口腔疾病防治2025，Vol.33Issue(9)：732-743,12.DOI:10.12016/i.issn.2096-1456.202550208
HIF-1α促进机械压力下牙周膜细胞的炎症应答反应
HIF-1α promotes the inflammatory response of periodontal ligament cells under mechanical stress
摘要
Abstract
Objective To investigate the molecular regulatory mechanism of hypoxia-inducible factor-1α(HIF-1α)in mechanical stress-induced inflammatory cytokine expression in human periodontal ligament cells(hPDLCs),provid-ing a theoretical basis and potential therapeutic target for inflammatory control during orthodontic treatment.Methods This study was approved by the Institutional Ethics Committee.Primary human periodontal ligament cells(hPDLCs)were isolated and cultured in vitro.Self-renewal capacity was confirmed via colony-forming assays,while osteogenic and adipogenic differentiation potential was evaluated via Alizarin Red S staining,alkaline phosphatase(ALP)activity as-says,and Oil Red O staining.An in vitro compressive force stimulation model(1.5 g/cm2,12 h)was established to com-pare inflammatory cytokine expression of hPDLCs—interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and HIF-1α—between the Control group(no mechanical stimulation)and the Force group(1.5 g/cm2,12 h)using quantitative real-time PCR(qRT-PCR),Western blot,and immunofluorescence(IF)staining.Mechanically in-duced HIF-1α-regulated gene expression changes were analyzed through transcriptomic sequencing.To explore pharma-cological inhibition,the small-molecule HIF-1α inhibitor LW-6 was applied at varying concentrations(10 μ mol/L,30 μmol/L,50 μmol/L)to optimize the treatment dose.Subsequently,qRT-PCR,Western blot,and IF staining were conducted to evaluate inflammatory cytokine of hPDLCs and HIF-1α expression in three groups:Control(no force),Force(1.5 g/cm2,12 h),and Force+LW6(1.5 g/cm2,12 h+30 μmol/L LW-6).Results Primary hPDLCs demon-strated self-renewal capacity along with osteogenic and adipogenic differentiation potential.Compared to the Control group,the Force group exhibited significantly increased mRNA and protein expression levels of inflammatory cytokines IL-1β,IL-6,and TNF-α,along with enhanced fluorescence intensity of IL-1β and TNF-α.Transcriptomic analysis re-vealed that mechanical compressive force activated the HIF-1 signaling pathway,which subsequently mediated inflam-matory responses and bone remodeling processes in hPDLCs.Furthermore,the mRNA and protein levels of HIF-1α were considerably elevated in the Force group compared to the Control group.Treatment with LW-6(10,30,or 50 μmol/L)effectively suppressed HIF-1α expression,with 30 μmol/L LW-6 identified as the optimal concentration for intervention.In subsequent experiments,the Force group showed significant upregulation in mRNA/protein expression of IL-1β,IL-6,and TNF-α compared to the Control group,as well as intensified HIF-1α,IL-1β,and TNF-α fluores-cence signals.Conversely,the Force+LW6 group(mechanical force+30 μmol/L LW-6)exhibited a notable reduction in inflammatory cytokine expression levels and a weakening of HIF-1α,IL-1β,and TNF-α fluorescence signals com-pared to the Force group.Conclusion HIF-1α potentiates mechanical stress-induced inflammatory responses in hP-DLCs and may serve as a promising therapeutic target for mitigating orthodontic-associated periodontal inflammation.关键词
机械压力/人牙周膜细胞/缺氧诱导因子-1α/炎症应答/LW-6/炎症因子/骨改建/正畸治疗/碱性磷酸酶/信号通路Key words
mechanical stress/human periodontal ligament cells/hypoxia-inducible factor-1α/inflammatory response/LW-6/inflammatory factor/bone remodeling/orthodontic treatment/alkaline phosphatase/signal-ing pathway分类
医药卫生引用本文复制引用
王霏斐,郑程菊,陈芷芸,柳汀,王宇..HIF-1α促进机械压力下牙周膜细胞的炎症应答反应[J].口腔疾病防治,2025,33(9):732-743,12.基金项目
国家自然科学基金项目(82460199) （82460199）
贵州省科技计划项目(黔科合基础ZK[2022]一般384) （黔科合基础ZK[2022]一般384）
贵州医科大学国家自然科学基金培育项目(20NSP082) This study was supported by the grants from National Natural Science Foundation of China(No.82460199) （20NSP082）
Guizhou Provincial Science and Technology Projects(No.ZK2022YB384) （No.ZK2022YB384）
Guizhou Medical University Cultivation Project of Na-tional Natural Science Foundation(No.20NSP082). （No.20NSP082）
