Eriodictyol accelerates glucocorticoid-induced apoptosis by promoting osteoblast autophagy

doi:10.12307/2024.459

Abstract

Abstract: BACKGROUND: Glucocorticoid-induced osteoporosis is a common complication of systemic glucocorticoid therapy, which is mainly characterized by its inhibitory effect on osteoblasts. Eriodictyol inhibits osteoclast differentiation and osteoporosis-induced by ovariectomy. However, it is unclear whether eriodictyol regulates glucocorticoid-induced osteoblasts.
OBJECTIVE: To explore whether eriodictyol plays a role in glucocorticoid-induced osteoblast apoptosis and its potential regulatory mechanisms.
METHODS: Dexamethasone-pretreated osteoblasts MC3T3‐E1 were treated with the different concentrations (0, 0.5, 1, 2.5, 5, 10 μmol/L) of eriodictyol or 5 μmol/L 3-methyladenine, an autophagy inhibitor, and then transfected with heme oxygenase 1 overexpression vector (pcDNA-HMOX1) and empty vector (pcDNA vector). Cell proliferation and apoptosis were assessed by using cell counting kit-8 assay and flow cytometry, respectively. The activity of caspase-3 was detected with ELISA. Western blot assay was used to detect the protein expression of autophagy-related proteins LC3-II/LC3-I, p62, Atg5 and Atg12, the expression of apoptotic related proteins Bax and Bcl-2, as well as the protein expression of AMPK and p-AMPK.
RESULTS AND CONCLUSION: Low concentrations of eriodictyol were non-toxic to MC3T3-E1 cells and promoted cell proliferation, as well as increased the expression of autophagy related proteins LC3-II/LC3-I, p62, Atg5 and Atg12, decreased caspase-3 enzyme activity, inhibited Bax protein expression, promoted Bcl-2 protein expression and reduced dexamethasone-induced apoptosis in MC3T3-E1 cells in a dose-dependent manner. Moreover, eriodictyol significantly promoted heme oxygenase 1 expression in osteoblasts, whereas overexpression of heme oxygenase 1 promoted AMPK phosphorylation, activated autophagy, and inhibited dexamethasone-induced osteoblast apoptosis. While 3-methyladenine treatment counteracted the effects of heme oxygenase 1 overexpression on MC3T3-E1 cells. To conclude, low concentration of Eriodictyol is non-toxic to osteoblasts and activates AMPK signaling pathway by upregulating the expression of heme oxygenase 1, thereby promoting autophagy and inhibiting dexamethasone-induced osteoblast apoptosis. Eriodictyol has great potential for the treatment of glucocorticoid-induced osteoporosis.

Key words: eriodictyol, dexamethasone, heme oxygenase 1, MC3T3-E1 cell, the AMPK pathway

CLC Number:

Ma Yunfeng, Han Xiaofei. Eriodictyol accelerates glucocorticoid-induced apoptosis by promoting osteoblast autophagy[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(28): 4498-4504.

Figures/Tables 6

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