目的 通过分析新型冠状病毒感染后（简称“新冠后”）慢性疲劳综合征（chronic fatigue syndrome，CFS）患者与正常受试者的单细胞转录组数据，探讨新冠后CFS与炎症反应、氧化应激和细胞凋亡之间的分子机制，并预测调控关键基因的潜在中药，为治疗新冠后CFS提供理论支持。方法 使用R语言4.2.1中的“Seurat”包处理新冠后CFS患者与正常受试者的外周血单细胞测序数据，“CellChat”包进行细胞通讯分析，“scMetabolism”包评估细胞代谢特征，综合5种算法（AddModuleScore、AUCell、UCell、singscore和ssGSEA）计算新冠后CFS患者与正常受试者中5种免疫细胞炎症反应、氧化应激和细胞凋亡评分，并比较两者间的差异。使用“scCODE”包鉴定各类免疫细胞在正常受试者与新冠后CFS患者之间的差异表达（differential expression，DE）基因。Upset图分析得到新冠后CFS炎症反应相关基因（inflammatory response related genes，IRGs）、氧化应激相关基因（oxidative stress related genes，OSRGs）和细胞凋亡相关基因（apoptosis related genes，ARGs）。使用“ClusterProfiler”包分别对IRGs、OSRGs、ARGs进行基因本体生物学过程（gene ontology biological process，GOBP）富集分析。基于STRING数据库及Cytoscape软件构建“IRGs-OSRGs-ARGs”分子交互复杂网络，使用cytoHubba插件筛选关键基因。利用COREMINE数据库预测调控关键基因的中药，并通过古今医案云平台统计中药的性味归经、药物功效分类。结果 共鉴定出5种免疫细胞，分别为B细胞、CD4⁺ T细胞、CD8⁺ T细胞、NK细胞和单核细胞。相比于正常受试者，新冠后CFS患者的细胞通讯更为丰富且强度更大，两者之间并无明显差异的细胞代谢特征。新冠后CFS患者中各类免疫细胞的炎症反应、氧化应激和细胞凋亡评分普遍高于正常受试者。共得到33个IRGs、17个OSRGs和54个ARGs。“IRGs-OSRGs-ARGs”分子交互网络显示，IRGs、OSRGs和ARGs之间存在密切联系，最后鉴定出8个关键基因。干预中药药性以寒为主，温、平次之；药味以苦、辛、甘为主；归经以肝、脾、肺为主；功效分类以补虚药（甘草、人参等）、清热药（黄芩、鱼腥草等）、活血化瘀药（郁金、丹参等）为主。结论 新冠后CFS患者存在异常的炎症反应、氧化应激和细胞凋亡状态，且三者之间密切关联。8个关键基因（TNF、BCL2、CTNNB1、CXCL8、IFNG、HIF1A、MCL1、CYCS）可能是新冠后CFS的潜在治疗靶标。临床治疗新冠后CFS应注重补虚药、清热药及活血化瘀等药物的配伍使用。

Objective To explore the molecular mechanism between chronic fatigue syndrome (CFS) after coronavirus disease 2019 (COVID-19) and inflammatory response, oxidative stress, apoptosis by analyzing the single cell transcriptome data of patients with CFS after COVID-19 and normal subjects, and predict potential traditional Chinese medicines that regulate key genes, providing theoretical support for the treatment of CFS after COVID-19. Methods The “Seurat” package in R language 4.2.1 was used to process peripheral blood single cell sequencing data of CFS after COVID-19 patients and normal subjects; the “CellChat” package was used to analyze cell communication; and the “scMetabolism” package was used to evaluate cell metabolic characteristics. Five algorithms (AddModuleScore, AUCell, UCell, singscore and ssGSEA) were synthesized to calculate the inflammatory response, oxidative stress, apoptosis scores of five immune cells in CFS after COVID-19 patients and normal subjects, and the differences between them were compared. The “scCODE” package was used to identify differentially expressed (DE) genes between normal subjects and CFS after COVID-19 patients in various types of immune cells. Upset plot analysis showed that inflammatory response related genes (IRGs), oxidative stress related genes (OSRGs) and apoptosis related genes (ARGs) were involved in CFS after COVID-19. “ClusterProfiler” package was used to perform gene ontology biological process (GOBP) enrichment analysis on IRGs, OSRGs and ARGs respectively. STRING database and Cytoscape software were used to construct “IRGs-OSRGs-ARGs” molecular interaction complex network diagram. The cytoHubba plug-in was used to screen for key genes. COREMINE database was used to predict traditional Chinese medicine regulating key genes, and qi, flavours, meridian affinity, and drug efficacy classification were counted through the cloud platform of ancient and modern medical records. Results Five types of immune cells were identified, including B cells, CD4⁺T cells, CD8⁺T cells, NK cells, and monocytes. Cell-to-cell communication was richer and more intense in CFS after COVID-19 patients than in normal subjects, and there were no significant differences in cellular metabolic characteristics between the two groups. The scores of inflammatory response, oxidative stress, apoptosis of immune cells in CFS after COVID-19 patients were generally higher than those in normal subjects. A total of 33 IRGs, 17 OSRGs and 54 ARGs were obtained. “IRGs-OSRGs-ARGs” molecular interaction network showed that IRGs, OSRGs and ARGs were closely related, and finally eight key genes were identified. The qi of the intervention traditional Chinese medicines is primarily cold, followed by warm and neutral. The flavors were mainly bitter, pungent and sweet. Liver, spleen, and lung channels were the main channels. The main efficacy categories were deficiency tonifying drugs [Gancao (Glycyrrhizae Radix et Rhizoma), Renshen (Ginseng Radix et Rhizoma), etc.], heat-clearing drugs [Huangqin (Scutellariae Radix), Yuxingcao (Houttuyniae Herba), etc.], blood-activating and stasis- removing drugs [Yujin (Curcumae Radix), Danshen (Salviae Miltiorrhizae Radix et Rhizoma), etc.]. Conclusion CFS after COVID-19 patients had abnormal inflammatory response, oxidative stress, apoptosis and the three were closely related. A total of eight key genes (TNF, BCL2, CTNNB1, CXCL8, IFNG, HIF1A, MCL1, CYCS) may be potential therapeutic targets for CFS after COVID-19. In clinical treatment of CFS after COVID-19, attention should be paid to the compatibility of tonifying drugs, heat-clearing drugs and blood-activating and stasis-removing drugs.

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国家中医药管理局中医药创新团队及人才支持计划项目（ZYYCXTD-D-202203）；国家中医药管理局项目（2023ZYLCYJ02-21）；广东省重点领域研发计划项目（2023B1111020003）；广东省中医院重点实验室专项（JZ2020ZZ01，YN2023JZ02，KF2023JZ06）；广州市科技局市校院联合资助项目—广州市中西医结合防治新发突发传染病重点实验室（202201020382）；中华中医药学会临床研究项目（2023DEPLHGG-06）；广东省中医院朝阳人才科研专项资助（ZY2022KY10，ZY2022YL04）