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目的:探讨过继移植调节性T细胞(regulatory T cells,Tregs)在小鼠肾脏缺血/再灌注损伤(ischemic/reperfusion injury,IRI)修复中的作用。方法:将24只6周龄C57BL/6小鼠分为假手术组(Sham组)、肾脏IRI组(IR组)、高剂量Tregs移植组(HT组)、低剂量Tregs移植组(LT组)。HT、LT两组提前1 d分别经尾静脉注射5×106、1×105个Tregs。左侧肾蒂夹闭45 min再灌注4 d建立模型,Sham组仅分离左侧肾蒂,不予夹闭。HE染色观察肾脏组织形态学改变;免疫组化和流式检测Tregs浸润情况;免疫组化、Western印迹和(或)ELISA检测Ki67、肿瘤坏死因子(tumor necrosis factor,TNF)-α、白介素(interleukin,IL)-6、IL-10的变化。结果:与IR组和LT组相比,HT组肾组织病理损害减轻,细胞增殖明显升高;Tregs浸润明显增多;IL-10表达增多,TNF-α、IL-6表达减少(P<0.05)。结论:过继移植Tregs可以减轻小鼠缺血再灌注引起的肾脏损伤,且具有剂量依赖性,这可能和其抑制促炎因子TNF-α、IL-6,分泌抗炎因子IL-10有关。
Objective: To investigate the role of adoptive transfer of regulatory T cells (Tregs) in the repair of renal ischemia / reperfusion injury (IRI) in mice. Methods: Twenty-four 6-week-old C57BL / 6 mice were divided into three groups: Sham group, IRI group, HT group, HT group, . The HT and LT groups were injected with 5 × 106 and 1 × 105 Tregs through caudal vein one day earlier. Left renal pedicle clamp 45 min reperfusion 4 d model, Sham group only the left renal pedicle separation, not clipping. Immunohistochemistry and flow cytometry were used to detect the infiltration of Tregs. Immunohistochemistry, Western blotting and / or ELISA were used to detect Ki67, tumor necrosis factor (TNF) -α, interleukin , IL) -6, IL-10. Results: Compared with IR group and LT group, the pathological changes of renal tissue in HT group were alleviated and the cell proliferation was significantly increased; the infiltration of Tregs was significantly increased; the expression of IL-10 increased and the expression of TNF-α and IL-6 decreased (P <0.05) . CONCLUSION: Tregs can reduce renal damage induced by ischemia-reperfusion in a dose-dependent manner, which may be related to the inhibition of proinflammatory cytokines TNF-α, IL-6 and secretion of anti-inflammatory cytokine IL-10.