Mechanism by which exosomes from human fetal placental mesenchymal stem cells protect lung epithelial cells against oxidative stress injury

doi:10.3969/j.issn.2095-4344.3537

Abstract

Abstract:

BACKGROUND: Previous studies have confirmed that the culture supernatant of human fetal placental mesenchymal stem cells can protect lung epithelial cells from oxidative damage by inhibiting apoptosis.

OBJECTIVE: To investigate the protective effect of exosomes derived from human fetal placental mesenchymal stem cells on oxidative stress injured lung epithelial cells BEAS-2B.
METHODS: BEAS-2B cells were cultured in vitro with DME/F-12 complete medium containing different concentrations (100, 200, 300, 400, and 500 μmol/L) of H2O2 for 4 hours. Cells were cultured with DME/F-12 complete medium containing 300 μmol/L H2O2 for 2, 4, and 6 hours. Cell viability was measured by CCK-8 assay to determine the conditions of oxidative damaged model. Hoechst33258 staining was used to observe apoptosis. Q-PCR and western blot assay were used to detect the gene and protein expression of bax and bcl-2 to determine the validity of the model. The culture supernatant of human fetal placental mesenchymal stem cells from P3 was collected and the exosomes were isolated in accordance with exosome extraction kit instructions. The exosomes were treated on oxidative damaged BEAS-2B cells, and the expression levels of bax, bcl-2, nrf2 and keap1 were identified by q-PCR and western blot assay.
RESULTS AND CONCLUSION: (1) When BEAS-2B cells were treated with 300 μmol/L H2O2 for 4 hours, and the cell viability was (51.96±2.17)%, which could be used as the optimum condition for oxidative damaged model. Hoechst33258 staining, q-PCR and western blot assay showed that the model was effective. (2) Compared with the injury group, the expression of bax significantly decreased and the expression of bcl-2 significantly increased in exosomes group; the expression of nrf2 significantly increased and the expression of keap1 significantly decreased in exosomes, with significant differences (P < 0.05). (3) The results indicate that the exosomes derived from human fetal placental mesenchymal stem cells can inhibit the apoptosis of BEAS-2B cells induced by oxidative damage, and its mechanism may be related to the activation of Nrf2-Keap1-ARE signaling pathway.

Key words: stem cells, human fetal placental mesenchymal stem cells, lung epithelial cells, BEAS-2B, exosome, oxidative damage, cell apoptosis, signaling pathway

CLC Number:

Yan Xiurui, Tao Jin, Liang Xueyun. Mechanism by which exosomes from human fetal placental mesenchymal stem cells protect lung epithelial cells against oxidative stress injury[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(19): 2994-2999.

Figures/Tables 8

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