胆碱能抗炎通路对大鼠脑缺血再灌注损伤的影响

发布时间：2018-04-14 09:16

本文选题：胆碱能抗炎通路 + 脑缺血再灌注　；参考：《北京市结核病胸部肿瘤研究所》2008年硕士论文

【摘要】： 目的:应用侧脑室注射M1型胆碱能受体激动剂McN-A-343、M2型胆碱能受体拮抗剂美索曲明(Methoctramine)以及α7亚单位的N型胆碱能受体激动剂胆碱(Choline)的方法激活胆碱能抗炎通路(cholinergic anti-inflammatory pathway,CAP),通过检测大鼠左侧海马、心、肝、肺、肾和血浆中肿瘤坏死因子-α(tumor necrosis factor alpha, TNF-α)、白介素-1β(interleukin-1β,IL-1β)含量及右侧海马CA1区细胞凋亡数,进一步探讨CAP对大鼠脑缺血再灌注损伤的影响。方法:健康成年雄性Sprague-Daewley大鼠25只,随机分为5组(n=5):假手术组(SHAM组)、脑缺血再灌注组(I/R组)、美索曲明组(MET组)、McN-A-343组(MA343组)和胆碱组(CHO组)。4-VO[1]法建立全脑缺血再灌注损伤模型,电凝大鼠双侧椎动脉(SHAM组除外),于缺血前15min分别经侧脑室向MET组、MA343组及CHO组注射Methoctramine(500ng/kg,10μl)、McN-A-343(500ng/kg,10μl)及Choline(500ng/kg,10μl),SHAM组及I/R组注射生理盐水10μl。除SHAM组外,各组夹闭双侧颈总动脉20min后再灌注。再灌注6h后,断头处死动物,采集标本。放射免疫法(RIA)检测左侧海马、心、肝、肺、肾和血浆中TNF-α、IL-1β含量。TdT介导的dUTP缺口末端标记技术(Terminal deoxynucleotidyl transferase fluorescein-12-dUTP nick end labeling, TUNEL)染色检测右侧海马CA1区细胞凋亡数。结果:再灌注后,MET组和MA343组海马、心、肝、肾组织匀浆及血浆TNF-α、IL-1β含量显著低于I/R组(P0.05),肺组织TNF-α、IL-1β含量差异无统计学意义(P0.05)。CHO组海马TNF-α、IL-1β含量较I/R组有所降低(P0.05),而心、肺、肝、肾及血浆TNF-α、IL-1β含量差异无统计学意义。与I/R组相比,MET、MA343及CHO组海马CA1区凋亡细胞数目减少(P0.05)。结论:TNF-α、IL-1β的释放是脑缺血再灌注损伤的重要机制之一,CAP对脑缺血再灌注引发的局部及全身炎症反应均具有保护作用,对细胞凋亡有间接抑制作用。其机制与Methoctramine、McN-A-343和Choline激活CAP后抑制缺血再灌注TNF-α、IL-1β的释放有关。
[Abstract]:Objective: to activate the cholinergic anti-inflammatory pathwayCAPPAP pathway by intracerebroventricular injection of the M1 type cholinergic receptor agonist McN-A-343M 2 cholinergic receptor antagonist methoctramine and the N-type cholinergic receptor agonist choline of 伪 7 subunit.By detecting the left hippocampus of rats,The contents of TNF- 伪 tumor necrosis factor alpha, TNF- 伪, interleukin-1 尾 interleukin-1 尾 interleukin-1 尾 IL-1 尾 in heart, liver, lung, kidney and plasma and the number of apoptosis in the right hippocampal CA1 region were studied to investigate the effect of CAP on cerebral ischemia-reperfusion injury in rats.Methods: Twenty-five healthy adult male Sprague-Daewley rats were randomly divided into 5 groups: sham-operated group (sham group), cerebral ischemia-reperfusion group (I / R) group, misotromine group (MET group) and choline group (Cho group) and choline group (Cho group. 4-VO [1] method) to establish global cerebral ischemia-reperfusion injury model.Before ischemia, 15min was injected into MET group (MA343 group) and CHO group (10 渭 g / kg), Choline500ngSHAM group (10 渭 l) and Choline 500ng / kg SHAM group (10 渭 l) and I / R group (10 渭 l).With the exception of SHAM group, 20min of bilateral common carotid artery was clipped and reperfusion in each group.After reperfusion for 6 hours, the animals were killed and the specimens were collected.The terminal deoxynucleotidyl transferase fluorescein-12-dUTP nick end labeling (Tunel) in left hippocampus, heart, liver, lung, kidney and plasma mediated by terminal deoxynucleotidyl transferase fluorescein-12-dUTP nick end labeling (Tunel) were used to detect the apoptosis of CA1 in the right hippocampus.Results: after reperfusion, the contents of TNF- 伪 IL-1 尾 in hippocampus, heart, liver, kidney homogenate and plasma in the met group and the MA343 group were significantly lower than those in the I / R group (P 0.05). There was no significant difference in the content of TNF- 伪 and IL-1 尾 between the two groups (P 0.05), but the contents of TNF- 伪 IL-1 尾 in the heart, lung, liver, liver, and lung were significantly lower than those in the I / R group.There was no significant difference in the contents of TNF- 伪 and IL-1 尾 between kidney and plasma.Compared with the I / R group, the number of apoptotic cells in the hippocampal CA1 region of CHO group was significantly lower than that of METMA343 group.Conclusion the release of IL-1 尾 from TNF- 伪 is one of the important mechanisms of cerebral ischemia-reperfusion injury. CAP has protective effect on local and systemic inflammation induced by cerebral ischemia-reperfusion, and indirectly inhibits apoptosis.The mechanism is related to the inhibition of TNF- 伪 IL-1 尾 release after activation of CAP by method minefield McN-A-343 and Choline.
【学位授予单位】：北京市结核病胸部肿瘤研究所
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R741

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