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目的:研究巨噬细胞内IKKα在小鼠肾脏缺血再灌注损伤(ischemic reperfusion injury,IRI)炎症反应中的作用。方法:分别将20只8~10周龄健康雄性C57BL/6小鼠(WT小鼠)、20只8~10周龄健康雄性巨噬细胞内IKKα基因敲除即IKKαMKO小鼠(KO小鼠)随机分为假手术(Sham)组和肾脏缺血再灌注损伤(IRI)组,分别构建模型。苏木素-伊红(HE)染色法观察肾脏组织形态学改变及炎症细胞浸润情况,免疫组织化学法检测肾组织抑炎因子白介素-10(IL-10)、促炎因子白介素-6(IL-6)、增殖指标Ki67、巨噬细胞标记物CD68、M1型巨噬细胞标记物诱导型一氧化氮合酶(iNOS)、M2型巨噬细胞标记物精氨酸酶1(Arg-1)的表达,Western blot检测IL-10、IL-6的表达变化。结果:HE染色结果表明IRI组较Sham组肾组织结构损伤明显及炎症浸润增加。免疫组化结果表明肾脏IRI后IL-10和IL-6均呈高表达,IL-10表达随时间延长而递增,IL-6表达随时间延长而递减。与WT-RI组相比,KO-IRI组小鼠肾脏病理损伤加重(P<0.01),IL-6、CD68、iNOS表达显著增加(P<0.01),IL-10(P<0.01)、Ki67(P<0.05)表达显著降低。结论:巨噬细胞内IKKα基因敲除加重了小鼠肾脏缺血再灌注损伤的炎症反应且不利于肾脏修复,这可能与增加了肾脏早期巨噬细胞(M1型为主)浸润及促进了炎症反应有关。
Objective: To investigate the role of IKKα in macrophage inflammatory response in mice with renal ischemia-reperfusion injury (IRI). Methods: IKKα gene knockout (IKKαMKO) mice (KO mice) were cultured in 20 healthy male C57BL / 6 mice (WT mice) aged 8-10 weeks and 20 healthy male macrophages aged 8-10 weeks The rats were randomly divided into Sham group and IRI group. The morphological changes of renal tissues and the infiltration of inflammatory cells were observed by hematoxylin and eosin (HE) staining. The expressions of interleukin-10 (IL-10), interleukin-6 ), Proliferation index Ki67, macrophage marker CD68, M1-induced inducible nitric oxide synthase (iNOS), and M2-marker arginase 1 (Arg-1) , Western blot detection of IL-10, IL-6 expression changes. Results: The results of HE staining showed that the injury of renal tissue and the infiltration of inflammation in IRI group were more obvious than those in Sham group. The results of immunohistochemistry showed that the expression of IL-10 and IL-6 in kidneys were high after IRI, the expression of IL-10 increased with time and the expression of IL-6 decreased with time. Compared with WT-RI group, the pathological changes of kidney in KO-IRI group were significantly increased (P <0.01), and the expressions of IL-6, CD68 and iNOS were significantly increased (P <0.01) (P <0.05) expression was significantly reduced. CONCLUSION: IKKα knockout in macrophages aggravates inflammatory reaction of renal ischemia-reperfusion injury in mice and is not conducive to renal repair, which may be associated with increased infiltration of early renal macrophages (type M1 predominance) and inflammation Reaction related.