褪黑素对大鼠脑缺血再灌注保护作用的研究

发布时间：2018-05-09 16:07

本文选题：褪黑素 + 脑缺血再灌注　；参考：《北京协和医学院》2017年博士论文

【摘要】：目的:探究褪黑素对大鼠MCAO术后脑缺血再灌注损伤的保护作用,并研究其具体保护机制。方法:36只大鼠随机分为假手术组(A组)、MCAO+溶剂组(B组)、MCAO+褪黑素组(C组)3组,分别给予颈部正中皮肤切开缝合、MCAO缺血2小时再灌注24小时+2次腹腔注射10%二甲基亚砜(1mL/kg)、MCAO缺血2小时再灌注24小时+2次腹腔注射褪黑素溶液(1mL/kg)。再灌注24小时后分别获取大脑标本、血清标本,对比各组大脑缺血再灌注损伤情况及氧化、炎症情况。结果:A组、B组和C组的神经功能行为学评分分别为0±0 vs.2.00±0.28 vs.1.33±0.14,C组相比于B组,神经功能行为学评分显著降低(p0.05)。TTC染色结果计算脑梗死体积百分比,A组、B组和C组分别为0±0 vs.66.05±8.89%vs.39.96±7.37%,C组相比于B组,脑梗死体积百分比显著降低(p0.05)。脑组织NF-κB免疫组化平均OD值,相比于A组,B组和C组显著升高(p0.01);C组相比于B组显著降低(p0.05)。脑组织IL-1β免疫组化平均OD值,相比于A组,B组和C组显著升高(P0.05);C组相比于B组显著降低(p0.05)。相比于A组,B组和C组的血清总SOD活性显著降低(p0.01);C组相比于B组显著升高(p0.01)。相比于A组的血清MDA含量,B组显著升高(P0.01),C组无统计学差异(p=0.1052);C相比于B组显著降低(p0.05)。脑组织氧化、炎症相关损伤因子的蛋白水平检测结果显示,相比于B组,C组IL-1β、NF-κB、iNOS、S100β表达量显著降低,Nrf-2表达量显著升高(p0.05);基因水平检测结果显示,C组较B组IL-1β、NF-κB、iNOS表达量显著降低,Nrf-2表达量显著升高(p0.05)。进一步行炎性小体相关通路研究,蛋白水平检测结果显示C组NLRC4、ASC、CASP-1表达量较B组显著降低(p0.05),AIM2表达量无明显差异(p=0.4901);基因水平检测结果显示,C组相比于B组,NLRC4、ASC、CASP-1基因表达量显著降低(p0.05),而两组AIM2表达量无明显差异(p=0.9884)。结论:大鼠MCAO术后会导致的脑缺血再灌注损伤,褪黑素通过抗氧化和抗炎等方式减轻脑缺血再灌注损伤。褪黑素可能通过与NLRC4炎性小体相互作用,调节脑缺血再灌注相关炎症反应。
[Abstract]:Objective: To explore the protective effect of melatonin on cerebral ischemia reperfusion injury after MCAO operation in rats, and to study the specific protective mechanism. Methods: 36 rats were randomly divided into sham operation group (group A), MCAO+ solvent group (group B), MCAO+ melatonin group (group C), and group of 3 groups were treated with normal neck skin incision and suture, MCAO ischemia 2 hours reperfusion for 24 hours +2 abdominal cavity Injection of 10% two methyl sulfoxide (1mL/kg), MCAO ischemia for 2 hours and reperfusion for 24 hours and +2 times, and intraperitoneal injection of melatonin solution (1mL/kg). After reperfusion for 24 hours, the brain specimens and serum specimens were obtained respectively. The cerebral ischemia reperfusion injury and oxidation and inflammation were compared in each group. The neurological functional behavior score of A group, B group and C group was 0 +. 0 vs.2.00 + 0.28 vs.1.33 + 0.14, C group compared with B group, the neurological functional behavior score was significantly reduced (P0.05).TTC staining results to calculate the percentage of cerebral infarction volume, A group, B group and C group were 0 0 vs.66.05 + 8.89%vs.39.96 7.37% respectively, C group compared with the group, cerebral infarction volume ratio significantly decreased. Compared with group A, group B and C group were significantly higher (P0.01), and C group was significantly lower than group B (P0.05). The average OD value of IL-1 beta immunohistochemistry in brain tissue was significantly higher than that of A group, B group and C group (P0.05). Higher (P0.01). Compared with the serum MDA content in the A group, the B group increased significantly (P0.01), and there was no significant difference in the C group (p=0.1052); C was significantly lower than the B group (P0.05). The protein level of the brain tissue was oxidized and the protein level of the inflammatory related damage factors showed that the expression of beta, kappa, and beta expression was significantly lower than that in the B group. (P0.05), the results of gene level detection showed that the expression of IL-1 beta, NF- kappa B, iNOS and Nrf-2 expression were significantly lower in group C than in group B, and the expression of Nrf-2 was significantly increased (P0.05). The results of level detection showed that the expression of NLRC4, ASC and CASP-1 was significantly lower in group C than in B group (P0.05), but there was no significant difference in the expression of AIM2 in the two groups (p=0.9884). Conclusion: melatonin can reduce cerebral ischemia and reperfusion injury in rats after MCAO operation. Melatonin may pass through the effects of antioxidation and anti inflammation. NLRC4 inflammatory corpuscle interaction regulates cerebral ischemia-reperfusion related inflammatory response.

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R743

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