摘要
目的探讨17-AAG在氧糖剥夺后复糖复氧(OGD/R)诱导的神经元凋亡中的保护作用及可能的机制。方法体外培养大鼠原代神经元并建立OGD/R模型。通过TUNEL实验分别检测氧糖剥夺0.5、1、2h复糖复氧24h后的神经元凋亡情况;通过TUNEL实验检测不同浓度17-AAG(0.5、1、2μmol/L)对OGD/R处理后神经元凋亡的影响;采用Western blotting检测17-AAG对OGD/R处理后神经元中HSP70蛋白表达的影响;构建HSP70干扰慢病毒,通过TUNEL实验检测HSP70干扰在17-AAG调节OGD/R诱导的神经元凋亡中的作用。结果 OGD/R能够明显诱导神经元的凋亡,且神经元凋亡率随着氧糖剥夺时间的增加而升高。17-AAG能够明显抑制OGD/R诱导的神经元凋亡,浓度越高抑制作用越明显;OGD/R能够明显抑制神经元中HSP70蛋白的表达,而17-AAG则能够明显逆转OGD/R对神经元中HSP70表达的抑制作用;HSP70慢病毒干扰可明显逆转17-AAG对OGD/R处理神经元的保护作用。结论 17-AAG通过上调HSP70的表达来抑制OGD/R诱导的神经元凋亡。
Objective To explore the role and mechanism of 17-allylamino-17-demethoxygeldanamycin（17-AAG） in neurons apoptosis induced by oxygen-glucose deprivation and recovery（OGD/R）. Methods Primary rat neurons were cultivated in vitro, and OGD/R model was reproduced. Neurons were exposed to OGD for 0.5 h, 1 h and 2 h, and then reperfusion for 24 h, the effect of OGD/R on neurons apoptosis was detected by TUNEL assay. On OGD/R, neurons were treated with different concentrations of 17-AAG（0.5, 1.0 and 2.0μmol/L）, and the effect of 17-AAG on OGD/R treated neurons apoptosis was detected by TUNEL assay. Western blotting was performed to detect the expression of heat shock protein 70（HSP70）. HSP70 interference lentivirus was then constructed. The effect of HSP70 interference on the neurons apoptosis treated with OGD/R and 17-AAG was detected by TUNEL assay. Results OGD/R significantly induced neurons apoptosis, and the rate of neurons apoptosis increased with the increase of OGD time. 17-AAG obviously inhibited the neurons apoptosis induced by OGD/R, and the higher the 17-AAG concentration, the more obvious the inhibition. OGD/R significantly suppressed the expression of HSP70 protein in neurons, and 17-AAG obviously reversed the inhibition of HSP70 protein expression induced by OGD/R. However, HSP70 lentivirus interference markedly reversed the protective effect of 17-AAG on OGD/R treated neurons. Conclusion 17-AAG may inhibit the apoptosis of OGD/R treated neurons by up-regulating HSP70.
作者
李建雄
李鸣明
卜玉洁
李斌
邹华
张廷华
LI Jian-xiong;LI Ming-ming;BU Yu-jie;LI Bin;ZOU Hua;ZHANG Ting-hua(Department of Neurologyj Second Hospital of Lanzhou University, Lanzhou 730030, Chin)
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2018年第4期310-315,共6页
Medical Journal of Chinese People's Liberation Army
基金
甘肃省兰州市科技计划项目基金(2017-2-70)~~
