Glutamine regulates the effect of hormones on the apoptosis of bone microvascular endothelial cells

doi:10.12307/2026.660

Abstract

Abstract: BACKGROUND: Glucocorticoids can significantly inhibit the expression of proteins related to the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway in bone microvascular endothelial cells, thereby triggering programmed cell death and necrosis. Glutamine has been shown to activate the PI3K/Akt/mTOR signaling pathway.
OBJECTIVE: To explore how glutamine regulates the effect of glucocorticoids on the apoptosis of bone microvascular endothelial cells.
METHODS: Bone microvascular endothelial cells were extracted from the femoral head tissues of Sprague-Dawley rats under aseptic conditions. Passage 3 bone microvascular endothelial cells were cultured for 12 hours and then divided into four groups: the control group was routinely cultured for 24 hours, the glucocorticoid group was cultured with methylprednisolone sodium succinate for 24 hours, the glucocorticoid+glutamine group was cultured with methylprednisolone sodium succinate for 12 hours and then with glutamine for 12 hours, and the glucocorticoid+glutamine+LY294002 group was cultured with methylprednisolone sodium succinate for 12 hours followed by addition of glutamine treatment for 6 hours and the PI3K/Akt signaling pathway inhibitor LY294002 for 6 hours. After culture, cell apoptosis was detected by AO-PI double fluorescence staining and Annexin V/PI double staining by flow cytometry; BCL-2, Bax, PI3K, Akt, mTOR mRNA expression was detected by RT-qPCR; and western blot was used to detect BCL-2, Bax, PI3K, Akt, mTOR, p-PI3K, p-Akt, and p-mTOR protein expression.
RESULTS AND CONCLUSION: (1) The apoptosis rate was higher in the glucocorticoid group than in the control group and the glucocorticoid+glutamine group (P < 0.05), and the apoptosis rate was higher in the glucocorticoid+glutamine+LY294002 group than in the glucocorticoid+glutamine group (P < 0.05). (2) BCL-2, PI3K, Akt, mTOR mRNA expression in the glucocorticoid group was lower than that in the control group and glucocorticoid+glutamine group (P < 0.05), and Bax mRNA expression was higher than that in the control group and glucocorticoid+glutamine group (P < 0.05); BCL-2, PI3K, and mTOR expression in the glucocorticoid+glutamine+LY294002 group was lower than that in the glucocorticoid+glutamine group (P < 0.05), and Bax mRNA expression was higher than that in the glucocorticoid+glutamine group (P < 0.05). (3) BCL-2, PI3K, Akt, mTOR, p-PI3K, p-Akt, p-mTOR protein expression in the glucocorticoid group was lower than that in the control group and the glucocorticoid+glutamine group (P < 0.05), and Bax proteins were higher than that in the control group and the glucocorticoid+glutamine group (P < 0.05); BCL-2, PI3K, Akt, mTOR, p-PI3K, p-Akt, p-mTOR protein expression in the glucocorticoid+glutamine+LY294002 group was lower than that in the glucocorticoid+glutamine group (P < 0.05) and Bax protein expression was higher than that in the glucocorticoid+glutamine group (P < 0.05). To conclude, glutamine reduces glucocorticoid-induced apoptosis in bone microvascular endothelial cells through activation of the PI3K/Akt/mTOR signaling pathway.

Key words: glucocorticoid, apoptosis, steroid-induced osteonecrosis of the femoral head, glutamine, endothelial cells, LY294002, signaling pathway, femoral head

CLC Number:

Zan Yongfeng, Song Keguan, Liu Yuda. Glutamine regulates the effect of hormones on the apoptosis of bone microvascular endothelial cells[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(12): 2965-2974.

Figures/Tables 10

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