小鼠心肌缺血再灌注损伤后NF-κB磷酸化状态及炎症因子的表达水平

Phosphorylation of NF-κB and expression of inflammatory factors in myocardial ischemia-reperfusion injury in mice

目的探讨小鼠心肌缺血再灌注损伤后核因子-κB(nuclear factor kappa-B,NF-κB)磷酸化状态及炎症因子的表达水平.方法15只小鼠采用冠脉结扎再灌注法建立小鼠心肌缺血再灌注模型(观察组),另选15只小鼠作为对照组.再灌注损伤4h后,处死小鼠,HE染色分析心肌组织病理变化.取心肌组织采用Western blot分析NF-κB信号通路变化,采用酶联免疫吸附测定(enzyme linked immu-nosorbent assay,ELISA)分析两组小鼠外周血和心肌组织炎症因子肿瘤坏死因子-α(tumor necrosisfactor-α,TNF-α)、白细胞介素(interleukin,IL)-1β和IL-6的表达水平.结果观察组小鼠心肌缺血再灌注后,可见心肌组织中炎症细胞大量浸润.与对照组比较,观察组小鼠心肌细胞细胞核中NF-κB水平显著增加,细胞质中p-IκBα水平显著增加,差异具有统计学意义(F值分别为30.497和17.501,P值均<0.05).观察组小鼠外周血中炎症因子TNF-α、IL-1β和IL-6表达水平较对照组显著上调[(373.12±25.19)pg/mL比(178.53±16.01)pg/mL,(919.43±29.88)pg/mL比(119.15±18.09)pg/mL,(519.44±20.14)pg/mL比(136.65±12.41)pg/mL,t值分别为25.250、88.735和62.670,P值均<0.05],心肌组织中炎症因子TNF-α、IL-1β和IL-6表达水平较对照组显著上调[(100.52±10.23)pg/mL比(19.32±4.87)pg/mL,(159.44±12.43)pg/mL比(34.65±8.33)pg/mL,(159.39±14.29)pg/mL比(20.48±8.32)pg/mL,t值分别为27.757、32.300和32.536,P值均<0.05],差异均具有统计学意义.结论心肌缺血再灌注后NF-κB处于激活状态,导致下游炎症因子大量释放,增加了心肌细胞负担.

Objective To investigate the phosphorylation of nuclear factor kappa-B(NF-κB)and the expression of inflammatory factors after myocardial ischemia-reperfusion injury in mice.Methods The model of myocardial ischemia-reperfusion was established with 15 mice by coronary artery ligation and reperfusion as observation group,and another 15 mice were used as control group.Four hours after reperfusion injury,the mice were killed.The pathological changes of myocardial tissue were analyzed by HE staining,and the changes of NF-κB signal pathway were analyzed by Western blot.The expression levels of tumor necrosis factor(TNF-α),interleukin(IL)-1βand IL-6 in peripheral blood and myocardium were analyzed by enzyme-linked immunosorbent assay(ELISA).Results A large number of inflammatory cells infiltrated after myocardial ischemia-reperfusion.Compared with the control group,the level of NF-κB in the nucleus and p-IκBαin the cytoplasm increased significantly(F values were 30.497 and 17.501 respectively,both P values<0.05).The expression level of TNF-α,IL-1βand IL-6 in the peripheral blood of the observation group were also significantly increased than the control group[(373.12±25.19)pg/mL vs(178.53±16.01)pg/mL,(919.43±29.88)pg/mL vs(119.15±18.09)pg/mL,(519.44±20.14)pg/mL vs(136.65±12.41)pg/mL,t values were 25.250,88.735 and 62.670,respectively,all P values<0.05].The inflammatory factor TNF-α,IL-1βand IL-6 in the myocardium were significantly increased in the observation group compared with the control group[(100.52±10.23)pg/mL vs(19.32±4.87)pg/mL,(159.44±12.43)pg/mL vs(34.65±8.33)pg/mL,(159.39±14.29)pg/mL vs(20.48±8.32)pg/mL,t values were 27.757,32.300 and 32.536,respectively,all P values<0.05].Conclusion NF-κB is activated after myocardial ischemia-reperfusion,which leads to the release of downstream inflammatory factors and increases the burden of myocardial cells.