内源性大麻素对热应激反应的调节效应

发布时间：2018-05-26 15:13

  本文选题：内源性大麻素 + 大麻素受体　； 参考：《第三军医大学》2009年硕士论文

【摘要】：目的:内源性大麻素通过激活G蛋白耦连的大麻素受体(CB1和CB2)而发挥对神经、心血管、免疫等系统的调节作用。此外,内源性大麻素也具有抗炎、镇痛等多种医疗用途,但其生理效应尚未完全揭示。热应激反应是细胞最重要的应激保护机制之一,主要通过激活热休克转录因子1而显著增强包括分子伴侣机制在内的多种保护机制。而内源性大麻素对热应激反应有无调节效应及其机制如何,迄今国内外尚无研究报道。本研究在文献调研的基础上,初步推测大麻素可能通过其受体对热应激反应产生调节作用。迄今国内外尚无研究报道。本研究在文献调研的基础上,初步推测大麻素可能通过其受体对热应激反应产生调节作用。为证实这一假设,本研究拟以大鼠胶质瘤细胞株作为研究对象,观察内源性大麻素是否对其热应激反应具有调节效应,并探索其相应的受体机制;以期初步阐明内源性大麻对热应激反应的调节效应及机制,为内源性大麻素的效应及机制研究提供新思路和突破点。 方法:选用大鼠胶质瘤C6细胞为实验模型;实验组共分4类,即热应激组(42℃水浴1h)、热应激+内源性大麻素(2-AG)处理组、热应激+2-AG + CB1受体特异性拮抗剂(AM251)或CB2受体特异性拮抗剂(AM630)处理组、以及各自的对照组;设3h、6h、12h、24h等4个时相点。①采用RT-PCR技术,检测热应激、热应激+2-AG、热应激+2-AG+大麻素受体抑制剂处理后HSP70 mRNA表达水平。②采用Western Blot技术,检测热应激、热应激+2-AG、热应激+2-AG+抑制剂处理后HSP70蛋白表达水平。 结果:①C6细胞热应激处理后,分别于3h、6h、12h、24h检测HSP70 mRNA及蛋白的变化。结果显示,热刺激后3h、6h、12h、24h,HSP70 mRNA和蛋白表达水平均较对照组显著增高,12h时增加最为显著。②选择热应激后12h为观察时相点,以10-7mol/L、10-6mol/L、10-5mol/L三种不同浓度的2-AG进行处理,结果发现10-5mol/L 2-AG对热刺激1h后HSP70 mRNA、蛋白表达具有显著促进作用;而10-7mol/L、10-6mol/L2-AG则无此促进效应。③观察热刺激及10-5mol/L 2-AG处理后,不同时相点HSP70 mRNA和蛋白的表达情况,结果发现2-AG处理后HSP70 mRNA和蛋白表达在6h到达高峰,无2-AG处理对照组HSP70 mRNA和蛋白在12h到达高峰。④以CB1受体特异性拮抗剂(AM251)和CB2受体特异性拮抗剂(AM630)处理细胞后,检测2-AG对HSP70 mRNA、蛋白表达的影响,结果显示,AM251阻断了2-AG对HSP70 mRNA和蛋白的促进效应,而AM630无此效应。 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可以参与调控细胞热应激反应;此促进效应可被CB1特异性受体抑制剂AM251所抑制,表明2-AG的抑制效应是由CB1,而非CB2,信号通路所介导。本研究结果揭示了2-AG可能具备调控热应激效应的能力。
[Abstract]:Aim: endogenous cannabinoids regulate the nervous, cardiovascular and immune systems by activating the G protein-coupled cannabinoid receptors CB1 and CB2. In addition, endogenous cannabinoids also have a variety of anti-inflammatory, analgesic and other medical uses, but its physiological effects have not been fully revealed. Heat stress response is one of the most important mechanisms of cell stress protection, mainly through the activation of heat shock transcription factor 1 to significantly enhance a variety of protective mechanisms, including molecular chaperone mechanism. So far, there are no studies on whether endogenous cannabinoids regulate the thermal stress response and how they are related to them. On the basis of literature review, this study preliminarily speculated that cannabinoid may regulate the thermal stress reaction through its receptor. So far, there is no research report at home and abroad. On the basis of literature review, this study preliminarily speculated that cannabinoid may regulate the thermal stress reaction through its receptor. In order to confirm this hypothesis, this study intends to investigate whether endogenous cannabinoids can regulate the heat stress response of rat glioma cells and explore its receptor mechanism. The aim of this study was to elucidate the regulatory effect and mechanism of endogenous cannabis on heat stress and provide new ideas and breakthrough points for the study of endogenous cannabinoid effect and mechanism. Methods: rat glioma C6 cells were selected as the experimental model, the experimental group was divided into 4 groups: heat stress group (42 鈩，

本文编号：1937718