Investigating the protective effect of naringenin on hydrogen peroxide induced oxidative damage of human periodontal ligament stem cells by regulating the forkhead box protein O-1/β-catenin pathway

doi:10.7518/hxkq.2025.2024468

Abstract

Abstract:

Objective Investigating the protective effect of naringenin (NAR) on the osteogenic potential of human periodontal ligament stem cells (hPDLSCs) under oxidative stress and its related mechanisms. Methods The oxidative damage model of hPDLSCs was established using hydrogen peroxide (H₂O₂) and_{the hPDLSCs were treated with different concentrations of NAR and 0.5 μmol/L forkhead box protein O-1 (FOXO1) inhibitor AS1842856. After that, the cell counting kit-8 (CCK8) was used to determine the optimal concentrations of H₂O₂ and NAR. The alkaline phosphatase (ALP) staining and real time fluorescent quantitative reverse transcription polymerase chain reaction (qRT-PCR) were employed to assess the expression of ALP, runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN) in hPDLSCs of each group. The enzyme-linked immunosorbent assay (ELISA) and 2’,7’-dichlorofluorescin diacetate (DCFH-DA) staining were utilized to evaluate the expression of reactive oxygen species (ROS), malondialdehyde (MDA) and lactate dehydrogenase (LDH) in hPDLSCs. Meanwhile, qRT-PCR and western blot were used to detect the expression levels of FOXO1 and β-catenin, both are pathway related genes and proteins. Results H₂O₂ exposure led to an increase in oxidative damage in hPDLSCs, characterized by a rise in intracellular ROS levels and increased expression of MDA and LDH (P<0.05). At the same time, the osteogenic differentiation ability of hPDLSCs decreased, as evidenced by lighter ALP staining and reduced expression levels of osteogenic differentiation-related genes ALP, RUNX2 and OCN (P<0.05). Co-treatment with NAR alleviated the oxidative damage in hPDLSCs, enhanced their antioxidant capacity, and restored their osteogenic ability. The FOXO1 inhibitor AS1842856 downregulated the expression of β-catenin (P<0.05) and significantly diminished both the antioxidant effect of NAR and its ability to restore osteogenesis (P<0.05). Conclusion NAR can enhance the antioxidant capacity of hPDLSCs by activating the FOXO1/β-catenin signaling pathway within hPDLSCs, thereby mitigating oxidative stress damage and alleviating the loss of osteogenic capacity.}

Key words: periodontal ligament stem cells, forkhead box protein O-1/β-catenin signaling pathway, oxidative stress, naringenin, osteogenic differentiation

CLC Number:

R781.4

Zhang Li, Peng Shiyuan, Tang Feiyang, Jian Jingwei, Yuan Shuosheng, Xu Xiaomei. Investigating the protective effect of naringenin on hydrogen peroxide induced oxidative damage of human periodontal ligament stem cells by regulating the forkhead box protein O-1/β-catenin pathway[J]. West China Journal of Stomatology, 2025, 43(4): 559-569.

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引物名称	序列
RUNX2	F：GAACCCAGAAGGCACAGACA
	R：GGCTCAGGTAGGAGGGGTAA
ALP	F：GCCGCCCGCTTTAACC
	R：GTCCCACTGACTTCCCTGCTT
OCN	F：TCCTTTGGGGTTTGGCCTAC
	R：CCAGCCTCCAGCACTGTTTA
FOXO1	F：ATTCGGTCATGCCAGCGTAT
	R：CAAGCGGTTCATGGCAGATG
β-catenin	F：CCTAGCTGGTGGACTGCAGA
	R：CACCACTGGCCAGAATGATGA
β-actin	F：CGACCCGTCTACAGGTGAAC
	R：GAGAGGGACTCAATCAGCCG