第一章 心脏骤停后的临床管理现状与炎症控制需求分析

研究背景：心脏骤停后脑损伤的病理机制涉及缺血再灌注损伤诱导的炎症级联反应与免疫代谢紊乱，而现有心脏骤停后的临床管理对复苏后炎症反应控制及脑保护策略的不足可能加剧患者的神经功能损伤。

研究目的：通过评估我国心脏骤停后临床管理现状，寻找心骤停药物治疗的研究成果向临床实践转化面临的困难与挑战，揭示心脏骤停后炎症控制与脑保护的实践缺口，为未来靶向干预策略的临床转化提供依据。

研究方法：开展全国范围的线上问卷调查（56项问题），涵盖医疗机构基本特征、心脏骤停处置流程与复苏质量监测、胸外按压与通气、药物应用、高级生命支持及复苏技能培训等维度（如ECPR、目标体温管理）及质量监测等维度，分析标准化实践差异对炎症控制的影响。

研究成果与结论：共回收有效问卷814份（涉及23个省、4个自治区及4个直辖市）。问卷结果显示我国心脏骤停后临床管理呈现出“指南框架下的执行差异性”，具体表现为初始按压以人工为主，器械按压在后续复苏中使用率逐步提升；约半数施救者首选球囊面罩通气而非建立高级气道，自主循环恢复后75%的病例转为机械通气；气管插管是高级气道建立的主要方式；14%的施救者在复苏过程中会使用糖皮质激素控制炎症；按压反馈装置在ROSC评估中作用日益凸显；区域性医疗中心ECPR与目标体温管理技术应用能力不足且实践标准尚未统一。这些管理流程中的非标准化可能助推心脏骤停后缺血再灌注损伤的炎症级联反应过度激活，并可能会阻碍靶向神经保护策略的临床转化，也因此提示亟需探索靶向调控炎症级联反应的干预手段。

第二章 JAK3 抑制剂对心脏骤停后脑组织缺血再灌注损伤的保护作用

研究目的：第一章揭示了对于心脏骤停后炎症控制与脑保护的需求，本章进一步通过动物模型探究JAK3-STAT5通路在心脏骤停后脑损伤中的激活情况及JAK3特异性抑制剂（JANEX-1）靶向抑制JAK3 活化对心脏骤停后脑损伤的保护作用。

研究内容：构建大鼠心脏骤停-心肺复苏模型，分析心脏骤停后脑组织中JAK3-STAT5信号通路激活情况，并通过JANEX-1抑制JAK3磷酸化，阻断信号转导，观察其对心脏骤停早期炎症指标、脑损伤标志物、T淋巴细胞浸润、细胞凋亡等病理过程的影响。

研究方法：大鼠随机分为Sham组、CPR-Only组、CPR-Instant组、ROSC-10min组与DMSO溶剂对照组，构建心脏骤停-心肺复苏模型。在取得ROSC 2h及24h采集外周血以及ROSC 24h脑组织，检测外周血中IL-6、10、TNF-α、IFN-γ及CRP等炎症指标；NSE、GFAP、S100β、Nf-L、UCH-L1、总Tau等脑损伤标志物；大鼠脑组织海马与皮质的JAK3/p-JAK3、STAT5/p-STAT5以及c-Caspase3、Bcl-2等凋亡蛋白。对ROSC 24h的脑组织进行HE染色及TUNEL染色评估脑组织在JANEX-1干预前后的病理改变及细胞凋亡情况。免疫荧光法检测CD4⁺和CD8⁺T淋巴细胞浸润。

研究成果和结论：本研究发现心脏骤停后脑组织（尤其海马区）的JAK3-STAT5信号通路被显著激活。JANEX-1可对心脏骤停后脑损伤发挥抗炎、抗凋亡、免疫调节等多重神经保护作用，并在心脏骤停早期表现出潜在时间窗口依赖性及作用区域差异性。这些发现为理解心脏骤停后脑损伤相关的多靶点协同免疫调控网络提供了理论依据，并提示JAK3-STAT5通路可作为脑损伤的潜在治疗靶点。

