缺血再灌注损伤大鼠脑组织及外周血炎症细胞因子分析
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摘要
目的探讨大鼠脑缺血再灌注损伤后6 h和24 h时炎症细胞因子在脑组织和血清中的变化,初步分析炎症因子在早期缺血再灌注损伤中的作用。方法构建大鼠脑中动脉闭塞模型,运用流式高通量多因子检测方法测定各因子变化。结果缺血再灌注6 h,脑组织中促炎细胞因子IL-6、IFN-γ、MIP-2、TNF-α、IL-1β均升高;抗炎细胞因子IL-10升高,差异均有统计学意义(P<0.05)。缺血再灌注24 h,促炎细胞因子IL-6、MIP-2、IL-1β升高。缺血再灌注24 h较6 h促炎细胞因子IL-6、IL-1β升高,IFN-γ、MIP-2降低,差异均有统计学意义(P<0.05)。血清中,缺血再灌注6 h和24 h促炎细胞因子IL-6、IFN-γ、MIP-2、TNF-α、IL-1β均升高。缺血再灌注24 h,抗炎细胞因子IL-10、IL-13升高。缺血再灌注24 h较6 h促炎细胞因子IL-6、MIP-2、TNF-α、IL-1β升高,IFN-γ降低;抗炎细胞因子IL-13持续升高,差异均有统计学意义(P<0.05)。结论血中炎症因子的变化可作为脑缺血再灌注的血清标志物。
Objective To investigate the changes of inflammatory cytokines in brain tissue and serum at 6 h and 24 h respectively after cerebral ischemia-reperfusion(I/R) injury in rats. To conduct a preliminary analysis of how inflammatory cytokines act in early ischemia-reperfusion injury. Methods To set up the rat middle cerebral artery occlusion(MCAO) model, and the changes of each factor were measured by multiple immunoassays for flow test. Results In I/R 6 h, the levels of pro-inflammatory cytokines including IL-6, IFN-γ, MIP-2, TNF-α and IL-1β increased in brain tissue, so does anti-inflammatory cytokines IL-10, with the differences statistically significant(P<0.05). In I/R 24 h, the pro-inflammatory cytokines including IL-6, MIP-2, IL-1β increased. Compared with I/R at 6 h, pro-inflammatory cytokines IL-6, IL-1β increased whereas IFN-γ, MIP-2 decreased in I/R 24 h, with the differences statistically significant(P<0.05). In serum, the levels of pro-inflammatory cytokines including IL-6, IFN-γ, MIP-2, TNF-α and IL-1β increased in both I/R 6 h and 24 h. In the I/R 24 h, anti-inflammatory cytokines IL-10 and IL-13 increased. Compared with I/R 6 h, pro-inflammatory cytokines IL-6, MIP-2, TNF-α, IL-1β increased and IFN-γ decreased whereas anti-inflammatory IL-13 increased continuously in I/R 24 h, with the differences statistically significant(P<0.05). Conclu￣sion The changes of inflammatory cytokines in peripheral blood can be used as serum markers of cerebral ischemia-reperfusion.
Objective To investigate the changes of inflammatory cytokines in brain tissue and serum at 6 h and 24 h respectively after cerebral ischemia-reperfusion(I/R) injury in rats. To conduct a preliminary analysis of how inflammatory cytokines act in early ischemia-reperfusion injury. Methods To set up the rat middle cerebral artery occlusion(MCAO) model, and the changes of each factor were measured by multiple immunoassays for flow test. Results In I/R 6 h, the levels of pro-inflammatory cytokines including IL-6, IFN-γ, MIP-2, TNF-α and IL-1β increased in brain tissue, so does anti-inflammatory cytokines IL-10, with the differences statistically significant(P<0.05). In I/R 24 h, the pro-inflammatory cytokines including IL-6, MIP-2, IL-1β increased. Compared with I/R at 6 h, pro-inflammatory cytokines IL-6, IL-1β increased whereas IFN-γ, MIP-2 decreased in I/R 24 h, with the differences statistically significant(P<0.05). In serum, the levels of pro-inflammatory cytokines including IL-6, IFN-γ, MIP-2, TNF-α and IL-1β increased in both I/R 6 h and 24 h. In the I/R 24 h, anti-inflammatory cytokines IL-10 and IL-13 increased. Compared with I/R 6 h, pro-inflammatory cytokines IL-6, MIP-2, TNF-α, IL-1β increased and IFN-γ decreased whereas anti-inflammatory IL-13 increased continuously in I/R 24 h, with the differences statistically significant(P<0.05). Conclu￣sion The changes of inflammatory cytokines in peripheral blood can be used as serum markers of cerebral ischemia-reperfusion.
