Department of Gastrointestinal, Hernia and Abdominal Wall Surgery, Wuhan Third Hospital (Tongren Hospital of Wuhan University), Hubei Province, 241 Pengliuyang Road, Wuchang District, Wuhan, 430060, China.
Inflamm Res. 2020 Apr;69(4):401-414. doi: 10.1007/s00011-020-01324-2. Epub 2020 Feb 10.
Both innate and adaptive immune system play important roles in the onset and progression of inflammatory bowel diseases (IBDs). However, the significance of natural killer (NK) cells for IBDs remains unclear. To understand the biology of colonic lamina propria natural killer (LPNK) cells in IBDs, we characterized LPNK cell metabolism in a murine acute colitis model.
C57BL/6J mice were fed with 3% dextran sulfate sodium to establish the acute colitis model. Colonic LPNK cells were isolated from mice through flow cytometry. The expression of metabolic genes in LPNK cells was analyzed by transcriptome sequencing and quantitative RT-PCR. Glucose uptake, Seahorse assay, and ATP assay were conducted to assess the metabolic status of LPNK cells. Phos-flow assay was performed to evaluate cell signaling pathways in LPNK cells. In vitro stimulation and cytotoxicity assay were conducted to measure the function of LPNK cells.
In acute colitis, LPNK cells upregulated the expression of genes related to glycolysis and oxidative phosphorylation (oxphos), and enhanced glucose uptake capability. Intracellular ATP production, glycolysis and oxphos in LPNK cells were also promoted in acute colitis. mTORC1 signaling was essential for the metabolic reprogramming in LPNK cells in acute colitis. Although LPNK cells of diseased mice exhibited equivalent cytokine profile to normal LPNK cells upon stimulation with phorbol ester or IL-2, LPNK cells of diseased mice were more cytotoxic to target cells than normal LPNK cells.
LPNK cells undergo metabolic reprogramming which might be a response to upcoming microbial infection in acute colitis.
先天免疫和适应性免疫系统在炎症性肠病（IBD）的发病和进展中都起着重要作用。然而，自然杀伤（NK）细胞在 IBD 中的意义仍不清楚。为了了解 IBD 中结肠固有层自然杀伤（LPNK）细胞的生物学特性，我们在小鼠急性结肠炎模型中对 LPNK 细胞代谢进行了特征描述。
用 3%葡聚糖硫酸钠喂养 C57BL/6J 小鼠建立急性结肠炎模型。通过流式细胞术从小鼠中分离出结肠 LPNK 细胞。通过转录组测序和定量 RT-PCR 分析 LPNK 细胞中代谢基因的表达。进行葡萄糖摄取、 Seahorse 测定和 ATP 测定以评估 LPNK 细胞的代谢状态。进行 Phos-flow 测定以评估 LPNK 细胞中的细胞信号通路。进行体外刺激和细胞毒性测定以测量 LPNK 细胞的功能。
在急性结肠炎中，LPNK 细胞上调了与糖酵解和氧化磷酸化（oxphos）相关的基因表达，并增强了葡萄糖摄取能力。急性结肠炎中 LPNK 细胞的细胞内 ATP 产生、糖酵解和 oxphos 也得到了促进。mTORC1 信号通路对于急性结肠炎中 LPNK 细胞的代谢重编程是必不可少的。尽管与正常 LPNK 细胞相比，受佛波醇酯或 IL-2 刺激后患病小鼠的 LPNK 细胞表现出相当的细胞因子谱，但患病小鼠的 LPNK 细胞对靶细胞的细胞毒性比正常 LPNK 细胞更强。
LPNK 细胞发生代谢重编程，这可能是急性结肠炎中对即将发生的微生物感染的反应。
