Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Diabetes. Mar 15, 2023; 14(3): 255-270
Published online Mar 15, 2023. doi: 10.4239/wjd.v14.i3.255
Characterization of gut microbial and metabolite alterations in faeces of Goto Kakizaki rats using metagenomic and untargeted metabolomic approach
Jin-Dong Zhao, Min Sun, Yan Li, Chan-Juan Yu, Ruo-Dong Cheng, Si-Hai Wang, Xue Du, Zhao-Hui Fang
Jin-Dong Zhao, Chan-Juan Yu, Ruo-Dong Cheng, Si-Hai Wang, Xue Du, Zhao-Hui Fang, Department of Endocrinology, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031, Anhui Province, China
Jin-Dong Zhao, Graduate School, Anhui University of Chinese Medicine, Hefei 230012, Anhui Province, China
Min Sun, School of Life Sciences, Anhui University, Hefei 230039, Anhui Province, China
Yan Li, Department of Infectious Disease, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031, Anhui Province, China
Author contributions: Zhao JD and Fang ZH participated in the design of the study and wrote the manuscript; Sun M, Li Y, Yu CJ, Cheng RD, Wang SH and Du X performed the experiment and helped complete the data analysis; The final version of the manuscript was reviewed and approved by all the authors.
Supported by the University Scientific Research Projects of Anhui, No. KJ2020A0401 and 2022AH050491; the open fund of the Ministry of Education Key Laboratory of Glucolipid Metabolic Disorder, No. GYDKFXM01; the Anhui University Collaborative Innovation Project, No. GXXT-2020-025; the National Natural Science Foundation of China, No. 82174153; the National Key Research and Development Program, No. 2018YFC1704202; the Anhui Provincial Quality Engineering Project of Universities, No. 2021jyxm0834; the Major and Difficult Diseases Project of Anhui Province, No. 2021zdynjb06; and the Clinical Research Project of Anhui University of Traditional Chinese Medicine, No. 2021yfylc01.
Institutional review board statement: All authors declare that Institutional Review Board approval was not applicable for this study because it did not involve human beings.
Institutional animal care and use committee statement: The experiments were approved by the Animal Ethics Committee of Anhui Chinese Medicine University (Hefei, China, No. ahucm-rats-2021133).
Conflict-of-interest statement: The authors declare that they have no conflicts of interest.
Data sharing statement: The data used to support the findings of this study are included within the article.
ARRIVE guidelines statement: The manuscript has been prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Zhao-Hui Fang, PhD, Chief Doctor, Department of Endocrinology, The First Affiliated Hospital of Anhui University of Chinese Medicine, No. 117 Meishan Road, Hefei 230031, Anhui Province, China. fangzhaohui1111@163.com
Received: December 2, 2022
Peer-review started: December 2, 2022
First decision: December 19, 2022
Revised: December 31, 2022
Accepted: February 7, 2023
Article in press: February 7, 2023
Published online: March 15, 2023
Processing time: 103 Days and 7.8 Hours
Abstract
BACKGROUND

In recent years, the incidence of type 2 diabetes (T2DM) has shown a rapid growth trend. Goto Kakizaki (GK) rats are a valuable model for the study of T2DM and share common glucose metabolism features with human T2DM patients. A series of studies have indicated that T2DM is associated with the gut microbiota composition and gut metabolites. We aimed to systematically characterize the faecal gut microbes and metabolites of GK rats and analyse the relationship between glucose and insulin resistance.

AIM

To evaluate the gut microbial and metabolite alterations in GK rat faeces based on metagenomics and untargeted metabolomics.

METHODS

Ten GK rats (model group) and Wistar rats (control group) were observed for 10 wk, and various glucose-related indexes, mainly including weight, fasting blood glucose (FBG) and insulin levels, homeostasis model assessment of insulin resistance (HOMA-IR) and homeostasis model assessment of β cell (HOMA-β) were assessed. The faecal gut microbiota was sequenced by metagenomics, and faecal metabolites were analysed by untargeted metabolomics. Multiple metabolic pathways were evaluated based on the differential metabolites identified, and the correlations between blood glucose and the gut microbiota and metabolites were analysed.

RESULTS

The model group displayed significant differences in weight, FBG and insulin levels, HOMA-IR and HOMA-β indexes (P < 0.05, P < 0.01) and a shift in the gut microbiota structure compared with the control group. The results demonstrated significantly decreased abundances of Prevotella sp. CAG:604 and Lactobacillus murinus (P < 0.05) and a significantly increased abundance of Allobaculum stercoricanis (P < 0.01) in the model group. A correlation analysis indicated that FBG and HOMA-IR were positively correlated with Allobaculum stercoricanis and negatively correlated with Lactobacillus murinus. An orthogonal partial least squares discriminant analysis suggested that the faecal metabolic profiles differed between the model and control groups. Fourteen potential metabolic biomarkers, including glycochenodeoxycholic acid, uric acid, 13(S)-hydroxyoctadecadienoic acid (HODE), N-acetylaspartate, β-sitostenone, sphinganine, 4-pyridoxic acid, and linoleic acid, were identified. Moreover, FBG and HOMA-IR were found to be positively correlated with glutathione, 13(S)-HODE, uric acid, 4-pyridoxic acid and allantoic acid and ne-gatively correlated with 3-α, 7-α, chenodeoxycholic acid glycine conjugate and 26-trihydroxy-5-β-cholestane (P < 0.05, P < 0.01). Allobaculum stercoricanis was positively correlated with linoleic acid and sphinganine (P < 0.01), and 2-methyl-3-hydroxy-5-formylpyridine-4-carboxylate was negatively associated with Prevotella sp. CAG:604 (P < 0.01). The metabolic pathways showing the largest differences were arginine biosynthesis; primary bile acid biosynthesis; purine metabolism; linoleic acid metabolism; alanine, aspartate and glutamate metabolism; and nitrogen metabolism.

CONCLUSION

Metagenomics and untargeted metabolomics indicated that disordered compositions of gut microbes and metabolites may be common defects in GK rats.

Keywords: Type 2 diabetes mellitus; Gut microbial; Metabolites; Goto-Kakizaki rats; Metagenomics; Untargeted metabolomics

Core Tip: Studies have suggested that the gut microbial and metabolites play an essential role in Goto Kakizaki rats. The results revealed evidence of a decrease in Prevotella sp. CAG:604 and increases in Lactobacillus_murinus and Allobaculum_stercoricanis. Fourteen potential metabolism biomarkers included glycochenodeoxycholic acid, uric acid, N-acetylaspartate, β-sitostenone, sphinganine, 4-pyridoxic acid, 13(S)-hydroxyoctadecadienoic acid, and linoleic acid, etc.