IKKα对缺血再灌注损伤小鼠的保护作用及其对M1与M2型巨噬细胞的影响  

Protective Effect of IKKɑon Mice with Ischemia-reperfusion Injury and Its Effects on M1 and M2 Macrophages

作　　者：陈波 张金盈[2] 马平[2] CHEN Bo;ZHANG Jinying;MA Ping(Zhengzhou Central Hospital,Zhengzhou 475000,Henan,China;First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,Henan,China)

机构地区：[1]郑州市中心医院,郑州475000 [2]郑州大学第一附属医院,郑州450000

出　　处：《中西医结合心脑血管病杂志》2020年第11期1707-1712,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学基金面上项目(No.81570274)。

摘　　要：目的探讨IκB激酶复合物(IKKα)对缺血再灌注(IR)损伤小鼠的保护作用及其对M1与M2型巨噬细胞的影响。方法C57BL/6-IKKα条件性基因敲除小鼠(mIKKα-/-)及同窝出生的年龄性别匹配的对照组小鼠各9只,各自分为MIRI模型组(6只)和假手术组(3只),以上各组小鼠分别于IR3 d及7 d后进行心脏超声及血流动力学检测;IR模型组于IR3 d及7 d后分别处死(3只),HE及Masson染色进行心肌病理学观察,免疫组织化学法检测心肌组织中炎性因子白细胞介素-12A(IL-12A)、肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)、CD68相对表达量,Western Blot法检测心肌组织中M1/M2型巨噬细胞表型标记物诱导型一氧化氮合酶(iNOS)、表型标记物(Arg1)蛋白相对表达量,实时荧光定量PCR法检测心肌组织中M1型巨噬细胞表型标记物(iNOS、TNF-α、IL-1β)mRNA的表达量以及促M2巨噬细胞因子[肿瘤坏死因子-β(TGF-β)、白细胞介素-10(IL-10)]、M2巨噬细胞Arg1mRNA的表达量。结果与IKKαflox/flox模型组比较,mIKKα-/-模型组IR 3 d及7 d的左心室舒张末期内径(LVIDd)、心肌梗死面积及胶原附着面积、炎性因子IL-12A、IL-10、CD68阳性区域评分、M1型巨噬细胞表型标记物iNOS蛋白量及iNOS、TNF-α和IL-1βRNA的表达量均增加(P<0.05);左室射血分数(EF)、左室室壁心肌纤维缩短率(FS)、M2型巨噬细胞表型标记物Arg1蛋白量和Arg1mRNA的表达量均降低(P<0.05);促M2巨噬细胞极化因子(TGF-β、IL-10)mRNA表达量比较差异无统计学意义(P>0.05)。结论IKKα对缺血再灌注损伤小鼠的炎症反应具有保护作用,该作用可能与IKKα调节心肌巨噬细胞聚集并向M1型巨噬细胞活化有关。Objective To investigate the protective effect of IκB kinaseɑ(IKKɑ)on ischemia-reperfusion injury(IR)mice and its effects on M1 and M2 macrophages.Methods C57BL/6-IKKɑconditional gene knockout mice(mIKKɑ-/-)and control group-aged sex-matched control mice were each divided into IR model group(n=6)and sham operation group(n=3).The mice in the above groups were examined by echocardiography and hemodynamics after 3 d and 7d of IR,respectively.The mice in the IR model group(n=3)were sacrificed after 3 d and 7d of IR.Hematoxylin-Eosin(HE)staining and Masson staining were used to observe myocardial pathology.Immunohistochemistry was used to detect the inflammatory factors interleukin(IL)-12A,tumor necrosis factor-α(TNF-α),and IL-10,CD68 relative expressions in myocardial tissue.Western blot was used to detect the relative expression of M1/M2 macrophage phenotype markers iNOS and Arg1 protein in myocardial tissue.Real-time fluorescence quantitative polymerase chain reaction(PCR)was used to detect the expression of M1 macrophage phenotype markers(iNOS,TNF-α,IL-1β)mRNA expression,M2 macrophage-stimulating factor[tumor necrosis factor-β(TGF-β),IL-10],M2 macrophage phenotype marker(Arg1)mRNA expression in myocardial tissue.Results Compared with the IKKαflox/flox model group,the left ventricular end-diastolic diameter,myocardial infarct size and collagen attachment area,inflammatory factor IL-12A,IL-10,CD68 positive region score,M1 macrophage in mIKKɑ-/-model group,the amount of iNOS protein,and the expressions of iNOS,TNF-αand IL-1βmRNA were significantly increased after 3 d and 7d of IR(P<0.05).Left ventricular ejection fraction and fractional shortening,M2 type macrophage phenotype markers Arg1 protein and Arg1 mRNA expression levels were reduced(P<0.05).There was no statistically significant difference in the expression of M2 macrophage polarization factor(TGF-β,IL-10)mRNA(P>0.05).Conclusion IKKɑhas a significant protective effect on inflammatory response in mice with IR injury,which may be related to IKK

关 键 词：心肌缺血再灌注 巨噬细胞 IκB激酶复合物 心肌梗死面积 炎性因子 

分 类 号：R542.2[医药卫生—心血管疾病] R285.5[医药卫生—内科学]