tBHQ对颅脑创伤后炎症反应保护作用的体外研究

发布时间：2018-01-03 18:16

本文关键词：tBHQ对颅脑创伤后炎症反应保护作用的体外研究　出处：《东南大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的：Nrf2通路在颅脑创伤后被激活,在颅脑创伤后具有极为重要的保护作用。动物实验证实,tBHQ作为Nrf2的激活剂,可以抑制小鼠颅脑创伤后的炎症反应,减轻继发性脑损伤。本研究拟通过体外实验,进一步探究tBHQ对颅脑创伤后炎症反应的保护作用及可能机制。方法：ICR胎鼠培养原代神经元细胞,待细胞成熟后,分为三组：(1)对照组；(2)TBI组：(3) TBI+tBHQ组。每组18孔细胞。处理结束24h后Western-Blot技术观察Nrf2胞浆/核含量,EMSA技术研究核内Nrf2的DNA结合活性、NF-κB的DNA结合活性；Real-timePCR技术定量测定Nrf2-ARE通路下游因子NQO1、HO-1、GST、γGCS的mRNA表达水平；化学发光法检测细胞上清GSH、GSSG含量；荧光发光法检测细胞上清ROS水平。结果：TBI后细胞核内Nrf2及下游因子NQO1、 HO-1、GST、γGCS表达增加,tBHQ可进一步增加其表达水平(P0.05)。TBI后NF-κB表达明显增加,细胞上清ROS水平上升,GSH含量下降(P0.05)。对比TBI组,tBHQ可显著抑制NF-κB的表达,降低细胞上清ROS水平(P0.05)。结论：tBHQ可抑制TBI后继发性炎症反应,从而起到保护作用。其机制可能是通过激活的Nrf2调控细胞内氧化还原反应,进而抑制NF-κB等炎症信号通路。
[Abstract]:Objective: Nrf2 pathway is activated in the brain after trauma, plays an important role in protecting the brain injury. Animal experiments confirmed that tBHQ as an activator of Nrf2, can inhibit the inflammatory response in mice after traumatic brain injury, reduce secondary brain injury. This study by in vitro experiments, to further explore the protective effect of tBHQ on the inflammatory response after traumatic brain injury and its possible mechanism. Methods: ICR rat primary cultured neurons, to mature cells, divided into three groups: (1) control group; (2) TBI group: (3) TBI+tBHQ group. Each of the 18 hole cell. Observed Nrf2 cytoplasmic / nuclear Western-Blot content after the end of 24h, EMSA technology to study the nuclear Nrf2 binding activity of DNA, NF- K B DNA binding activity; quantitative Real-timePCR determination of Nrf2-ARE downstream factor NQO1, HO-1, GST, GCS gamma mRNA expression; chemiluminescence detection of cell supernatant containing GSH, GSSG The amount of ROS in the cell supernatant; detection level of fluorescence method. Results: after TBI nucleus Nrf2 and downstream factor NQO1, HO-1, GST, gamma GCS expression increased, tBHQ can further increase the expression level of (P0.05) NF- K B expression was significantly increased after.TBI cell supernatant ROS levels increased, GSH content decreased (P0.05). Compared with TBI group, the expression of tBHQ could significantly inhibit NF- kappa B, reduce the level of ROS cells (P0.05). Conclusion: tBHQ can inhibit the secondary inflammatory reaction after TBI, which play a protective role. The mechanism is probably through the reaction of activated Nrf2 regulation of intracellular redox, and inhibition of NF- K B inflammation signal pathway.

【学位授予单位】：东南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.15

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