Department of Periodontology, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, No. 44-1 Wenhua Road West, 250012, Jinan, People's Republic of China.
Clin Oral Investig. 2021 Feb;25(2):581-592. doi: 10.1007/s00784-020-03498-9. Epub 2020 Aug 12.
To determine the role of angiogenic factor with G-patch and FHA domain 1 (AGGF1) in inflammatory response of human dental pulp cells (DPCs) and the underneath mechanism and to explore its role in angiogenesis.
The expression of AGGF-1 in human healthy and inflammatory pulp tissues was detected by immunohistochemistry. RT-qPCR and Western blot were used to evaluate the expression of AGGF1 in DPCs stimulated by lipopolysaccharide (LPS). After AGGF1 was knocked down, the expression of LPS-induced inflammatory cytokines in DPCs was quantified by RT-qPCR and ELISA. Immunofluorescence and Western blot were used to assess the activation of NF-κB signaling. Inflammatory cytokines were detected by RT-qPCR and ELISA in DPCs pretreated with NF-κB pathway inhibitors before LPS stimulation, and then the effect of AGGF1 on angiogenesis was also evaluated.
AGGF1 expression increased in inflammatory dental pulp tissues. In DPCs stimulated by LPS, AGGF1 was upregulated in a dose-dependent manner (P < 0.05). In AGGF1 knockdown cells, the expression of IL-6, IL-8, and monocyte chemoattractant protein-1 (MCP-1/CCL-2) increased by LPS stimulation (P < 0.001). Nuclear translocation of p65 was promoted, and the addition of NF-κB inhibitors inhibited the expression of inflammatory factors. Meanwhile, knockdown of AGGF1 inhibited vascularization.
AGGF1 inhibited the synthesis of inflammatory cytokines through NF-κB signaling pathway and promoted the angiogenesis of DPCs.
This study might shed light in the treatment of pulpitis and regeneration of dental pulp tissues; however, more clinical trials are required to validate these findings.
确定含 G- 补丁和 FHA 结构域 1（AGGF1）的血管生成因子在人牙髓细胞（DPC）炎症反应中的作用及其潜在机制，并探讨其在血管生成中的作用。
采用免疫组织化学法检测人健康和炎症牙髓组织中 AGGF-1 的表达。用脂多糖（LPS）刺激 DPC 后，通过 RT-qPCR 和 Western blot 评估 AGGF1 的表达。敲低 AGGF1 后，通过 RT-qPCR 和 ELISA 定量检测 LPS 诱导的 DPC 中炎症细胞因子的表达。用免疫荧光和 Western blot 评估 NF-κB 信号通路的激活。在 LPS 刺激前，用 NF-κB 通路抑制剂预处理 DPC，检测 DPC 中炎症细胞因子的表达，然后评估 AGGF1 对血管生成的影响。
炎症性牙髓组织中 AGGF1 表达增加。LPS 刺激的 DPC 中，AGGF1 呈剂量依赖性上调（P < 0.05）。在 AGGF1 敲低细胞中，LPS 刺激后 IL-6、IL-8 和单核细胞趋化蛋白-1（MCP-1/CCL-2）的表达增加（P < 0.001）。p65 核转位增加，NF-κB 抑制剂的加入抑制了炎症因子的表达。同时，敲低 AGGF1 抑制了血管生成。
AGGF1 通过 NF-κB 信号通路抑制炎症细胞因子的合成，促进 DPC 的血管生成。
本研究可能为牙髓炎的治疗和牙髓组织再生提供思路；然而，还需要更多的临床试验来验证这些发现。
