Kim Yun Sook, Otgonsuren Munkh-Ochir
Department of Anatomy and Neurobiology, School of Dentistry, Kyungpook National University, Daegu, the Republic of Korea.
Department of Anatomy and Neurobiology, School of Dentistry, Kyungpook National University, Daegu, the Republic of Korea.
Arch Oral Biol. 2023 Nov;155:105800. doi: 10.1016/j.archoralbio.2023.105800. Epub 2023 Sep 1.
To investigate whether transient receptor potential ankyrin 1 (TRPA1) and transient receptor potential melastatin 8 (TRPM8) have a function in responding to environmental stimuli in human odontoblast-like cells (hOLCs). Additionally, to explore whether activation of TRPA1 and TRPM8 in hOLCs participates in the regulation of the inflammatory process.
Changes in gene and protein expression levels of TRPA1 and TRPM8 in cultured hOLCs following lipopolysaccharide (LPS) stimulation, which mimics inflammation, were examined using quantitative reverse transcription-polymerase chain reaction and western blot analysis. Furthermore, we compared the expression profiles of 80 cytokines between LPS- and vehicle-treated hOLCs and investigated how the production of highly increased cytokines in LPS-treated hOLCs was affected by the pharmacological inhibition of TRPA1 and TRPM8.
The expression of TRPA1 and TRPM8 in hOLCs was observed and their mRNAs and proteins were upregulated in hOLCs after LPS treatment. Moreover, cytokine antibody assays revealed that monocyte chemoattractant protein-1 (MCP-1, CCL2), growth-regulated protein α (GROα, CXCL1), interleukin-6 (IL-6), and IL-8 (CXCL8) were significantly upregulated by LPS. The pharmacological inhibition of TRPA1 (HC-030031) during LPS treatment attenuated the expression of CCL2, CXCL1, and IL-8, whereas the pharmacological inhibition of TRPM8 (PF05105679) suppressed the expression of CCL2, CXCL1, and IL-8 as well as IL-6.
These results indicate that hOLCs express TRPA1 and TRPM8, which are upregulated during inflammation. In addition to being sensors of potentially harmful stimuli, TRPA1 and TRPM8 in hOLCs play important roles in regulating inflammatory responses.
研究瞬时受体电位锚蛋白1(TRPA1)和瞬时受体电位 melastatin 8(TRPM8)在人成牙本质细胞样细胞(hOLCs)对环境刺激的反应中是否具有功能。此外,探讨hOLCs中TRPA1和TRPM8的激活是否参与炎症过程的调节。
使用定量逆转录-聚合酶链反应和蛋白质印迹分析,检测脂多糖(LPS)刺激(模拟炎症)后培养的hOLCs中TRPA1和TRPM8的基因和蛋白表达水平变化。此外,我们比较了LPS处理组和溶剂处理组hOLCs之间80种细胞因子的表达谱,并研究了TRPA1和TRPM8的药理学抑制如何影响LPS处理的hOLCs中高度增加的细胞因子的产生。
观察到hOLCs中TRPA1和TRPM8的表达,LPS处理后hOLCs中的mRNA和蛋白上调。此外,细胞因子抗体检测显示,LPS显著上调单核细胞趋化蛋白-1(MCP-1,CCL2)、生长调节蛋白α(GROα,CXCL1)、白细胞介素-6(IL-6)和IL-8(CXCL8)。LPS处理期间TRPA1的药理学抑制(HC-030031)减弱了CCL2、CXCL1和IL-8的表达,而TRPM8的药理学抑制(PF05105679)抑制了CCL2、CXCL1和IL-8以及IL-6的表达。
这些结果表明hOLCs表达TRPA1和TRPM8,它们在炎症期间上调。除了作为潜在有害刺激的传感器外,hOLCs中的TRPA1和TRPM8在调节炎症反应中起重要作用。