Universidade de Taubaté - Unitau, School of Dentistry, Periodontics Research Division, Taubaté, SP, Brazil.
Universidade de São Paulo - USP, School of Dentistry, Departament of Periodontics, São Paulo, SP, Brazil.
Braz Oral Res. 2022 Apr 15;36:e048. doi: 10.1590/1807-3107bor-2022.vol36.0048. eCollection 2022.
PAR1 is a G-coupled protein receptor that regulates several cellular metabolism processes, including differentiation and proliferation of osteogenic and cementogenic related cells and our group previously demonstrated the regenerative potential of PAR1 in human periodontal ligament stem cells (hPDLSCs). In this study, we hypothesized that PAR1 regulates the cementogenic differentiation of hPDLSCs. Our goal was to identify the intracellular signaling pathway underlying PAR1 activation in hPDSLC differentiation. hPDLSCs were isolated using the explant technique. Cells were cultured in an osteogenic medium (OST) (α-MEM, 15% fetal bovine serum, L-glutamine, penicillin, streptomycin, amphotericin B, dexamethasone, and beta-glycerophosphate). The hPDLSCs were treated with a specific activator of PAR1 (PAR1 agonist) and blockers of the MAPK/ERK and PI3K pathways for 2 and 7 days. The gene expression of CEMP1 was assessed by RT-qPCR. The activation of PAR1 by its agonist peptide led to an increase in CEMP1 gene expression when compared with OST control. MAPK/ERK blockage abrogated the upregulation of CEMP1 gene expression induced by PAR1 agonist (p < 0.05). PI3K blockage did not affect the gene expression of CEMP1 at any experimental time (p > 0.05). We concluded that CEMP1 gene expression increased by PAR1 activation is MAPK/ERK-dependent and PI3K independent, suggesting that PAR1 may regulate cementogenetic differentiation of hPDLSCs.
PAR1 是一种 G 蛋白偶联受体，可调节多种细胞代谢过程，包括成骨细胞和造釉细胞的分化和增殖，我们之前的研究表明 PAR1 在前人牙周膜干细胞（hPDLSCs）中有再生潜能。在这项研究中，我们假设 PAR1 调节 hPDLSCs 的成牙骨质分化。我们的目的是鉴定 PAR1 激活在 hPDSLC 分化中涉及的细胞内信号通路。我们使用组织块培养法分离 hPDLSCs。将细胞培养在成骨培养基（OST）（α-MEM、15%胎牛血清、L-谷氨酰胺、青霉素、链霉素、两性霉素 B、地塞米松和β-甘油磷酸）中。用 PAR1 的特异性激活剂（PAR1 激动剂）和 MAPK/ERK 和 PI3K 通路的抑制剂处理 hPDLSCs 2 天和 7 天。通过 RT-qPCR 评估 CEMP1 的基因表达。与 OST 对照相比，PAR1 激动肽的激活导致 CEMP1 基因表达增加。MAPK/ERK 阻断消除了 PAR1 激动剂诱导的 CEMP1 基因表达的上调（p<0.05）。PI3K 阻断在任何实验时间都不会影响 CEMP1 的基因表达（p>0.05）。我们得出结论，PAR1 激活导致 CEMP1 基因表达增加是 MAPK/ERK 依赖性和 PI3K 非依赖性的，表明 PAR1 可能调节 hPDLSCs 的成牙骨质分化。
