Age-Specific Pyroptosis Biomarker Analysis and Non-Pharmacological Intervention in Acute Respiratory Distress Syndrome

Yu Chen; Tong Liu; Yanpeng Zeng; Yifan Chen; Qiao Zou; Yurong Kong; Yangmin Li; Guantong Shen; Xin Wang; Yan Li

doi:10.3791/71443

Research Article

Age-Specific Pyroptosis Biomarker Analysis and Non-Pharmacological Intervention in Acute Respiratory Distress Syndrome

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DOI:

10.3791/71443

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June 5th, 2026

Yu Chen^*¹ , Tong Liu^*² , Yanpeng Zeng^*³ , Yifan Chen⁴ , Qiao Zou⁵ , Yurong Kong⁵ , Yangmin Li¹ , Guantong Shen³ , Xin Wang⁶ , Yan Li¹

¹Dongzhimen Hospital, Beijing University of Chinese Medicine, ²Third Affiliated Hospital, Beijing University of Chinese Medicine, ³Fangshan Hospital, Beijing University of Chinese Medicine, ⁴Xiamen Medical College, ⁵Beijing Huairou Hospital of Traditional Chinese Medicine, ⁶Guangdong Junfeng Spectrum Co., Ltd

^* These authors contributed equally

Summary

This protocol outlines an integrated bioinformatic and machine-learning workflow to identify age-stratified pyroptosis biomarkers in acute respiratory distress syndrome (ARDS), coupled with an in vivo methodology using an LPS-induced rat model to experimentally validate the preliminary efficacy of spectrum energy water and far-infrared radiation.

Abstract

Acute respiratory distress syndrome (ARDS) is a life-threatening inflammatory lung disorder with high morbidity and mortality, in which pyroptosis has emerged as a pivotal pathogenic mechanism. Considering that age-related immune and molecular differences may influence pyroptosis and therapeutic response, this study aimed to identify age-specific pyroptosis-associated biomarkers and evaluate the therapeutic potential of spectrum energy water (SEW) combined with far-infrared radiation (FIR). The protocol's comprehensive workflow encompasses transcriptomic dataset processing, age stratification, machine learning, and immune infiltration analysis to identify age-specific hub genes. This computational phase is followed by in vivo experimental validation using an LPS-induced ARDS rat model, employing histological assessment, ELISA, and Western blotting to rigorously evaluate the SEW+FIR intervention and verify hub gene expression. Sixteen pyroptosis-associated genes were identified, among which AXL and GSDME were predominantly associated with ARDS severity in older patients, whereas SPP1 was more relevant in younger individuals. Distinct immune signatures were observed, with M2 macrophage enrichment and immunosuppression in older patients, contrasted by pro-inflammatory activation in younger ones. Functional analyses implicated these genes in metabolic, inflammatory, and immune regulatory pathways. In vivo, SEW+FIR treatment alleviated lung injury, suppressed the production of inflammatory cytokines (IL-1β, IL-18, IL-6, TNF-α), and modulated the expression of AXL, GSDME, and SPP1. Collectively, these findings underscore age-dependent differences in pyroptosis-related mechanisms in ARDS and identify AXL, GSDME, and SPP1 as preliminary biomarker candidates that warrant further clinical validation. In the current murine experimental model, SEW+FIR demonstrated protective effects by alleviating systemic inflammation and modulating pyroptosis-related signaling, providing foundational in vivo support for its potential as a non-pharmacological adjunctive strategy.

Introduction

Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a severe respiratory disease characterized by an acute onset of severe hypoxemia and non-cardiogenic pulmonary edema, primarily resulting from diffuse alveolar damage¹. It can be triggered by a range of pulmonary and extrapulmonary insults. Pulmonary causes include infectious pneumonia, aspiration of gastric contents, and severe thoracic trauma, while extrapulmonary triggers arise from systemic inflammatory responses secondary to non-pulmonary insults, such as sepsis, pancreatitis, non-thoracic trauma, severe burns, massive transfusion, and reperfusion injury following lung t....