Integrative Multi-omics Analysis of Buti Huatan Tang in Chronic Obstructive Pulmonary Disease

Summary

Multi-omics profiling of Buti Huatan Tang (BTHTT) in chronic obstructive pulmonary disease reveals its potential to modulate the IL-1R2/IL-1β-thyroid hormone metabolic axis. This work provides a framework for further mechanistic validation of the bioactive components of BTHTT, establishing a foundation for future causal intervention studies required to confirm this axis.

Abstract

This study utilized a multi-omics and computational biology framework to investigate the therapeutic potential of the Traditional Chinese Medicine (TCM) formula Buti Huatan Tang (BTHTT) against chronic obstructive pulmonary disease (COPD). Significant physiological improvements were observed in a rat model following BTHTT intervention. Histological analysis showed a reversal of lung pathological damage, while biochemical assays, and transcriptomics confirmed the normalization of IL-1β and IL-1R2 levels. Additionally, metabolic profiling revealed that BTHTT corrected disruptions in T3 and T4 thyroid hormone levels. A negative correlation was observed between the IL-1β/IL-1R2 axis and these thyroid hormones, indicating that their regulation is associated with the formula’s therapeutic effect. Beyond direct measurements, machine learning algorithms identified ten COPD signature genes from clinical databases. Pathway enrichment analysis suggests that BTHTT may act through cytokine-cytokine-receptor interactions and thyroid hormone synthesis pathways. Furthermore, while 283 components were identified in vivo, compounds such as tanshinone IIA and cryptotanshinone are currently considered candidate active substances. Their role as primary drivers is supported by a model in which they stably bind to IL-1R2; this inference is based on molecular docking and molecular dynamics (MD) simulations rather than direct experimental isolation. Overall, the data support a model in which BTHTT exerts a multi-target effect on COPD by modulating inflammation and metabolic homeostasis. This integrated approach provides a refined scientific basis for the clinical application of BTHTT and highlights specific pathways for future experimental validation.

Introduction

Chronic obstructive pulmonary disease (COPD), one of the most common respiratory diseases in clinical practice, has become the third leading cause of death globally due to its extremely high disability rate and recurrent acute exacerbations¹. COPD was a significant factor leading to the loss of labor capacity in the population and a sharp increase in family care costs, with social and economic losses far exceeding those of most chronic diseases². In the “Healthy China Initiative 2030,” COPD has been listed as a key disease for prevention and treatment. Due to an incomplete understanding of the mechanisms unde....

Protocol

NOTE: The reagents and the equipment used in this study are listed in the Table of Materials.

Clinical data acquisition and preprocessing
To investigate the molecular signatures of COPD, transcriptomic datasets were retrieved from the Gene Expression Omnibus (GEO) database. A total of five datasets were selected: GSE11784, GSE12472, GSE16972, GSE38974, and GSE222965. The raw data and platform annotation files were downloaded for probe-to-gene mapping. When multiple probes targeted a single gene, the maximum expression value was retained. The resulting gene expression matrices (rows as genes, columns as samples) were m....

Results

Prediction of key characteristic genes in COPD
Following batch-effect correction of clinical data from multiple microarray platforms (Supplemental File 1–Supplemental Figure S1), a differential expression analysis was performed. 34 candidate genes (32 upregulated and 2 downregulated) were identified, including SRPX2, CLDN10, and TFF3 (Figure 1A).

To refine these candidates into a robust predi.......

Discussion

The synthesis and maintenance of thyroid hormone levels are significant for COPD patients and extend far beyond mere endocrine function, profoundly influencing disease progression, nutritional status, pulmonary function, and overall prognosis^31,32,33. Particularly during acute exacerbations, COPD patients often exhibited a hypothyroidism-like state referred to as “low T3 syndrome”. It was reported that decreased thyr.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Acetonitrile (chromatographic-grade)	Merck, Germany	1499230-935
Agilent 4150 bioanalyzer	Agilent Technologies, USA
Ammonium acetate	SIGMA, Germany	SRE0035-20230321
Angelica sinensis (Oliv.) Diels (Angelicae Sinensis Radix),	Guizhou Shanhaitongjia Pharmaceutical Co., Ltd., China	20240807
Astragalus membranaceus (Fisch.) Bge. (Astragali Radix),	Guizhou Shanhaitongjia Pharmaceutical Co., Ltd., China	20240601
ChemOffice		chemical structure drawing and molecular modeling software
Cinnamomum cassia (L.) J. Presl (Cinnamomi Cortex),	Guizhou Shanhaitongjia Pharmaceutical Co., Ltd., China	20240901
Eppendorf Centrifuge 5430 R	Eppendorf, Germany
Experimental cigarettes	Harbin Taihua, China	20230208
GraphPad Prism 8		statistical analysis and graphing software
Huangshan cigarettes		research-grade cigarettes for smoke exposure experiments
IL-1β ELISA kit	Shanghai Enzyme-Linked Biotechnology, China	E-EL-R0012c-20230522
Illumina Novaseq 6000 sequencing platform	Illumina, USA
Lepidium apetalum Willd. (Descurainiae Semen)	Guizhou Shanhaitongjia Pharmaceutical Co., Ltd., China	20240201
Lepidium sativum L. (Raphani Semen)	Guizhou Shanhaitongjia Pharmaceutical Co., Ltd., China	20240301
Lipopolysaccharide (LPS)	Shandong Huino Pharmaceutical, China	20230315
Mahonia fortunei (Lindl.) Fedde (Folium Mahoniae)	Guizhou Shanhaitongjia Pharmaceutical Co., Ltd., China	20240301
Methanol (chromatographic-grade)	Fisher, USA	DK745764
Nanodrop ND-2000 spectrophotometer	Thermo Scientific, USA
Perilla frutescens (L.) Britt. (Perillae Folium)	Guizhou Shanhaitongjia Pharmaceutical Co., Ltd., China	20240510
Pseudostellaria heterophylla (Miq.) Pax (Pseudostellariae Radix),	Guizhou Shanhaitongjia Pharmaceutical Co., Ltd., China	20250201
Qubit 4.0 fluorometer	Thermo Fisher Scientific, USA
RNA library construction kit	ABclonal, USA	RK20301-20230415
SMINA		molecular docking software
Salvia miltiorrhiza Bge. (Salviae Miltiorrhizae Radix et Rhizoma)	Guizhou Shanhaitongjia Pharmaceutical Co., Ltd., China	20240902
Small animal spontaneous activity recorder (KW-ZF)	Nanjing Karlvin Biotechnology, China
Sodium pentobarbital	Shanghai Experimental Reagent Procurement Company, China	20230412
TNF-α ELISA kit	Shanghai Enzyme-Linked Biotechnology, China	E-EL-R2856c-20230522
Traditional Chinese Medicine MS database (Shanghai Applied Protein Technology Co., Ltd.)		commercial traditional Chinese medicine mass spectrometry spectral database
Trizol reagent	Magen, China	20230508
UPLC BEH Amide column		amide-based UHPLC analytical column
UPLC HSS T3 column		reversed-phase UHPLC analytical column (T3-type)
Vanquish Neo UHPLC-Orbitrap Exploris 480 MS system	Thermo Scientific, USA
Vanquish UHPLC-Q-Exactive HF-X MS system	Thermo Scientific, USA