枸杞多糖对氧糖剥夺再灌注损伤后海马神经元凋亡与自噬性死亡的作用  

Effect of lycium barbarum polysaccharides of apoptosis and autophagic death in primary cultured hippocampul neurons injured by oxygen glucose deprivation and reoxygenation

作　　者：鱼洋 李星慧 濮璟楠 武秀权 罗鹏[1] 王玖[1] 费舟[1] Yang;Li Xinghui;Pu Jingnan;Wu Xiuquan;Luo Peng;Wang Jiu;Fei Zhou(Department of Neurosurgery,Xijing Hospital,Fourth Military Medical University,Xi＇an 710032,China;School of Public Health,Shanxi University of Chinese Medicine,Xianyang 712046,China)

机构地区：[1]第四军医大学西京医院神经外科,西安710032 [2]陕西中医药大学公共卫生学院,咸阳712046

出　　处：《中华神经医学杂志》2018年第7期649-655,共7页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（8143000089、81771239、81771322）

摘　　要：目的探究枸杞多糖（LBP）对原代海马神经元氧糖剥夺再灌注（OGD／R）损伤的保护作用及对细胞凋亡与自噬性细胞死亡的调节作用。方法将原代培养的小鼠海马神经元分为5组，对照组（不做OGD／R处理）、0GD瓜组和OGD／R＋LBP15wg／mL组、0GD／R＋LBP30μg／mL组、OGD／R＋LBP60wg／mL组。采用MTT法检测细胞存活率，LDH释放率测定细胞损伤程度，TUNEL染色法检测凋亡率，免疫荧光染色法观察活化型Caspase．3定位，Westernblotting法测定蛋白表达量。结果OGD／R损伤后，细胞存活率显著降低，而不同浓度LBP（15、30、60wg／mL）处理可明显减轻细胞损伤并降低LDH漏出率。TUNEL染色法检测显示OGD／R＋LBP60μg／mL组细胞阳性凋亡数明显降低。免疫荧光染色法和Western blotting结果显示，与OGD／R组相比。OGD肘LBP60wg／mL组活化型Caspase-3／Caspase-3比值、LC3Ⅱ／LC3Ⅰ比值、Beclin1蛋白表达明显减低，Bcl-2／Bax比值、p62蛋白表达明显增加，差异均有统计学意义（P〈0．05）。结论LBP通过抑制细胞凋亡与自噬性细胞死亡对OGD瓜诱导的海马神经元损伤起保护作用。Objective To investigate the effect of lycium barbarum polysaccharides （LBP） on apoptosis and autophagic death in primary cultured hippocampal neurons after oxygen glucose deprivation and reoxygenation （OGD/R） injury. Methods Primary cultured hippocampal neurons were exposed to OGD/R. The ceils were randomly divided into 5 groups： control group, OGD/R group, and OGD/R＋LBP groups （15, 30 and 60 μg/mL LBP）. The cell viability was assessed by MTT assay. The cell damage was evaluated through detecting the lactate dehydrogenase （LDH） release rate. Apoptosis rate was detected by TUNEL, and cleaved Caspase-3 was identified by immunofluorescence staining. Protein expression was detected by Western blotting. Results As compared with the control group, OGD/R group had significantly decreased cell viability （P〈0.05）; and significantly increased cell viability and decreased LDH release rate were noted in the LBP （15, 30 and 60 μg/mL） treatment groups as compared with those in the OGD/R group （P〈0.05）. The 60 μg/mL LBP treatment group had significantly smaller number of TUNEL-positive cells than the OGD/R group（P〈0.05）. Immunofluorescence staining and Western blotting both revealed that 60 μg/mL LBP treatment group had significantly decreased Beclinl level and ratio of cleaved Caspase-3/Caspase-3 and ratio of mierotubule-associated protein light chain （LC）3IFLC3I, and statistically increased p62 level and ratio of Bcl-2/Bax as compared with OGD/R group （P〈0.05）. Conclusion LBP treatment protects primary hippocampal neurons from OGD/R injury via inhibiting apoptosis and autophagic cell death.

关 键 词：枸杞多糖 海马神经元 氧糖剥夺再灌注 自噬 细胞凋亡 

分 类 号：Q25[生物学—细胞生物学]