内源性大麻素2-AG促进HSP70基因的热应激表达

Endocannabinoid 2-AG enhances HSP70 gene expression in heat stress response in vitro

目的探讨内源性大麻素对热应激反应是否存在调控作用,及其可能的受体调控机制。方法以不同浓度(10-7、10-6、10-5mol/L)内源性大麻素2-AG(2-Arachidonylglycerol)预处理大鼠胶质瘤C6细胞1h,热刺激(42℃水浴)1h,通过RT-PCR及Western blot检测热休克蛋白70(heat shock protein70,HSP70)的mRNA及蛋白表达水平;同时观察特异性受体拮抗剂AM251或AM630的拮抗效应,探讨信号通路机制。结果内源性大麻素2-AG可显著促进热刺激处理的C6细胞HSP70基因及蛋白的表达,该调控效应由CB1介导。结论一定浓度的内源性大麻素2-AG对热刺激处理的C6细胞内HSP70基因的表达有明显促进效应,表明2-AG对细胞热应激反应可能具有一定的调控能力;同时,在C6细胞内,该调控效应主要由CB1介导。

Objectives To investigate the regulation role of endocannabinoid in heat stress response and its possible receptor signaling pathway. Methods Rat glioma C6 cells were pretreated with different concentrations （10^-7, 10^-6 and 10^-5mol/L） of endocannabinoid 2-Arachidonylglycerol （2-AG） for 1 h, then heat stress （42℃ water bath） was performed for t h. Reverse transeription-PCR and Western blotting were used to detect the expression levels of HSP70 mRNA and protein. The blockade effects of selective CB1 antagonist AM251 （10^-4mol/L） or CB2 antagonist AM630 （10^-4mol/L） were also observed by adding them to the culture medium when 2-AG was pretreated. Resolts The expression levels of HSP70 mRNA and protein in heat stressed C6 eells were significantly increased after 2-AG treatment. This enhancement was mainly through CB1 receptor signaling pathway. Gonolusion Certain level of 2-AG significantly promotes the expression of HSP70 gene post heat stress, suggesting that 2-AG is involved in the regulation of heat stress response. The promotion effect of 2-AG in C6 cells is mainly mediated through CB1 receptor signaling pathway, but not CB2.