Tingting Li, Qianhui Sun, Luda Feng, Dong Yan, Boyuan Wang, Mingxuan Li, Xuejiao Xiong, Dayong Ma, Ying Gao
Abstract
Growing evidence has indicated that the characteristics of gut microbiota are associated with acute ischemic stroke (AIS). Phlegm-heat syndrome (PHS), a specific pathological state of the AIS, is one of the common traditional Chinese syndromes of stroke. The long duration of PHS in patients with AIS could lead to poor clinical outcomes. Gut microbiota characteristics in patients with both AIS and PHS, and their relationship remains unknown. This study was designed to investigate the alterations in gut microbiota in patients with AIS and PHS through a cross-sectional study. Fecal samples were collected from 10 patients with AIS and non-PHS (ntAIS), 7 patients with AIS and PHS (tAIS), and 10 healthy controls (HC). Samples were profiled via Illumina sequencing of the 16S rRNA V3-V4. Stroke severity was assessed at admission by the National Institutes of Health Stroke Scale (NIHSS) and modified Rankin scale (mRS); their correlation with gut microbiota was investigated.
Introduction
Stroke is the second-leading cause of death and disability worldwide; one out of four individuals will experience stroke at least once in their lifetime [1]. Stroke could be categorized into two types: ischemic and hemorrhagic stroke. It was estimated that approximately 62.4% of all stroke events were ischemic in 2009 [2]. The typical clinical manifestation of AIS is the neurological deficit over a single cerebral arterial vascular territory [3]. Additionally, at least half of the patients experienced gastrointestinal complications after stroke, including intestinal motility dysfunction, intestinal flora disturbance, leaky gut, intestinal bleeding, and enteropathogenic sepsis [4].
Materials and methods
Fecal samples were collected from patients with AIS and non-PHS (ntAIS group, n = 10); AIS and PHS (tAIS group, n = 7); and healthy controls (HC group, n = 10). Recruitment began in November 2021 and continued until the end of January 2022. Eligible subjects were male or female, from the age of 18–85 years. The baseline characteristics of the three groups were consistent and comparable, including age, sex, the body mass index, and comorbidities in order to exclude the influence of specific comorbidities (hypertension, diabetes, and hyperlipidemia) and other confounders that affect gut microbiota (Table 1; all P > 0.05). All patients with AIS within seven days of symptom onset were diagnosed according to the guidelines and confirmed by magnetic resonance imaging or computed tomography.
Results
We obtained 437 OTUs based on the 16S rRNA gene sequencing analysis (S2 Table). The tAIS, ntAIS, and HC groups had 416, 426, and 423 OTUs, respectively. Three and seven OTUs were specific to the ntAIS and HC groups, respectively.
Discussion
Microbiota are found in all multicellular organisms, including in the human gut, where they mutualistically support host health regulation. Changes in the gut microbiota may be a risk factor and contribute to AIS. Furthermore, stroke can lead to microbiome dysbiosis. Interactions between intestinal microbiota and AIS with a certain TCM syndrome, such as PHS, is an emerging focus for research. The shannon index were higher in tAIS group than those in ntAIS group with statistically significant difference. Additionally, the chao1 index were significantly higher in the tntAIS group than those in HC group.
Conclusion
This study characterized the alterations in gut microbiota composition in patients with tAIS and ntAIS. We found that the genera, Ruminococcaceae_ UCG_002 and Christensenellaceae_R-7_group could be used as indicators for discriminating PHS in AIS, as well as being potential therapeutic targets or underlying advantages of TCM treatment.
Citation: Li T, Sun Q, Feng L, Yan D, Wang B, Li M, et al. (2022) Uncovering the characteristics of the gut microbiota in patients with acute ischemic stroke and phlegm-heat syndrome. PLoS ONE 17(11): e0276598. https://doi.org/10.1371/journal.pone.0276598
Editor: Vasu D. Appanna, Laurentian University, CANADA
Received: April 10, 2022; Accepted: October 10, 2022; Published: November 3, 2022
Copyright: © 2022 Li et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Data Availability: The data are all contained within the Supporting Information files.
Funding: This research was financially supported by National Key R&D program “Evidence-based evaluation and mechanism of Chinese medicine intervention programs in the acute phase of stroke disease (Grant no.2018YFC1705000);the Dongzhimen hospital Beijing University of Chinese Medicine Project (Grant no. DZMKJCX-2020-003).The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Abbreviations: AIS, Acute Ischemic Stroke; PHS, Phlegm-Heat Syndrome; tAIS, Acute Ischemic Stroke and Phlegm-Heat Syndrome; ntAIS, Acute Ischemic Stroke and non-Phlegm-Heat Syndrome; HC, Healthy Controls; NIHSS, National Institutes of Health Stroke Scale; mRS, modified Rankin Scale; DSSEIS, Diagnostic Scale of Syndrome Elements in Ischemic Stroke; OTU, Operational Taxonomic Unit; PCoA, Principle Coordinates Analysis; LEfSE, Linear discriminant analysis Effect Size; FDR, false discovery rate; LDA, linear discriminant analysis.
