Changes of renal resident dendritic cells during kidney ischemia-reperfusion injury
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摘要:
目的 探讨在肾脏缺血-再灌注损伤(IRI)期间肾脏固有树突状细胞(rDC)的变化。 方法 采用C57BL/6J小鼠建立双侧肾脏热缺血模型，再灌注24 、48 h后取肾组织制备单细胞悬液，流式细胞仪分析CD45+细胞和CD11c+rDC的比例变化；采用绿色荧光蛋白和白喉毒素受体标记(CD11c+GDTR)的小鼠肾组织制作单细胞悬液，流式细胞仪分析CD11c+rDC的比例及表型；采用CD11c+GDTR小鼠建立双侧肾脏热缺血模型，再灌注24 h后取肾组织制备单细胞悬液，MACS磁珠富集CD45+细胞，经流式细胞仪分析rDC表面共刺激分子表达情况。 结果 再灌注24 h后C57BL/6J小鼠肾脏内CD45+细胞比例明显增加，48 h后其比例进一步升高，再灌注24 h后CD11c+rDC数量同样持续升高，但其占CD45+细胞的比例出现明显下调，48 h后恢复并较Sham组轻度升高；CD11c+GDTR小鼠正常肾脏CD45+细胞比例低于1%，其中约40%为CD11c+的肾脏rDC，主要呈CD11bintF4/80-MHC Ⅱ+；再灌注24 h后CD11c+F4/80-亚群rDC表面共刺激分子CD40、CD80、CD86均显著升高。 结论 热IRI后rDC比例、数量及其表面共刺激分子表达均增加，提示热IRI后肾脏rDC浸润增多且表型成熟。 Abstract:Objective To investigate the changes of renal resident dendritic cells (rDC) during kidney ischemia-reperfusion injury(IRI). Methods C57BL/6J mice models with bilateral renal warm ischemia were established. The kidney tissue was prepared for single cell suspension at 24 h and 48 h after reperfusion. The changes in the percentage of CD45+ cells and CD11c+rDCs were evaluated by flow cytometry. The renal tissues of mice labeled with green fluorescent protein and diphtheria toxin receptor (CD11c+GDTR) were prepared for single cell suspension. The percentage and phenotype of CD11c+rDCs were analyzed by flow cytometry. CD11c+GDTR mice models with bilateral renal warm ischemia were established. The renal tissue was prepared for single cell suspension at 24 h after reperfusion. CD45+ cells was gathered by magnetic-activated cell separation (MACS). The expression levels of co-stimulatory molecules on the rDC surface were analyzed by flow cytometry. Results At 24 h after reperfusion, the percentage of CD45+ cells in the kidney of C57BL/6J mice was significantly elevated, and further increased at 48 h after reperfusion. At 24 h after reperfusion, the quantity of CD11c+rDCs was equally increased, whereas the percentage of CD11c+rDCs in CD45+ cells was dramatically declined and restored at 48 h after reperfusion, slightly higher compared with that in the sham group. In healthy CD11c+GDTR mice, the percentage of CD45+ cells in the kidney was lower than 1%, consisting of approximately 40% of CD11c+rDCs, which mainly presented as CD11bintF4/80-MHCⅡ+. At 24 h after reperfusion, the percentage of CD11c+F4/80- subset rDC surface co-stimulatory molecules was significantly enhanced, such as CD40, CD80 and CD86. Conclusions Following warm IRI, the percentage and quantity of rDCs, and the expression level of rDC surface co-stimulatory molecule are significantly increased, prompting that renal rDC infiltration is increased and phenotype becomes matured. -
Key words:
- Kidney /
- Dendritic cell /
- Ischemia-reperfusion injury /
- Mouse /
- Flow cytometry
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图 1 各组C57BL/6小鼠肾脏rDC比例
注：A图示CD11c+GDTR小鼠CD45+细胞比例；B图示CD11c+GDTR小鼠CD45+细胞中CD11c+的肾脏rDC比例；C图示CD11c+GDTR小鼠肾脏rDC表型；其中Isotype为抗体同型对照，Kidney 1和Kidney 2分别为代表性的两个正常肾脏
Figure 1. Ratios of rDC in kidney of C57BL/6 mice in each group
图 2 CD11c+GDTR小鼠肾脏rDC表型分析
Figure 2. Analysis of renal rDC phenotype in CD11c+GDTR mice
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