Assessment of progression of pulmonary fibrosis based on metabonomics and analysis of intestinal microbiota

Abstract

The main purpose of this study was to explore the changes of biomarkers in different developmental stages of bleomycin-induced pulmonary fibrosis (PF) in rats via comprehensive pathophysiology, UPLC-QTOF/MS metabonomic technology, and 16S rRNA gene sequencing of intestinal microbiota. The rats were randomly divided into normal control and 1-, 2- and 4-week model group. The rat model of PF was established by one-time intratracheal instillation of bleomycin. The levels of inflammatory and fibrosis-related factors such as hydroxyproline (HYP), type III procollagen (COL-III), type IV collagen (COL-IV), hyaluronidase (HA), laminin (LN), interleukin (IL)-1β, IL-6, malondialdehyde (MDA) increased and superoxide dismutase (SOD) decreased as the PF cycle progressed. In the 1-, 2- and 4-week model group, 2, 19 and 18 potential metabolic biomarkers and 3, 16 and 12 potential microbial biomarkers were detected, respectively, which were significantly correlated. Glycerophospholipid metabolism pathway was observed to be an important pathway affecting PF at 1, 2 and 4 weeks; arginine and proline metabolism pathways significantly affected PF at 2 weeks. Linoleic acid metabolism pathway exhibited clear metabolic abnormalities at 2 and 4 weeks of PF, and alpha-linolenic acid metabolism pathway significantly affected PF at 4 weeks.

KEY MESSAGES

In this study, metabolomics technology and intestinal microbiota 16S rRNA gene sequencing were used to search for biomarkers with significant differences in each stage of pulmonary fibrosis. Finally, the variation characteristics of each stage of the disease were discussed. The hope is to provide new insights into the development of diagnostic biomarkers and potential therapeutic targets at all stages.

GRAPHICAL ABSTRACT

Keywords:

• Pulmonary fibrosis
• disease progression
• metabonomics
• intestinal microbiota
• biomarker

Author contributions

Conceptualization, Jia-qi Liu, Hong-bing Zhou, Hong Chang and Song-li Shi; Data curation, Hong-bing Zhou, Jia Wang and Qian Li; Funding acquisition, Song-li Shi; Methodology, Hong-bing Zhou and Wan-fu Bai; Project administration, Song-li Shi; Writing – original draft, Jia-qi Liu, Hong Chang and Song-li Shi; Data graph making, Li-ya Fan and Jia-qi Liu; Writing – review & editing, Hong Chang, Song-li Shi, Li-ya Fan. All authors have read and agreed to the published version of the manuscript.

Disclosure statement

No potential conflict of interest was reported by the authors.

Institutional review board statement

The study was conducted in accordance with the Declaration of Helsinki and approved by Animal Ethics Committee of Baotou Medical College, Inner Mongolia University of Science and Technology (20170317).

Data availability statement

All data supporting the findings of this study are included in the article and its Supplementary Information files.

Additional information

Funding

This work was supported by the National Natural Science Foundation of China (81760782,81641137, 82060784), the “Grassland Talents” Youth Innovation and Entrepreneurship Talent Project of Inner Mongolia Autonomous Region, China (Q2017046), the 11th “Grassland Talents” Talent Project of Inner Mongolia Autonomous Region, China [(2021)8], the Natural Science Foundation of Inner Mongolia Autonomous Region, China (2019MS08189, 2023MS08022, 2023MS08029) the Scientific Research Project of Inner Mongolia Autonomous Region, China (NJZY21048), the Project of Baotou Medical College Innovation Team Development, China (bycxtd-02), the Project of Baotou Medical College Bud project, China (2021-56). The Inner Mongolia Autonomous Region postgraduate research innovation project (S20231236Z); the Program for Young Talents of Science and Technology in Universities of Inner Mongolia Autonomous Region (NTYT24079).