Journal of Shanghai Jiao Tong University (Medical Science) >

2021 , Vol. 41 >Issue 2: 147 - 153

DOI: https://doi.org/10.3969/j.issn.1674-8115.2021.02.004

Basic research

Effects of exosomes derived from human umbilical vein endothelial cells on apoptosis of pre-chondrogenic cells stimulated by inflammatory factors

  • Run-ze YANG ,
  • Wen-ning XU ,
  • Huo-liang ZHENG ,
  • Sheng-dan JIANG
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  • Department of Clinic of Spine Center, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China

Received date: 2020-05-28

  Online published: 2021-02-28

Supported by

National Natural Science Foundation of China(81672206);Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant Support(20181809)

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Abstract

Objective

·To investigate the effect of exosomes derived from umbilical vein endothelial cells (HUVECs) on apoptosis of murine pre-chondrogenic cell line ATDC5 cells under inflammatory stimulation.

Methods

·The exosomes derived from HUVECs were isolated by using an exosome isolation kit. Western blotting was used to detect the exosome marker proteins, including tumor susceptibility gene 101 (Tsg101), cluster differentiation 9 (CD9) and apoptosis linked gene-2-interacting protein X (Alix). The morphology of exosomes was observed by transmission electron microscope, and the size of exosomes was identified by particle size detection. Fluorescence microscope was used to observe the ATDC5 cell uptake of exosomes and the production of reactive oxygen species (ROS). TUNEL staining and flow cytometry were used to examine the effect of exosomes on ATDC5 cell apoptosis stimulated by interleukin-1β (IL-1β). Western blotting was used to detect the effect of exosomes on the expression levels of ATDC5 apoptosis-related proteins such as B-cell lymphoma/leukemia 2 (Bcl-2), Bcl-2 associated X protein (Bax), cleaved caspase-3 (c-caspase-3) and anti-oxidative stress-related proteins such as nuclear factor E2 related factor 2 (Nrf-2), Kelch-like ECH-associated protein 1 (Keap-1), heme oxygenase 1 (HO-1) and NADPH quinone oxidoreductase-like protein 1 (NQO-1) under IL-1β stimulation.

Results

·Under the transmission electron microscope, the HUVEC-derived exosomes were oval, hollow, double-layered, and positively expressed exosome markers CD9, Alix and Tsg101. Compared with the ATDC5 cells stimulated by IL-1β, ATDC5 cells stimulated by IL-1β incubated with exosomes had higher level of ROS (P=0.000) and higher apoptosis rate (P=0.000). The expression of Bax, c-caspase-3 and Keap-1 increased, and the expression of Bcl-2, Nrf-2, HO-1 and NQO-1 decreased in ATDC5 cells exposed to IL-1β and exosomes compared to ATDC5 cells only exposed to IL-1β.

Conclusion

·HUVEC-derived exosomes may promote ATDC5 cells apoptosis under the stimulation of IL-1β by inhibiting the ability of ATDC5 cell to resist oxidative stress.

Key words： exosome; osteoarthritis (OA); pre-chondrogenic cell; human umbilical vein endothelial cell (HUVEC); oxidative stress; apoptosis

Cite this article

Run-ze YANG , Wen-ning XU , Huo-liang ZHENG , Sheng-dan JIANG . Effects of exosomes derived from human umbilical vein endothelial cells on apoptosis of pre-chondrogenic cells stimulated by inflammatory factors[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2021 , 41(2) : 147 -153 . DOI: 10.3969/j.issn.1674-8115.2021.02.004

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