Info

Title:
Effect of tea polyphenol on intracellular calcium concentration of primary hippocampal neurons and its mechanism in mice
文章编号:
1009-153X（2019）10-0608-03
作者:
严飞平　陈　晨　李志磊　霍国进　冯　毅　白　磊
719000 陕西，榆林市第一医院榆林院区神经外科（严飞平、陈　晨、李志磊、霍国进、冯　毅、白　磊）
Author(s):
YAN Fei-ping;  CHEN Chen;  LI Zhi-lei;  HUO Guo-jin;  FENG Yi;  BAI Lei.
Department of Neurosurgery, Yulin Municipal First Hospital, Yulin 719000, China
关键词:
小鼠海马神经元; 原代培养; 茶多酚; 细胞内钙离子浓度
Keywords:
Tea polyphenols;  Ischemia reperfusion injury;  Mice;  Primary hippocampal neurons;  intracellular calcium concentration
分类号:
R 743.3; Q 786
DOI:
10.13798/j.issn.1009-153X.2019.10.011
文献标志码:
A
摘要:
目的 探讨茶多酚对氧葡萄糖剥夺/再恢复干预后原代培养的小鼠海马神经元内钙离子浓度的影响及机制。方法 分离C57BL/6J雄性小鼠海马神经元并进行原代培养，然后分为对照组、氧葡萄糖剥夺/再恢复干预（模型）组和茶多酚组。对照组常规进行海马神经元原代培养，模型组利用氧葡萄糖剥夺/再恢复制作模型，茶多酚组在造模后应用4 mg/ml的茶多酚处理。利用Fluo-3AM钙离子特异性探针标记神经元内游离钙离子，利用流式细胞仪进行检测钙离子浓度；免疫印迹法检测神经元Cav1.2蛋白的表达水平；利用全细胞模式膜片钳技术检测神经元L-型电压依赖型钙离子通道（LTCC）的电流。结果 与对照组相比，氧葡萄糖剥夺/再恢复干预后，神经元内钙离子浓度明显增高（P<0.05），神经元Cav1.2蛋白表达水平明显增高（P<0.05）、LTCC电流明显增大（P<0.05）；茶多酚明显抑制上述作用（P<0.05），但未恢复到对照组水平（P<0.05）。结论 茶多酚可减少氧葡萄糖剥夺/再恢复干预后原代培养小鼠海马神经元内钙离子，其作用机制可能与降低Cav1.2表达、减弱LTCC功能有关。
Abstract:
Objective To investigate the effect of tea polyphenol on the intracellular calcium concentration of primary hippocampal neurons and its mechanism in the mice. Methods The primary hippocampal neurons were isolated from the brain of C57BL/6J male mice and randomly divided into control group, in which the hippocampal neurons were cultured in the conventional incubator, ischemia reperfusion group, in which the hippocampal neurons were cultured by oxygen-glucose deprivation/reperfusion method simulating ischemia reperfusion injury, and tea polyphenols group, in which the hippocampal neurons were treated with 4 mg/ml tea polyphenol 3 hours after incubation in the hypoxic incubator, and then cultured in the conventional incubator. The intracellular calcium concentration, expression of Cav1.2 and function of L-type calcium channel in the hippocampal neurons were determined in all the groups respectively by flow cytometry, Western blot and Whole-cell model. Results The intracellular calcium concentration, level of Cav1.2 expression and function of L- type calcium channel in the hippocampal neurons were significantly lower in the control group than those in tea polyphenol group (P<0.05), which were significantly lower than those in the ischemia reperfusion group (P<0.05). Conclusion It is suggested that tea polyphenols may decreases the intracellular calcium concentration of the primary hippocampal neurons probably by inhibiting the level of Cav1.2 expression and the function of L-type calcium channel in the mice with ischemia repekusion injury.

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备注/Memo

备注/Memo:
通讯作者：白　磊，E-mail：bailei2211@163.com（2019-06-13收稿，2019-09-05修回）
更新日期/Last Update: 2019-10-20