第三章 JAK3—STAT5通路相关基因在心脏骤停患者中的表达情况

研究目的：前两章分别从临床实践缺口和动物模型层面论证了JAK3-STAT5通路干预的必要性，本章进一步通过临床队列分析心脏骤停患者JAK3-STAT5通路相关基因的表达特征及其与炎症、免疫代谢紊乱的关联。

研究内容：本研究是在2024年6月~2025年1月之间开展的单中心病例对照研究，比较了心脏骤停患者与非心脏骤停患者起病早期外周血JAK3-STAT5通路相关基因表达差异及通路基因与脑损伤标志物、炎症、代谢指标等的相关性。

研究方法：本研究纳入我院抢救室接收的非外伤性院前或院内心脏骤停患者，对照组为同期的其他非心脏骤停患者。对符合纳入标准的患者采集外周静脉血，用于分析JAK3-STAT5通路及相关转录因子的基因，包括JAK3，STAT5a、STAT5b、FoxP3、T-bet、GATA3、RORγt、HIF-1α，以及NSE、GFAP、S100β、Nf-L、总Tau、UCH-L1等脑损伤标志物。主要结局指标为心脏骤停组与非心脏骤停组的JAK3-STAT5通路及相关转录因子的基因表达差异；次要结局主要包括以上两组患者脑损伤标志物及ROSC组与非ROSC组JAK3-STAT5相关通路基因及脑损伤标志物的差异。

研究成果和结论：心脏骤停患者与非心脏骤停患者外周血中JAK3、STAT5a、STAT5b的基因表达未见明显差异；但心脏骤停组HIF-1α和GATA3基因表达量显著升高，ROSC组中GATA3和T-bet表达降低；而且JAK3-STAT5通路基因表达与心脏骤停患者糖尿病史、PaO₂和血浆白蛋白水平存在相关性，提示该通路可能通过协调Th细胞分化与参与代谢稳态影响复苏过程。为理解心脏骤停后免疫代谢紊乱提供了新视角，有望为个体化免疫治疗提供潜在靶点。

Chapter 1: Current Status of Post-Cardiac Arrest Clinical Management and Analysis of Inflammation Control Needs
Background: The pathological mechanisms of post-cardiac arrest brain injury involve inflammatory cascades and immunometabolic dysregulation induced by ischemia-reperfusion injury. However, deficiencies in current clinical management strategies for post-resuscitation inflammation control and neuroprotection may exacerbate neurological damage.
Objective: To evaluate the current status of post-cardiac arrest clinical management in China, identify challenges in translating pharmacological research into clinical practice, and reveal gaps in inflammation control and neuroprotection, thereby providing a foundation for future targeted interventions.
Methods: A nationwide online questionnaire (56 items) was conducted, covering institutional characteristics, cardiac arrest management protocols, resuscitation quality monitoring (e.g., chest compression, ventilation, drug administration), advanced life support (e.g., ECPR, targeted temperature management), and training. The impact of standardized practice variations on inflammation control was analyzed.
Results and Conclusions: A total of 814 valid questionnaires were collected (spanning 23 provinces, 4 autonomous regions, and 4 municipalities). Results revealed "guideline-driven implementation heterogeneity": initial manual compressions dominated (71%), with gradual uptake of mechanical devices; 49% preferred bag-mask ventilation initially, while 75% transitioned to mechanical ventilation after ROSC; endotracheal intubation was the primary advanced airway method; 14% used glucocorticoids for inflammation control; feedback devices were increasingly utilized for ROSC assessment. Regional disparities in ECPR and targeted temperature management protocols were noted. These variations may exacerbate ischemia-reperfusion-induced inflammatory cascades and hinder the translation of targeted neuroprotective strategies, underscoring the urgent need for precision interventions.

Chapter 2. The Protective Effect of JAK3 Inhibitor on Cerebral Ischemia-Reperfusion Injury After Cardiac Arrest

Objective：Building on the identified clinical needs for inflammation control and neuroprotection (Chapter 1), this study investigates investigate the activation of JAK3-STAT5 pathway in post-cardiac arrest brain injury and evaluate the therapeutic potential and neuroprotective effects of JAK3-specific inhibitor, JANEX-1 by targeting JAK3 activation.

