Programmed death receptor 1 inhibits osteogenic differentiation of rat bone marrow mesenchymal stem cells in a high glucose environment

doi:10.12307/2025.075

Abstract

Abstract: BACKGROUND: The mechanism of programmed death receptor-1 (PD-1) effect on osteogenic differentiation of bone marrow mesenchymal stem cells in high glucose environment remains unclear.
OBJECTIVE: To explore the effect of PD-1 on osteogenic differentiation of rat bone marrow mesenchymal stem cells in high glucose environment and its regulatory mechanism.
METHODS: Rat bone marrow mesenchymal stem cells were randomly divided into normal glucose group (5.6 mmol/L), high glucose group (30 mmol/L), PD-1 overexpression group, PD-1 overexpression no-load group, PD-1 knockdown group, PD-1 knockdown no-load group, and PI3K/AKT pathway inhibitor group (PD-1 knockdown+5 μmol/L LY294002). Rat bone marrow mesenchymal stem cells were cultured in high glucose to simulate the diabetic environment in vitro. The mRNA expression of PD-1 and ligand PD-L1 and the mRNA expression of osteogenic markers Runx2 and OSX in rat bone marrow mesenchymal stem cells were detected by qRT-PCR. The osteogenic differentiation ability was observed by alkaline phosphatase staining and alizarin red staining. Cell proliferation was detected by CCK-8 assay. The protein expressions of PD-1, PD-L1, p-PI3K, and p-AKT were detected by western blot assay.
RESULTS AND CONCLUSION: (1) The levels of PD-1 and PD-L1 were significantly increased in the high glucose environment in vitro, and the osteogenic differentiation ability of bone marrow mesenchymal stem cells was inhibited in the high glucose environment. (2) Knockdown of PD-1 expression could promote osteogenic differentiation of bone marrow mesenchymal stem cells, increase cell proliferation activity, and activate the PI3K/AKT pathway. (3) After addition of PI3K/AKT pathway inhibitor LY294002, the ability of bone marrow mesenchymal stem cells to differentiate into osteoblasts decreased. The results show that PD-1 is dependent on the PI3K/AKT signaling pathway to inhibit osteogenic differentiation of rat bone marrow mesenchymal stem cells under high glucose environment.

Key words: one marrow mesenchymal stem cell, high glucose, osteogenic differentiation, programmed death receptor-1, PI3K, AKT

CLC Number:

Han Nianrong, Huang Yifei, Akram · Osman, Liu Yanlu, Hu Wei . Programmed death receptor 1 inhibits osteogenic differentiation of rat bone marrow mesenchymal stem cells in a high glucose environment[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(19): 3961-3967.

Figures/Tables 2

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