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Human milk-derived extracellular vesicles alleviate high fat diet-induced non-alcoholic fatty liver disease in mice

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Abstract

Background

Nonalcoholic fatty liver disease (NAFLD), characterized by excessive hepatic lipid accumulation, imposes serious challenges on public health worldwide. Breastfeeding has been reported to reduce the risk of NAFLD. Extracellular vesicles (EVs) are bilayer membrane vesicles released from various cells into the extracellular space, participating in multiple life processes. Whether EVs from human milk exert metabolic benefits against NAFLD is worth investigating.

Methods and results

In this study, the EVs were isolated from human milk collected from healthy mothers and quantified. Functional analyses were performed using the NAFLD mouse model and free fatty acid (FFA)-stimulated mouse primary hepatocytes. The results showed that human milk-derived EVs could effectively alleviate high fat diet-induced hepatic steatosis and insulin resistance in mice with NAFLD via inhibiting lipogenesis and increasing lipolysis. The FFA-induced lipid accumulation was also inhibited in hepatocytes after treatment with human milk-derived EVs. Mechanistically, the human milk derived-EVs cargo (proteins and miRNAs), which linked to lipid metabolism, may be responsible for these beneficial effects.

Conclusion

The findings of this study highlighted the therapeutic benefits of human milk-derived EVs and provided a new strategy for NAFLD treatment.

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Abbreviations

ACC:

Acetyl-CoA carboxylase

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

ATGL:

Adipose triglyceride lipase

AUC:

Area under curve

BCA:

Bicinchoninic acid

BSA:

Bovine serum albumin

CCK-8:

Cell counting kit-8

CLU:

Clusterin

CPT1a:

Carnitine palmitoyl-transferase 1a

EVs:

Extracellular vesicles

FAS:

Fatty acid synthase

GO:

Gene ontology

GTT:

Glucose tolerance tests

FFA:

Free fatty acid

H&E:

Hematoxylin–eosin staining

HCC:

Hepatocellular carcinoma

HFD:

High-fat diet

HOMA-IR:

Homoeostasis model assessment of insulin resistance

HRP:

Horseradish peroxidase

i.p.:

Intraperitoneally

ITTs:

Insulin tolerance tests

KEGG:

Kyoto encyclopedia of genes and genomes

LTF:

Lactotransferrin

MFGE8:

Milk fat globule-epidermal growth factor-factor 8

NAFLD:

Nonalcoholic fatty liver disease

NASH:

Non-alcoholic steatohepatitis

NCD:

Normal chow diet

NEC:

Necrotizing enterocolitis

NTA:

Nanoparticle tracking analysis

SREBP-1C:

Sterol-regulatory element binding protein-1C

TEM:

Transmission electron microscopy

TC:

Total cholesterol

TG:

Triglyceride

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Acknowledgements

We thank all the lactating mothers who donated the milk for this study.

Funding

This work was funded by grants from the National Natural Science Foundation of China (Grant Nos. 81770866, 81770837, 81900783 and 82070879), Maternal and Infant Nutrition and Health Research Project (Grant No. 2021FY005), Jiangsu Provincial Medical Innovation Team (Grant no. CXTDA2017001), Jiangsu Provincial Key Research and Development Program (Grant Nos. BE2018616 and BK20211017), Jiangsu Natural Science Foundation (Grant No. BK20190139), Six Talent Peaks Project in Jiangsu Province (Grant No. YY-084), Specialized Disease Cohort Study of Nanjing Medical University (Grant No. NMUC2018014A), 333 High Level Talents Training Project of Jiangsu Province, and Science and Technology Development Foundation Item of Nanjing Medical University (Grant No. NMUB2020141).

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Authors and Affiliations

  1. Department of Children Health Care, Women’s Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing, Jiangsu, China

    Xue Jiang, Hong Zhong, Xiaoxiao Zhang, Li Liu, Xianwei Cui, Xia Chi & Chenbo Ji

  2. Nanjing Maternal and Child Health Institute, Women’s Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing, Jiangsu, China

    Xue Jiang, Yangyang Wu, Hong Zhong, Xiaoxiao Zhang, Li Liu, Xianwei Cui, Xia Chi & Chenbo Ji

  3. School of Pediatrics, Nanjing Medical University, Nanjing, Jiangsu, China

    Xue Jiang, Hong Zhong, Xiaoxiao Zhang, Xingzhen Sun, Li Liu, Xianwei Cui, Xia Chi & Chenbo Ji

Authors
  1. Xue Jiang
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  2. Yangyang Wu
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  3. Hong Zhong
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  8. Xia Chi
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Contributions

CJ and XC conceived and designed the project. XJ did the majority of the experiments and wrote the first draft of the paper. YW and HZ aided in experiments and editing. XZ performed data analysis of miRNA sequencing and mass spectrometry. XC collected the human milk and provided EVs. XS and LL provided expertise on experimental design and interpretation of data. XC, XC and CJ provided the funding and approved the final version of the manuscript.

Corresponding authors

Correspondence to Xianwei Cui, Xia Chi or Chenbo Ji.

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Conflict of interest

All the authors declare that no conflict of interest.

Ethical approval

First, all the donors had signed the informed consent for breast milk donation. The Institutional Review Board at Women’s Hospital of Nanjing Medical University approved this study (Permission Number 2020KY-007). Second, all animal experiments were approved by the Animal Ethics Committee of Nanjing Medical University (Permission Number IACUC-2011052). All experimental protocols followed the guidelines issued by National Institute of Health and the Institutional Animal Care and Use Committee of Nanjing Medical University.

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Jiang, X., Wu, Y., Zhong, H. et al. Human milk-derived extracellular vesicles alleviate high fat diet-induced non-alcoholic fatty liver disease in mice. Mol Biol Rep 50, 2257–2268 (2023). https://doi.org/10.1007/s11033-022-08206-2

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  • DOI: https://doi.org/10.1007/s11033-022-08206-2

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