摘要
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 study the effect of adoptive transfer of regulatory T cells（Tregs）on the repair phase of ischemia / reperfusion（IR）induced renal injury in mice. Methods：A total of 24 C57BL / 6 mice aged 6-week-old were divided into Sham group, IR group,high-dose transfer group（HT）,and low-dose transfer group（LT）. Approximately 5 × 106and 1 × 105Tregs were injected intravenously into mice by tail vein 1 d before IR injury induced. Left renal pedicle was bluntly dissected and clamped with a microvascular clamp for 45 min and reperfusion 4 d to establish IR renal injury model. Left renal pedicle was only separated without clipping in the Sham group. HE staining was performed to evaluate morphology changes of renal tissues. Immunohistochemical and flow cytometry were performed to detect the infiltrating of Tregs（CD4＋CD25＋）;the expression changes of Ki67,tumor necrosis factor-α（TNF-α）,IL-16 and IL-10 were determined by immunohistochemistry,Western blotting and / or ELISA. Results：Compared to the IR group and the LT group,the damage of renal pathological was reduced,cell proliferation,the infiltration of Tregs and the expression of IL-10 were significantly increased（P ＆lt; 0.05）,and the expression of TNF-α and IL-6 were decreased（P ＆lt; 0.05）in the HT group. Conclusion：Adoptive transfer of Tregs had a dose-dependent protective effect on renal ischemia-reperfusion-induced injury,which is possibly related to inhibition of the proinflammatory cytokines TNF-α,IL-6 and increase of anti-inflammatory cytokine IL-10.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2014年第3期303-309,共7页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金(81170658)
关键词
调节性T细胞
肾脏
缺血再灌注损伤
修复
regulatory T cells
kidney
ischemia reperfusion injury
repair
