脓毒症早期单核细胞减少与预后相关性的分析-现代医学

脓毒症早期单核细胞减少与预后相关性的分析

作者：刘善收¹ 李琳² 代铮¹ 罗许¹ 赵霄君¹ 张锦鑫¹ 梁晓丽¹ 王仙琦¹ 尹文¹

单位：1. 空军军医大学第一附属医院急诊科, 陕西西安 710032;
2. 西安秦皇医院急诊科, 陕西西安 710699

分类号：R459.7

出版年·卷·期（页码）：2023·51·第七期（885-895）

摘要：

目的: 脓毒症早期单核细胞活化并释放炎症因子参与固有免疫反应,炎症过激时免疫细胞大量死亡引发持续免疫抑制,是脓毒症患者死亡的主要原因。本研究探索免疫细胞死亡与预后的相关性,并基于单细胞测序分析脓毒症早期免疫细胞的死亡模式,为优化脓毒症治疗提供依据。方法: 采集脓毒症患者疾病早期临床数据,检测患者血浆内炎症因子、趋化因子、细胞凋亡和焦亡分子的含量。比较不同预后组患者疾病早期临床特征的差异,多因素回归分析患者死亡的危险因素,受试者工作特征(ROC)曲线评估预测效能。扫描电镜和透射电镜观察脓毒症早期免疫细胞形态学改变;通过单细胞测序(sc-RNA seq)揭示不同类型免疫细胞的主要死亡模式。结果: (1) 与存活组相比,死亡患者疾病早期单核细胞计数和百分比均显著降低,患者心率,呼吸频率,血浆ALT、AST、乳酸含量以及SOFA评分显著升高。(2) 多因素回归分析提示单核细胞比例降低、血乳酸增高和入室呼吸次数增多是死亡的危险因素,且单核细胞比例和血乳酸预测效能较好。(3) 脓毒症时血浆炎症因子(TNF-α)、趋化因子(CCL2)、凋亡分子(BAX)、焦亡分子(Caspase1)升高,抗凋亡分子(BCL2L1)显著降低,死亡组更显著;电镜检查提示脓毒症早期免疫细胞呈现典型凋亡和焦亡形态。(4) 单细胞测序揭示脓毒症早期单核细胞内凋亡和焦亡分子显著上调,凋亡和焦亡信号通路显著激活。结论: 本研究发现脓毒症早期单核细胞凋亡和焦亡增加,导致外周单核细胞数量减少;单核细胞比例降低和血浆乳酸增高是脓毒症患者死亡的独立危险因素,两者联合诊断预测效能高,可作为脓毒症早期评估患者预后的方法。

Objective: In the early stage of sepsis, monocytes are activated to release inflammatory factors, participating in the innate immune response. The death of too many immune cells leads to persistent immunosuppression, which is the main cause of death. We explore the correlation between immune cell death and prognosis, and further study on the death pattern of immune cells in early sepsis based on single cell sequencing, so as to provide a basis for optimizing the prevention and treatment of sepsis. Methods: Clinical data of sepsis patients at early stage were collected. The levels of inflammatory cytokines, chemokines, apoptosis and pyroptosis molecules in plasma were detected. The early clinical characteristics of the patients in different prognosis groups were compared. The risk factors of death were analyzed by Logistics/COX regression and the predictive power was evaluated by receiver operating characteristic(ROC) curve. The morphological of immune cells was observed under electron microscopy. Single-cell RNA sequencing revealed a major pattern of death of different types of immune cells in early sepsis, focusing on macrophages and neutrophils involved in the innate immunity. Results: (1) Compared with the survival group, the count and percentage of monocytes in the death group at the early stage of sepsis were significantly lower, but the heart rate and respiratory rate, plasma ALT, AST, and lactate contents, and SOFA score were significantly higher in the death group. (2) The regression analysis showed that the proportion of monocytes, blood lactate and respiratory rate were independent risk factors for prognosis. The ROC curve showed that the proportion of monocytes and the blood lactate value had the best predictive efficacy. (3) The plasma levels of inflammatory factor(TNF-α), chemokine(CCL2), apoptotic molecules(BAX), and pyroptosis molecule(Caspase1) were increased in sepsis, but anti-apoptotic molecule BCL2 was significantly decreased, especially in the death group. Immune cells in early sepsis showed typical apoptotic and pyrolytic morphology. (4) Single cell sequencing showed that apoptotic and pyroptosis molecules were significantly up-regulated in monocytes at the early stage of sepsis, and apoptosis and pyroptosis signal pathways were significantly activated. Conclusion: This study found that in the early stage of sepsis, the degree of monocyte apoptosis and pyroptosis increased significantly, resulting in a decrease in the number of peripheral monocytes. Reduced monocyte ratio and increased plasma lactic acid are risk factors for death due to sepsis. The combined diagnosis of the two indicators has high predictive power and can be used as a method for early evaluation of the prognosis of patients with sepsis.

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