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Diabetologia

pp 1–13 | Cite as

Mesenchymal stromal cell-derived exosomes ameliorate peripheral neuropathy in a mouse model of diabetes

  • Baoyan Fan
  • Chao Li
  • Alexandra Szalad
  • Lei Wang
  • Wanlong Pan
  • Ruilan Zhang
  • Michael Chopp
  • Zheng Gang Zhang
  • Xian Shuang LiuEmail author
Article

Abstract

Aims/hypothesis

Diabetic peripheral neuropathy (DPN) is one of the major complications of diabetes, which contributes greatly to morbidity and mortality. There is currently no effective treatment for this disease. Exosomes are cell-derived nanovesicles and play an important role in intercellular communications. The present study investigated whether mesenchymal stromal cell (MSC)-derived exosomes improve neurological outcomes of DPN.

Methods

Exosomes were isolated from the medium of cultured mouse MSCs by ultracentrifugation. Diabetic mice (BKS.Cg-m+/+Leprdb/J, db/db) at the age of 20 weeks were used as DPN models. Heterozygous mice (db/m) of the same age were used as the control. MSC-exosomes were administered weekly via the tail vein for 8 weeks. Neurological function was evaluated by testing motor and sensory nerve conduction velocities, and thermal and mechanical sensitivity. Morphometric analysis was performed by myelin sheath staining and immunohistochemistry. Macrophage markers and circulating cytokines were measured by western blot and ELISA. MicroRNA (miRNA) array and bioinformatics analyses were performed to examine the exosomal miRNA profile and miRNA putative target genes involved in DPN.

Results

Treatment of DPN with MSC-exosomes markedly decreased the threshold for thermal and mechanical stimuli and increased nerve conduction velocity in diabetic mice. Histopathological analysis showed that MSC-exosomes markedly augmented the density of FITC-dextran perfused blood vessels and increased the number of intraepidermal nerve fibres (IENFs), myelin thickness and axonal diameters of sciatic nerves. Western blot analysis revealed that MSC-exosome treatment decreased and increased M1 and M2 macrophage phenotype markers, respectively. Moreover, MSC-exosomes substantially suppressed proinflammatory cytokines. Bioinformatics analysis revealed that MSC-exosomes contained abundant miRNAs that target the Toll-like receptor (TLR)4/NF-κB signalling pathway.

Conclusions/interpretation

MSC-derived exosomes alleviate neurovascular dysfunction and improve functional recovery in mice with DPN by suppression of proinflammatory genes.

Keywords

Diabetes Diabetic peripheral neuropathy Exosomes Inflammation Mesenchymal stromal cells miRNA 

Abbreviations

BCA

Bicinchoninic acid

DPN

Diabetic peripheral neuropathy

GSK

Glycogen synthase kinase

IENF

Intraepidermal nerve fibre

iNOS

Inducible nitric oxide synthase

IPA

Ingenuity Pathway Analysis

IRAK1

Interleukin-1 receptor-associated kinase 1

MBP

Myelin basic protein

MCV

Motor nerve conduction velocity

miRNA

MicroRNA

MSC

Mesenchymal stromal cell

MSC-exosomes

MSC-derived exosomes

PGP9.5

Protein gene product 9.5

PU

Perfusion units

ROI

Region of interest

SCV

Sensory nerve conduction velocity

TLR

Toll-like receptor

Notes

Acknowledgements

The authors thank J. Landschoot-Ward and Q.-e. Lu (Department of Neurology, Henry Ford Health System) for immunohistochemistry staining.

Contribution statement

XSL and BF designed the study, analysed and interpreted data, and composed the manuscript. BF, CL, AS, LW, WP, RZ, MC and ZGZ conducted the experiments, acquired and analysed data and edited the manuscript. All authors have critically reviewed and approved the manuscript. XSL and ZGZ are the guarantors of this work and, as such, had full access to all of the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis.

Funding

This work was supported by the National Institutes of Health National Institute of Diabetes and Digestive and Kidney Disease grant RO1 RDK102861A (XSL), American Heart Association Grant-in-Aid 14GRNT20460167 (XSL), and RO1 NS075156 (ZGZ).

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2019_5043_MOESM1_ESM.pdf (512 kb)
ESM (PDF 511 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Baoyan Fan
    • 1
  • Chao Li
    • 1
  • Alexandra Szalad
    • 1
  • Lei Wang
    • 1
  • Wanlong Pan
    • 1
  • Ruilan Zhang
    • 1
  • Michael Chopp
    • 1
    • 2
  • Zheng Gang Zhang
    • 1
  • Xian Shuang Liu
    • 1
    Email author
  1. 1.Department of NeurologyHenry Ford Health SystemDetroitUSA
  2. 2.Department of PhysicsOakland UniversityRochesterUSA

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