大鼠蛛网膜下腔出血后脑组织socs3的表达研究

发布时间：2018-05-03 03:02

本文选题：蛛网膜下腔出血 + 细胞因子信号转导负调控因子3　；参考：《南京大学》2014年硕士论文

【摘要】：研究背景及目的蛛网膜下腔出血(SAH)是一种严重的神经外科疾病。蛛网膜下腔出血后早期脑损伤(EBI)在迟发性脑缺血的发生发展中起到重要作用,并且是造成患者高致残率及致死率的首要原因。蛛网膜下腔出血后早期脑组织发生氧化应激、免疫炎症反应、细胞凋亡等病理生理改变。JAK/STAT信号通路参与多种信号通路级联反应,细胞因子信号转导负调控因子3(SOCS3)通过JAK/STAT通路调节信号传导。由此考虑在蛛网膜下腔出血后的早期脑损伤,SOCS3参与其生理病理过程,并发挥调节信号转导作用。本实验研究在蛛网膜下腔出血(SAH)后早期脑组织中SOCS3的分布和表达变化,及与炎性细胞因子相关性。方法成年雄性SD大鼠72只(250-300g,南京军区总医院动物中心提供),分为对照组、SAH后3h、 6h、12h、24h、 48h、72h、5d、7d共9组(每组8只)。采用视交叉池注血法建立大鼠SAH模型,于SAH后不同时间点处死后取脑,应用免疫印迹和免疫组化技术检测SAH后脑组织中SCS3的分布和表达变化,并通过实时荧光定量聚合酶链反应(Realtime-PCR)检测炎性因子IL-1β及TNF-α表达及变化。结果SAH后大鼠脑组织中3h时SOCS3蛋白水平开始升高,24h达到高峰,其后逐渐下降,7d仍高于对照组。SOCS3广泛存在于神经系统细胞中,其中神经元与部分胶质细胞均有较明显SOCS3阳性表达。同时,对照组中SOCS 3主要定位于胞核,SAH后SOCS3胞浆阳性增加,24h达到高峰,其后胞核胞浆阳性都下降。Realtime-PCR示SAH后炎性介质IL-1β表达显著增加,3h后开始升高,1d达第一个高峰,5d达到表达的第二个高峰,CNF-α于3h达到峰值,后逐渐减少。结论大鼠SAH后早期伴有SOCS3信号通路的激活,提示SOCS3信号通路参与SAH后早期脑损伤中的病理生理改变。炎性因子IL-1β及TNF-a参与大鼠SAH后脑损伤生理病理过程,可能通过SOCS3发挥作用。
[Abstract]:Background and objective Subarachnoid hemorrhage (SAH) is a serious neurosurgical disease. Early brain injury (EBI) after subarachnoid hemorrhage plays an important role in the occurrence and development of delayed cerebral ischemia and is the primary cause of high disability rate and fatality rate. Oxidative stress, immune inflammation, apoptosis and other pathophysiological changes in early brain tissue after subarachnoid hemorrhage. Cytokine signal transduction negative regulator 3 (SOCS 3) regulates signal transduction through JAK/STAT pathway. It is considered that early brain injury after subarachnoid hemorrhage may participate in the physiological and pathological process of SOCS3 and play a role in regulating signal transduction. The present study was designed to investigate the distribution and expression of SOCS3 in brain tissue after subarachnoid hemorrhage (SAH) and its correlation with inflammatory cytokines. Methods Seventy-two adult male Sprague-Dawley rats were divided into 9 groups (n = 8 in each group). They were divided into 9 groups (n = 8 in each group), which were provided by the Animal Center of Nanjing military region General Hospital for 3 h, 6 h, 12 h, 24 h, 48 h, 72 h, 5 d and 7 d after SAH respectively. The rat SAH model was established by injecting blood into the cistern of optic chiasma. The brain was killed at different time points after SAH. The distribution and expression of SCS3 in brain tissue after SAH were detected by immunoblotting and immunohistochemistry. The expression and change of inflammatory factor IL-1 尾 and TNF- 伪 were detected by real-time fluorescence quantitative polymerase chain reaction (PCR). Results the level of SOCS3 protein began to increase at 3 h after SAH and reached its peak at 24 h, and then decreased gradually at 7 days, which was still higher than that in the control group. SOCS3 was widely expressed in nervous system cells, and the positive expression of SOCS3 in neurons and some glial cells was obvious. At the same time, in the control group, SOCS 3 was mainly located in the nucleus of the cell, and the increase of SOCS3 cytoplasmic positive rate reached its peak at 24 h. After that, the expression of IL-1 尾 in the cytoplasm decreased. Realtime-PCR showed that the expression of IL-1 尾 in the inflammatory mediators increased significantly after SAH for 3 h, and the expression of IL-1 尾 began to increase at 1 day and reached the first peak at 5 days. The second peak of CNF- 伪 reached its peak at 3 h, and then decreased gradually. Conclusion the activation of SOCS3 signal pathway after SAH in rats suggests that SOCS3 signaling pathway is involved in the pathophysiological changes of early brain injury after SAH. Inflammatory factors IL-1 尾 and TNF-a participate in the physiological and pathological process of brain injury after SAH in rats, which may play a role through SOCS3.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.35

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