引文
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[15] Daines MO, Hershey GK. A novel mechanism by which interferon-gamma can regulate interleukin (IL)-13 responses. Evidence for intracellular stores of IL-13 receptor alpha -2 and their rapid mobilization by interferon-gamma[J]. J Biol Chem, 2002, 277(12): 10387-10393.
[2] Li WX, Qi F, Liu JQ, et al. Different impairment of immune and inflammation functions in short and long-term after ischemic stroke[J]. Am J Transl Res, 2017, 9(2): 736-745.
[3] Singh N, Agrawal M, Dore S. Neuroprotective properties and mechanisms of resveratrol in in vitro and in vivo experimental cerebral stroke models[J]. Acs Chem Neurosci, 2013, 4(8): 1151-1162.
[4] Longa EZ, Weinstein PR, Carlson S, et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke, 1989, 20(1): 84-91.
[5] Lopez MS, Dempsey RJ, Vemuganti R. Resveratrol neuroprotection in stroke and traumatic CNS injury[J]. Neurochem Int, 2015, 89: 75-82.
[6] Nakka VP, Prakash-babu P, Vemuganti R. Crosstalk between endoplasmic reticulum stress, oxidative stress, and autophagy: Potential therapeutic targets for acute CNS injuries[J]. Mol Neurobiol, 2016, 53(1): 532-544.
[7] Erta M, Quintana A, Hidalgo J. Interleukin-6, a major cytokine in the central nervous system[J]. Int J Biol Sci, 2012, 8(9): 1254-1266.
[8] Brea D, Sobrino T, Ramos-Cabrer P, et al. Inflammatory and neuroimmunomodulatory changes in acute cerebral ischemia[J]. Cerebrov Dis, 2009, 27(Suppl 1): 48-64.
[9] Pei J, You X, Fu Q. Inflammation in the pathogenesis of ischemic stroke[J]. Front Biosci, 2015, 20: 772-783.
[10] Chuang DY, Simonyi A, Kotzbauer PT, et al. Cytosolic phospholipase A2 plays a crucial role in ROS/NO signaling during microglial activation through the lipoxygenase pathway[J]. J Neuroinflamm, 2015, 12: 199.
[11] Seifert HA, Collier LA, Chapman CB, et al. Pro-inflammatory interferon gamma signaling is directly associated with stroke induced neurodegeneration[J]. J Neuroimmune Pharm, 2014, 9(5): 679-689.
[12] Guo RB, Wang GF, Zhao AP, et al. Paeoniflorin protects against ischemia-induced brain damages in rats via inhibiting MAPKs/NF-kappaB-mediated inflammatory responses[J]. PLoS ONE, 2012, 7(11): e49701.
[13] Iadecola C, Anrather J. Stroke research at a crossroad: asking the brain for directions[J]. Nat Neurosci, 2011, 14(11): 1363-1368.
[14] Shin WH, Lee DY, Park KW, et al. Microglia expressing interleukin-13 undergo cell death and contribute to neuronal survival in vivo[J]. Glia, 2004, 46(2): 142-152.
[15] Daines MO, Hershey GK. A novel mechanism by which interferon-gamma can regulate interleukin (IL)-13 responses. Evidence for intracellular stores of IL-13 receptor alpha -2 and their rapid mobilization by interferon-gamma[J]. J Biol Chem, 2002, 277(12): 10387-10393.
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