巨噬细胞内IKKα在小鼠肾脏缺血再灌注损伤炎症反应中的作用
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国家自然科学基金(81370797)


The role of IKKα in macrophages in renal inflammation after renal ischemia-reperfusion injury in mice
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    摘要:

    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。

    Abstract:

    Objective:To study the role of IKKα in macrophages in renal inflammation after renal ischemia-reperfusion injury(IRI)in mice model.Methods:A total of 20 C57BL/6 wild type (WT) mice and 20 IKKαMKO (KO) mice (healthy,male,and aged 8~10 weeks) were randomly divided into the control (Sham) group and the renal IRI group,respectively.The morphology change of renal tissue and the infiltration of inflammatory cells were observed by hematoxylin and eosin (HE) staining.The expressions of interleukin 10(IL-10),interleukin 6(IL-6),Ki67,CD68,iNOS,and Arg-1 were detected by immunohistochemistry.The protein expressions of IL-10 and IL-6 were detected by Western blotting.Results:Compared with the Sham group,obvious renal tissue injury and increasing inflammation infiltration were observed in the IRI group by HE staining. Immunohistochemical results on expressions of both IL-10 and IL-6 were increased after renal IRI occurred,IL-10 increased while IL-6 decreased by time.Compared with the WT-IRI group,the renal pathological damage was more severe (P<0.01),the expressions of IL-6,CD68 and iNOS were significantly increased (P<0.01),but the expressions of IL-10 (P<0.01) and Ki67 (P<0.05) were distinctly decreased in the KO-IRI group.Conclusion:IKKα knockout in macrophages aggravates renal ischemia-reperfusion injury and inhibits renal repair in mice model,which can be related to intensifying the early infiltration and inflammatory reaction of macrophages(M1 mainly).

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王 晶,万 辛,黄文娟,陈 文,曹长春.巨噬细胞内IKKα在小鼠肾脏缺血再灌注损伤炎症反应中的作用[J].南京医科大学学报(自然科学版),2017,(4):403-408

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  • 收稿日期:2016-11-09
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  • 在线发布日期: 2017-05-10
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