Influence and mechanism of extracellular vesicles derived from human adipose-derived mesenchymal stem cells on pyroptosis of human umbilical vein endothelial cells induced by high glucose

Feng Junyun; Fei Xiao; Fang Shaoyihan; An Jingwen; Shi Yan; Liu Dewu

doi:10.3760/cma.j.cn501225-20240120-00025

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2024 > Accepted Manuscript

Feng Junyun,Fei Xiao,Fang Shaoyihan,et al.Influence and Mechanism of Extracellular Vesicles Derived from Human Adipose-Derived Mesenchymal Stem Cells on pyroptosis of Human Umbilical Vein Endothelial Cells induced by high glucose[J].Chin J Burns Wounds,2025,41(1):1-10.DOI: 10.3760/cma.j.cn501225-20240120-00025.

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Influence and mechanism of extracellular vesicles derived from human adipose-derived mesenchymal stem cells on pyroptosis of human umbilical vein endothelial cells induced by high glucose

doi: 10.3760/cma.j.cn501225-20240120-00025

1.
Medical Center of Burn Plastic and Wound Repair, the First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang330006, China
2.
Medical College of Nanchang University, Nanchang330006, China

Funds:

Regional Science Foundation Project of National Natural Science Foundation of China 82460447, 81460293

Chongqing Traditional Chinese Medicine Inheritance and Innovation Team Project 2023090006KJZX2022WJW008

More Information

Corresponding author: Liu Dewu, Email: dewuliu@126.com
Received Date: 2024-01-20
Available Online: 2024-12-27

Abstract

Abstract

Objective To investigate the influence of extracellular vesicles (EVs) derived from human adipose-derived mesenchymal stem cells (hADMSCs), i.e. hADMSC-EVs on pyroptosis of human umbilical vein endothelial cells (HUVECs) induced by high glucose and their mechanisms, with the aim of providing evidence for improving vascular dysfunction in diabetic wounds. Methods This study was an experimental research. Umbilical cords were collected from five lying-in women aged 25 to 40 years who underwent normal vaginal delivery at the Department of Obstetrics and Gynecology of the First Affiliated Hospital of Nanchang University. HUVECs were isolated and successfully identified. Adipose tissue was obtained from six healthy women aged 25 to 35 years who underwent abdomen liposuction at the Department of Plastic Surgery of the First Affiliated Hospital of Nanchang University. hADMSCs were isolated and successfully identified. hADMSC-EVs were extracted using high-speed centrifugation and successfully identified. The fourth passage of HUVECs were cultured in endothelial cell medium containing glucose in a morlarity of 33 mmol/L and divided into three groups: phosphate buffer solution (PBS) group cultured with PBS, EV group cultured with hADMSC-EVs, and EV+LY294002 group cultured with hADMSC-EVs and the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway inhibitor LY294002. Western blotting was used to detect the expression of PI3K/Akt signaling pathway-related proteins PI3K and Akt, and pyroptosis-related proteins nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase 1 (caspase-1), gasdermin D, interleukin-1β (IL-1β), and IL-18 after 48 hours of cell culture in each group. A cell counting kit-8 was used to test the proliferation levels of cells in each group at 0, 12, 24, 36, 48, 60, and 72 hours of culture. After 48 hours of cell culture in each group, the cell scratch test was performed and the cell migration rates at 12 and 24 hours after scratching were calculated; the cell Transwell assay was conducted and the number of cells migrating in 24 hours was calculated; the cell tube formation experiment was performed, and the total length of tube formation and the number of branch nodes were measured and counted. The sample size was three. Results After 48 hours of culture, the protein expressions of PI3K and Akt of cells in EV group were significantly higher than those in PBS group (P<0.05), and the protein expressions of PI3K and Akt of cells in EV+LY294002 group were significantly lower than those in EV group (P<0.05). After 48 hours of culture, the protein expressions of NLRP3, caspase-1, gasdermin D, IL-1β, and IL-18 of cells in EV group were 0.54±0.08, 0.96±0.11, 0.525±0.061, 1.216±0.039, and 1.317±0.023, respectively, which were significantly lower than 2.32±0.11, 1.86±0.07, 1.256±0.113, 2.589±0.084, and 2.042±0.132 in PBS group (P<0.05); the protein expressions of NLRP3, caspase-1, gasdermin D, IL-1β, and IL-18 of cells in EV+LY294002 group were 1.16±0.05, 1.37±0.06, 0.962±0.028, 1.834±0.017, and 1.803±0.065, respectively, which were significantly higher than those in EV group (P<0.05). At 12, 24, 36, 48, 60, and 72 hours of culture, the proliferation levels of cells in EV group were significantly higher than those in PBS group (P<0.05), and the proliferation levels of cells in EV+LY294002 group were significantly lower than those in EV group (P<0.05). After 48 hours of culture, the cell migration rates at 12 and 24 hours after scratching in EV group were significantly higher than those in PBS group (P<0.05), and the cell migration rates at 12 and 24 hours after scratching in EV+LY294002 group were significantly lower than those in EV group (P<0.05); the number of cells migrating in 24 hours in EV group was significantly greater than that in PBS group (P<0.05), and the number of cells migrating in 24 hours in EV+LY294002 group was significantly less than that in EV group (P<0.05). After 48 hours of culture, compared with those in PBS group, the total length of tube formation of cells in EV group was significantly longer (P<0.05), and the number of branch nodes was significantly increased (P<0.05); compared with those in EV group, the total length of tube formation in EV+LY294002 group was significantly shorter (P<0.05), and the number of branch nodes was significantly decreased (P<0.05). Conclusions hADMSC-EVs can inhibit the expression of pyroptosis-related proteins in HUVECs induced by high glucose through the PI3K/Akt signaling pathway and improve their proliferation, migration, and angiogenesis capabilities.
- Diabetes mellitus,
- Extracellular vesicles,
- Human umbilical vein endothelial cells,
- Pyroptosis,
- Phosphatidylinositol 3-kinase,
- Adipose-derived mesenchymal stem cells,
- Protein kinase B

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References

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