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The potential mechanism of Liu–Wei–Di–Huang Pills in treatment of type 2 diabetic mellitus: from gut microbiota to short-chain fatty acids metabolism

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Abstract

Background

Type 2 diabetes mellitus (T2DM) has already become a global pandemic. Recently, reports showed its pathogenesis was closely related to a disorder of gut microbiota. In China, the Liu–Wei–Di–Huang Pills (LWDH) have treated T2DM for thousands of years. However, its therapeutic mechanism associated with gut microbiota is worthy of further study.

Aims

This study aims to investigate the effects of LWDH on T2DM by regulating gut microbiota and short-chain fatty acids (SCFAs) in Goto–Kakizaki (GK) rats.

Methods

T2DM models were successfully established based on GK rats and administrated with LWDH. The changes in fasting blood glucose (FBG), oral glucose tolerance test (OGTT), and serum insulin (INS) were determined, and the immunohistochemical (IHC) method was used to test INS expression in pancreas. The 16S-ribosomal DNA (16S rDNA) sequencing analysis assessed gut microbiota structural changes; a gas chromatography–mass spectrometer (GC–MS)-based metabolomics method was adopted to detect SCFA levels. The pathological morphology of jejunum was detected by hematoxylin–eosin (H&E) staining, and the expression of GPR43, GPR41, GLP-1, and GLP-1R was evaluated by qRT-PCR and ELISA, respectively.

Results

We observed that GK rats treated with LWDH: (a) has altered the microbial structure and promoted the abundance of bacteria in Firmicutes, including Lactobacillus, Allobaculum, and Ruminococcus_2, (b) increased SCFAs levels involving acetic acid, propionic acid, and butyric acid and (c) alleviated T2DM and jejunum injuries potentially based on SCFAs-GPR43/41-GLP-1 pathway.

Conclusion

LWDH could improve T2DM by regulating gut microbiota and SCFAs, and the therapeutic mechanism might be related to the SCFAs-GPR43/41-GLP-1 pathway.

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Data availability

The data used to support the findings of this study are available from the corresponding author upon request.

Abbreviations

T2DM:

Type 2 diabetes mellitus

LWDH:

Liu–Wei–Di–Huang Pills

SCFAs:

Short-chain fatty acids

GK:

Goto–Kakizaki

FBG:

Fasting blood glucose

OGTT:

Oral glucose tolerance test

INS:

Serum insulin

IHC:

Immunohistochemical

16S rDNA:

The16S-ribosomal DNA

GC–MS:

A gas chromatography–mass spectrometer

H&E:

Hematoxylin-eosin

GLP-1:

Glucagon-like peptide-1

GLP-1R:

Glucagon-like peptide-1 receptor

GPCRs:

G-protein-coupled receptors

GPR43:

G-protein-coupled receptors 43

GPR41:

G-protein-coupled receptors 41

TCM:

Traditional Chinese medicine

MOD:

Model group

POS:

Positive group

CON:

Control group

AOD:

Average optical density

PCoA:

Principal coordinates analysis

NMDS:

Non-metric multidimensional scaling

LDA:

Linear discriminant analysis

LEfSe:

Linear discriminant analysis effect size

DC:

Degree

BC:

Betweenness centrality

CC:

Closeness centrality

PICRUSt 2.0:

Phylogenetic Investigation of Communities by Reconstruction of Unobserved States Ver2.0

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Acknowledgements

This work was supported by the National Key R&D Program of China (No:2018YFC1704400); National Nature Science Foundation of China (No. 82074400, U21A20411); Natural Science Foundation of Hunan Province (2020JJ5325); Key Research and Development Project of Hunan Province Science and Technology (2020ZYQ038, 2020SK2029, 2020SK2101); Project of Changsha Technology Innovation Center (kh2004018); Training Program for Excellent Young Innovators of Changsha (kq1802017); Research Project of Traditional Chinese Medicine Bureau of Hunan Province (202092); Program of Survey of Chinese Medicines of China ([2017]66), Research on the Comprehensive Development and Utilization of Characteristic Traditional Chinese Medicine Resources (2060302). China Post-doctoral Science Foundation (2022M711131)

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Authors and Affiliations

  1. Hunan Academy of Chinese Medicine, Hunan University of Chinese Medicine, Changsha, 410013, Hunan, People’s Republic of China

    Zi-yang Yi, Lin Chen, Dan He, Di Zhao, Shui-han Zhang, Rong Yu & Jian-hua Huang

  2. Hunan Key Laboratory of TCM Prescription and Syndromes Translational Medicine, Hunan University of Chinese Medicine, Changsha, 410208, Hunan, People’s Republic of China

    Rong Yu & Jian-hua Huang

  3. H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan

    Yan Wang

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  1. Zi-yang Yi
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  2. Lin Chen
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  3. Yan Wang
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  4. Dan He
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  5. Di Zhao
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Contributions

DH, YW designed the experiments, supervised, and participated in the entire work. ZYY, LC were in charge of statistical analysis and wrote the manuscript and revised the manuscript. DZ maintained and performed animal studies. SHZ participated in study design and data collection. All the authors revised and approved the final manuscript.

Corresponding authors

Correspondence to Rong Yu or Jian-hua Huang.

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The authors declare that there is no conflict of interest regarding the publication of this paper.

Ethics statement

The animal study was reviewed and approved by The Animal Ethical Committee of Hunan Academy of Chinese Medicine.

Research Involving Human and Animal Rights

Our study did not involve human studies.

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Yi, Zy., Chen, L., Wang, Y. et al. The potential mechanism of Liu–Wei–Di–Huang Pills in treatment of type 2 diabetic mellitus: from gut microbiota to short-chain fatty acids metabolism. Acta Diabetol 59, 1295–1308 (2022). https://doi.org/10.1007/s00592-022-01922-y

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