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外周血自然杀伤细胞CX3CR1在脓毒症患者中的表达特征及其与肠道菌群的关系

Characteristics of the expression of CX3CR1 in natural killer cells from peripheral blood and its association with gut microbiota in sepsis patients

    摘要
  • 摘要:
    目的 探讨脓毒症患者外周血单个核细胞（peripheral blood mononuclear cell, PBMC）中自然杀伤（natural killer, NK）细胞CX3CR1表达量的变化规律及其与肠道菌群的关系。
    方法 选择2020年1月至2021年1月在复旦大学附属中山医院诊治的24例脓毒症患者，另于2021年1月招募10名健康志愿者作为健康对照。收集脓毒症患者入院第1天和第4天的粪便及外周血。通过Illumina MiSeq平台对肠道菌群的16S rDNA基因V3～4进行测序。使用CD56阳性磁珠分选，并以流式细胞术鉴定外周血CD3－CD56＋NK细胞，采用qPCR检测CX3CR1表达量。分析NK细胞中CX3CR1表达量变化以及其与肠道菌群的相关性。
    结果 与健康对照组相比，脓毒症患者肠道菌群Shannon多样性指数及厚壁菌门（Firmicutes）占比降低；与入院第1天相比，脓毒症患者入院第4天肠道菌群Shannon多样性指数减小，变形菌门（Proteobacteria）占比升高，肠球菌属（Enterococcus）与克雷伯菌属（Klebsiella）相对含量增加。脓毒症患者入院第4天外周血NK细胞的CX3CR1表达量低于入院第1天（P＜0.001）；与存活患者相比，死亡患者入院第4天的CX3CR1表达量降低（P＜0.05）。Spearman相关分析显示，外周血NK细胞CX3CR1表达量与肠道菌群数量及Shannon多样性指数正相关（P＜0.01）。
    结论 脓毒症患者外周血NK细胞CX3CR1表达量随病程进展下降，并与预后相关，其表达与肠道菌群密切相关。

     

    Abstract:
    Objective To explore the changes in the expression of CX3CR1 in natural killer (NK) cells from peripheral blood mononuclear cells (PBMC) in sepsis patients and its association with gut microbiota.
    Methods A total of 24 sepsis patients were selected from January 2020 to January 2021 at Zhongshan Hospital, Fudan University, and 10 healthy volunteers were recruited in January 2021 as healthy controls. Fecal samples and peripheral blood were collected from sepsis patients on the first and fourth days of hospitalization. Sequencing of the V3-4 region of the 16S rDNA gene of gut microbiota was performed using the Illumina MiSeq platform. The peripheral blood samples were isolated by positively selected magnetic beads, and the CD3－CD56＋NK cells were identified by flow cytometry. The mRNA expression of CX3CR1 was detected by qPCR, and the changes in CX3CR1 expression and its correlation with gut microbiota were analyzed.
    Results Compared with healthy control group, the Shannon diversity index of the gut microbiota and the proportion of Firmicutes in sepsis patients decreased; compared with admission day, the Shannon diversity of the gut microbiota in sepsis patients on the fourth day of hospitalization significantly decreased, the proportion of Proteobacteria on phylum level and the relative abundance of Enterococcus and Klebsiella on genus level significantly increased. The CX3CR1 expression of PBMC-NK cells in sepsis patients on the fourth day was significantly lower than that on the admission day (P＜0.001). Compared with surviving patients, CX3CR1 expression in non-surviving patients significantly decreased on the fourth day (P＜0.05). Spearman correlation analysis showed that CX3CR1 expression of PBMC-NK cells was positively correlated with the quantity of gut microbiota and the Shannon diversity index (P＜0.01).
    Conclusions The expression of CX3CR1 in PBMC-NK cells in sepsis patients decreases with disease progression, and is related to prognosis. Furthermore, its expression is found to be closely related to the gut microbiota.

     

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