巨噬细胞在小鼠肾脏缺血/再灌注损伤修复中的作用

Role of macrophage on the repair phase of ischemic/reperfusion renal injury in mice

目的 探讨巨噬细胞在小鼠肾脏缺血/再灌注（IR）损伤修复期中的作用.方法 24只6～8周龄C57BL/6小鼠被分为假手术（Sham）组、缺血再灌注（IR）组、氯磷酸盐脂质体（LC）组、空白对照（Ctl）组.Sham组仅切开背部皮肤,暴露双侧肾蒂不予夹闭;其余3组小鼠均给予夹闭双侧肾蒂25 min后恢复血流,再灌注3d后留取肾脏标本.LC组和Ctl组在留取肾脏标本前1天分别经腹腔注射LC（0.15～ 0.2 ml/20 g）或PBS.HE染色观察肾脏组织形态学改变;免疫组化法检测巨噬细胞浸润情况,Ki67蛋白、肿瘤坏死因子（TNF-α）和白细胞介素（IL）10的表达;Western印迹法检测TNF-α和IL-10的改变.结果 与IR组和Ctl组相比,LC组肾组织巨噬细胞浸润明显减少,肾组织病理损伤加重,细胞增殖减少,TNF-α表达明显增多,IL-10表达明显减少（均P＜ 0.05）.结论 在IR损伤修复期敲除巨噬细胞可加重小鼠缺血再灌注损伤,可能和巨噬细胞可抑制促炎因子TNF-α表达和促抗炎因子IL-10分泌有关.

Objective To study the effect of macrophage on the repair phase of ischemic/ reperfusion （IR） renal injury in mice.Methods A total of 24 C57BL/6 mice aged 6 ～ 8 week-old were divided into 4 groups：the Sham group,IR group,LC group and Control group.For all the groups,the bilateral renal pedicles of the mice were clipped after dorsal skin dissection,except for the Sham group,then unclipped them 25 minutes later to restore blood flow to the kidney and collected the renal specimens after 3 days.The LC group and Control group each were injected intraperitoneally with 0.15 ～ 0.20 ml/20 g per day before kidney specimens were taken.The morphology changes of renal tissues were evaluated by HE staining.Immunohistochemical testing was performed to detect the infiltration of macrophages,the expression change of Ki67,TNF-α and IL-10.In addition TNF-o,IL 10 were measured using Western blotting.Results Compared to the IR group and the Control group,the infiltration of macrophages was markedly decreased,the damage of renal pathology was aggravated,the cell proliferation was significantly decreased,and the expression of IL-10 was also decreased （P ＜ 0.05）,while the expression of TNF-α were increased （P ＜ 0.05）.Conclusions Intraperitoneal LC injection can aggravate kidney damage on the repair phase of renal ischemic/reperfusion injury in mice,which is possibly related its inhibition of proinflammatory cytokines TNF-α and the secretion of antiinflammatory cytokine IL-10.