Research Scope: A rat model of cardiac arrest was established to analyze the activation dynamics of JAK3-STAT5 signaling in post-cardiac arrest brain injury. JANEX-1 was administered to inhibit JAK3 phosphorylation, thereby blocking downstream signal transduction. We evaluated the effects of JAK3 inhibition on early inflammatory markers (e.g., IL-6,IL-10,TNF-α,IFN-γ,CRP), brain injury biomarkers (e.g., NSE, GFAP, S100β,Nf-L,UCH-L1,Tau), T-lymphocyte infiltration (CD4⁺/CD8⁺), apoptosis, and histopathological changes in cerebral tissues after CA.

Methods: Rats were randomized into five groups (Sham, CPR-Only, CPR-Instant, ROSC-10min, and DMSO vehicle control. Peripheral blood and brain tissues (hippocampus and cortex) were collected at 2h and 24h post-return of spontaneous circulation. Inflammatory cytokines, brain injury markers, JAK3-STAT5 pathway related proteins (JAK3, p-JAK3, STAT5, p-STAT5), apoptosis related proteins were quantified. Histopathological and apoptotic assessments were performed using hematoxylin-eosin (HE) and TUNEL staining. Immunofluorescence was employed to evaluate T-cell infiltration.

Results and Conclusions: This study demonstrates that the JAK3-STAT5 signaling pathway was significantly activated in post-cardiac arrest brain injury, particularly in the hippocampal region. JANEX-1 exhibited multi-target neuroprotective effects, including anti-inflammatory, anti-apoptotic, and immunomodulatory properties, with superior efficacy when administered early. Time-dependent and region-specific therapeutic effects were observed, highlighting its potential for clinical translation. These results provide a theoretical foundation for understanding the synergistic immune regulatory network in post-cardiac arrest brain injury and propose JAK3-STAT5 as a promising therapeutic target.

Chapter 3. Expression of JAK3-STAT5 Pathway Related Genes in Patients after Cardiac Arrest

Objectives：Complementing prior clinical and preclinical evidence (Chapters 1–2), this study analyzes the expression characteristics of JAK3-STAT5 pathway-related genes in CA patients and their association with inflammation and immune-metabolic dysregulation.

Research Scope：This single-center case-control study, conducted from June 2024 to January 2025, compares early peripheral blood gene expression profiles of the JAK3-STAT5 pathway and its associated transcription factors between cardiac arrest (CA) patients and non-CA controls, and correlations between pathway genes and brain injury biomarkers, inflammatory markers, and metabolic indicators.

Methods：Non-traumatic out-of-hospital or in-hospital CA patients admitted to our emergency department were enrolled, with non-CA patients serving as controls. Peripheral blood samples were collected to analyze brain injury biomarkers (NSE, GFAP, S100β, Nf-L, total Tau, UCH-L1) and differential gene expression in the JAK3-STAT5 pathway and related transcription factors (JAK3, STAT5a, STAT5b, FoxP3, T-bet, GATA3, RORγt, HIF-1α). Primary outcomes focused on pathway-related gene expression differences between CA and non-CA groups. Secondary outcomes included comparisons of brain injury markers between CA and non-CA groups, as well as differences in JAK3-STAT5 pathway activity and biomarkers between ROSC and non-ROSC subgroups.

Results and Conclusions：No significant differences were observed in JAK3, STAT5a, or STAT5b expression between CA and non-CA groups, HIF-1α and GATA3 showed marked upregulation in CA patients. Reduced GATA3 and T-bet expression in ROSC subgroups, along with correlations between JAK3-STAT5 pathway genes and clinical parameters (diabetes comorbidity, initial PaO₂, serum albumin), suggest the pathway may modulate resuscitation outcomes through Th-cell differentiation and metabolic regulation. These findings offer novel insights into post-cardiac arrest immune-metabolic dysregulation and highlight potential targets for personalized immunotherapy.