Reciprocal role of PLAP‐1 in HIF‐1α‐mediated responses to hypoxia
Reciprocal role of PLAP‐1 in HIF‐1α‐mediated responses to hypoxia
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PLAP-1 在 HIF-1α 介导的缺氧反应中的相互作用
DOI：
10.1111/jre.12976
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发表时间：
2022
影响因子：
3.5
通讯作者：
Murakami Shinya
中科院分区：
文献类型：
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作者：
Takedachi Masahide;Yamamoto Satomi;Kawasaki Kohsuke;Shimomura Junpei;Murata Mari;Morimoto Chiaki;Hirai Asae;Kawakami Kazuma;Bhongsatiern Phan;Iwayama Tomoaki;Sawada Keigo;Yamada Satoru;Murakami Shinya
目的探讨缺氧诱导因子（HIF）-1 α活性与牙周膜相关蛋白-1（PLAP-1）表达在人牙周膜细胞（HPDL）中的相互调节作用。炎症和组织再生）通过激活HIF-1α。PLAP-1是一种优先在牙周膜中表达的细胞外基质蛋白，在HPDL的功能中发挥重要作用。虽然PLAP-1的表达已被证明在缺氧区域，参与PLAP-1的反应，缺氧尚未revealed.MethodsHPDLs培养在常氧（20%O2）或缺氧（1%O2）条件下，或不去铁胺甲磺酸盐（化学缺氧诱导剂）或chetomin（HIF信号抑制剂）。通过真实的实时逆转录聚合酶链反应和蛋白质印迹分析检测PLAP-1和HIF-1α的表达水平。荧光素酶报告基因HIF-1α活性的测定使用293 T细胞稳定转染缺氧反应元件（HRE）-含有荧光素酶载体的存在或不存在的重组PLAP-1或PLAP-1基因transfection. ResultsThe缺氧条件下培养的HPDL中PLAP-1的基因和蛋白表达水平升高。甲磺酸去铁胺治疗也增强了HPDL中PLAP-1的表达。缺氧诱导的PLAP-1表达在chetomin的存在下被显著抑制。PLAP-1抑制的HPDL在缺氧条件下培养期间显示细胞核中HIF-1α积累增加，但在存在重组PLAP-1的情况下则不然。在重组PLAP-1的存在下，缺氧诱导的293 T细胞的HRE活性以剂量依赖性方式被显著抑制。转染PLAP-1基因导致缺氧条件下培养过程中HRE活性显著降低。结论缺氧条件下PLAP-1表达通过HIF-1α激活而上调。此外，缺氧诱导的PLAP-1表达调节HIF-1α信号传导。
ObjectiveTo investigate the mutual regulation of hypoxia‐inducible factor (HIF)‐1α activity and periodontal ligament‐associated protein‐1 (PLAP‐1) expression in human periodontal ligament cells (HPDLs).BackgroundCellular responses to hypoxia regulate various biological events (e.g., inflammation and tissue regeneration) through activation of HIF‐1α. PLAP‐1, an extracellular matrix protein preferentially expressed in the periodontal ligament, plays important roles in the functions of HPDLs. Although PLAP‐1 expression has been demonstrated in hypoxic regions, the involvement of PLAP‐1 in responses to hypoxia has not been revealed.MethodsHPDLs were cultured under normoxic (20% O2) or hypoxic (1% O2) conditions with or without deferoxamine mesylate (chemical hypoxia inducer) or chetomin (HIF signaling inhibitor). Expression levels of PLAP‐1 and HIF‐1α were examined by real‐time reverse transcription‐polymerase chain reaction and western blot analysis. Luciferase reporter assays of HIF‐1α activity were performed using 293T cells stably transfected with a hypoxia response element (HRE)‐containing luciferase vector in the presence or absence of recombinant PLAP‐1 or PLAP‐1 gene transfection.ResultsCultivation under hypoxic conditions elevated the gene and protein expression levels of PLAP‐1 in HPDLs. Deferoxamine mesylate treatment also enhanced PLAP‐1 expression in HPDLs. Hypoxia‐induced PLAP‐1 expression was significantly suppressed in the presence of chetomin. PLAP‐1‐suppressed HPDLs showed increased HIF‐1α accumulation in the nucleus during culture under hypoxic conditions, but not in the presence of recombinant PLAP‐1. In the presence of recombinant PLAP‐1, hypoxia‐induced HRE activity of 293T cells was significantly suppressed in a dose‐dependent manner. Transfection of the PLAP‐1 gene resulted in a significant reduction of HRE activity during culture under hypoxic conditions.ConclusionPLAP‐1 expression is upregulated under hypoxic conditions through HIF‐1α activation. Moreover, hypoxia‐induced PLAP‐1 expression regulates HIF‐1α signaling.