Manokawinchoke J, Pimkhaokhum A, Everts V, Pavasant P
Mineralized Tissue Research Unit, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
J Periodontal Res. 2014 Dec;49(6):777-84. doi: 10.1111/jre.12162. Epub 2014 Jan 11.
Prostaglandin E2 (PGE2) has been shown to be able to influence both bone formation and resorption. The purpose of this study was to investigate the effect of PGE2 on the osteogenic differentiation of human periodontal ligament (HPDL) cells.
HPDL cells were cultured with 0.001-1 μm PGE2 in osteogenic medium. In-vitro mineral deposition was determined by Alizarin Red S staining, and gene expression was determined by real-time PCR.
PGE2 inhibited in-vitro mineral deposition by HPDL cells in a dose-dependent manner. PCR analyses showed that PGE2 upregulated the expression of Runt-related transcription factor 2 (RUNX2), but had no effect on osteocalcin expression. Upregulation of TWIST-related protein1 (TWIST1), a functional antagonist of RUNX2, was also observed. In addition, increased levels of RUNX2 and TWIST1 proteins, induced by PGE2, were detected by western blot analysis. Using a chemical activator of E prostanoid (EP) receptors as well as small interfering RNA against an EP receptor, it was shown that PGE2 regulated RUNX2 and TWIST1 via the EP2 receptor. The role of protein kinase A in the inductive effect of PGE2 was also demonstrated.
The results of this study revealed that PGE2 modulates the osteogenic differentiation of HPDL cells via regulating the expression of RUNX2 and TWIST1. The results suggest a possible role for PGE2 in regulating the homeostasis of periodontal ligament tissue.
前列腺素E2(PGE2)已被证明能够影响骨形成和骨吸收。本研究的目的是探讨PGE2对人牙周膜(HPDL)细胞成骨分化的影响。
将HPDL细胞在成骨培养基中与0.001 - 1μm的PGE2一起培养。通过茜素红S染色测定体外矿化沉积,并通过实时PCR测定基因表达。
PGE2以剂量依赖性方式抑制HPDL细胞的体外矿化沉积。PCR分析表明,PGE2上调了Runt相关转录因子2(RUNX2)的表达,但对骨钙素表达没有影响。还观察到RUNX2的功能性拮抗剂TWIST相关蛋白1(TWIST1)上调。此外,通过蛋白质印迹分析检测到PGE2诱导的RUNX2和TWIST1蛋白水平升高。使用前列腺素E(EP)受体的化学激活剂以及针对EP受体的小干扰RNA,表明PGE2通过EP2受体调节RUNX2和TWIST1。还证明了蛋白激酶A在PGE2诱导作用中的作用。
本研究结果表明,PGE2通过调节RUNX2和TWIST1的表达来调节HPDL细胞的成骨分化。结果提示PGE2在调节牙周膜组织稳态中可能发挥作用。