颗粒蛋白前体通过肿瘤坏死因子受体促进人牙周膜干细胞的成骨分化，以抑制 TNF-α 敏化的 NF-κB 并激活 ERK/JNK 信号通路。
Progranulin promotes osteogenic differentiation of human periodontal ligament stem cells via tumor necrosis factor receptors to inhibit TNF-α sensitized NF-kB and activate ERK/JNK signaling.
机构信息
Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of Stomatology, Shandong University, Jinan, China.
Department of Periodontology, School of Stomatology, Shandong University, Jinan, China.
出版信息
J Periodontal Res. 2020 Jun;55(3):363-373. doi: 10.1111/jre.12720. Epub 2019 Dec 19.
OBJECTIVE
To investigate the molecular mechanism of Progranulin (PGRN) in promoting osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) in inflammatory environment.
BACKGROUND
Progranulin is an antagonist of tumor necrosis factor (TNF) receptors (TNFRs) and is known to promote inflammatory periodontal bone defect regeneration.
METHODS
TNFR1- and TNFR2-silenced hPDLSCs designed as hPDLSCs-sh-TNFR1 and hPDLSCs-sh-TNFR2 were cultured with osteoinductive medium containing TNF-α and (or) PGRN. Immunofluorescence, quantitative real-time PCR, and western blot were used to, respectively, detect expressions of TNFR1\TNFR2 and osteogenic differentiation markers as well as phosphorylation level in NF-κB\MAPK-related pathways.
RESULTS
Immunofluorescence and real-time PCR showed that TNFR1 and TNFR2 positively expressed in hPDLSCs. TNF-α stimulation could significantly decrease the expressions of ALP and RUNX2 in hPDLSCs, whereas PGRN treatment could significantly enhance their expressions, and reverse TNF-α-mediated expression suppression of ALP and RUNX2 in hPDLSCs. In hPDLSCs-sh-TNFR1, TNF-α mediated osteogenic inhibition decreased, but both TNF-α + PGRN and alone PGRN significantly promoted expression of ALP and RUNX2. PGRN significantly enhanced expression of P-ERK1/2 and P-JNK, while corresponding inhibitors eliminated PGRN-stimulated osteogenic differentiation. In hPDLSCs-sh-TNFR2, no significant difference existed in osteogenic markers and P-JNK expression between the PGRN group and the control group. However, PGRN still activated P-ERK1/2 expression. Besides, PGRN antagonized TNF-α-enhanced NF-κB P65 expression.
CONCLUSION
Progranulin promotes osteogenic differentiation of hPDLSCs via TNFR1 to inhibit TNF-α-sensitized NF-κB and via TNFR2 to activate JNK signaling. The mechanism by which PGRN activates ERK signaling remains to be explored.
目的
探讨颗粒体蛋白聚糖（PGRN）在炎症环境下促进人牙周膜干细胞（hPDLSCs）成骨分化的分子机制。
背景
颗粒体蛋白聚糖是肿瘤坏死因子（TNF）受体（TNFRs）的拮抗剂，已知其可促进炎症性牙周骨缺损再生。
方法
设计 TNFR1 和 TNFR2 沉默的 hPDLSCs 作为 hPDLSCs-sh-TNFR1 和 hPDLSCs-sh-TNFR2，在含有 TNF-α 和（或）PGRN 的成骨诱导培养基中培养。免疫荧光、实时定量 PCR 和 Western blot 分别用于检测 TNFR1/TNFR2 和成骨分化标志物的表达以及 NF-κB/MAPK 相关通路的磷酸化水平。
结果
免疫荧光和实时 PCR 显示 TNFR1 和 TNFR2 在 hPDLSCs 中呈阳性表达。TNF-α 刺激可显著降低 hPDLSCs 中 ALP 和 RUNX2 的表达，而 PGRN 处理可显著增强其表达，并逆转 TNF-α 介导的 hPDLSCs 中 ALP 和 RUNX2 的表达抑制。在 hPDLSCs-sh-TNFR1 中，TNF-α 介导的成骨抑制作用减弱，但 TNF-α+PGRN 和单独 PGRN 均显著促进 ALP 和 RUNX2 的表达。PGRN 显著增强了 P-ERK1/2 和 P-JNK 的表达，而相应的抑制剂消除了 PGRN 刺激的成骨分化。在 hPDLSCs-sh-TNFR2 中，PGRN 组与对照组之间成骨标志物和 P-JNK 表达无显著差异。然而，PGRN 仍能激活 P-ERK1/2 的表达。此外，PGRN 拮抗 TNF-α 增强的 NF-κB P65 表达。
结论
PGRN 通过 TNFR1 促进 hPDLSCs 的成骨分化，抑制 TNF-α 敏化的 NF-κB，通过 TNFR2 激活 JNK 信号通路。PGRN 激活 ERK 信号的机制仍有待探索。
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