[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].

Li Zhang, Shiyuan Peng, Feiyang Tang, Jingwei Jian, Shuosheng Yuan, Xiaomei Xu
{"title":"[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].","authors":"Li Zhang, Shiyuan Peng, Feiyang Tang, Jingwei Jian, Shuosheng Yuan, Xiaomei Xu","doi":"10.7518/hxkq.2025.2024468","DOIUrl":null,"url":null,"abstract":"<p><strong>Objectives: </strong>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.</p><p><strong>Methods: </strong>The oxidative damage model of hPDLSCs was established using hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) andthe 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 H<sub>2</sub>O<sub>2</sub> 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.</p><p><strong>Results: </strong>H<sub>2</sub>O<sub>2</sub> 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 (<i>P<</i>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 (<i>P<</i>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 (<i>P<</i>0.05) and significantly diminished both the antioxidant effect of NAR and its ability to restore osteogenesis (<i>P<</i>0.05).</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":94028,"journal":{"name":"Hua xi kou qiang yi xue za zhi = Huaxi kouqiang yixue zazhi = West China journal of stomatology","volume":"43 4","pages":"559-569"},"PeriodicalIF":0.0000,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12419831/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Hua xi kou qiang yi xue za zhi = Huaxi kouqiang yixue zazhi = West China journal of stomatology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.7518/hxkq.2025.2024468","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

Objectives: 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) andthe 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).

Conclusions: 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.

[研究柚皮素通过调控叉头盒蛋白O-1/β-catenin通路对过氧化氢诱导的人牙周韧带干细胞氧化损伤的保护作用]。
目的:探讨柚皮素(naringenin, NAR)对氧化应激下人牙周韧带干细胞(hPDLSCs)成骨潜能的保护作用及其机制。方法:采用过氧化氢(H2O2)建立hPDLSCs氧化损伤模型,用不同浓度的NAR和0.5 μmol/L叉头盒蛋白O-1 (FOXO1)抑制剂AS1842856处理hPDLSCs。之后,使用细胞计数试剂盒-8 (CCK8)确定H2O2和NAR的最佳浓度。采用碱性磷酸酶(ALP)染色和实时荧光定量反转录聚合酶链反应(qRT-PCR)检测各组hPDLSCs中ALP、矮个子相关转录因子2 (RUNX2)和骨钙素(OCN)的表达。采用酶联免疫吸附法(ELISA)和2′,7′-二氯荧光素双乙酸酯(DCFH-DA)染色法检测hPDLSCs中活性氧(ROS)、丙二醛(MDA)和乳酸脱氢酶(LDH)的表达。同时,采用qRT-PCR和western blot检测FOXO1和β-catenin的表达水平,FOXO1和β-catenin均为通路相关基因和蛋白。结果:H2O2暴露导致hPDLSCs氧化损伤增加,表现为细胞内ROS水平升高,MDA和LDH表达增加(P0.05)。同时,hPDLSCs的成骨分化能力下降,表现为ALP染色变浅,成骨分化相关基因ALP、RUNX2、OCN表达水平降低(P0.05)。与NAR共处理可减轻hPDLSCs的氧化损伤,增强其抗氧化能力,恢复其成骨能力。FOXO1抑制剂AS1842856下调β-catenin的表达(P0.05),显著降低NAR的抗氧化作用和恢复成骨的能力(P0.05)。结论:NAR可通过激活hPDLSCs内FOXO1/β-catenin信号通路,增强hPDLSCs的抗氧化能力,从而减轻氧化应激损伤,减轻成骨能力的丧失。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信