破骨细胞的过度活动导致许多骨相关疾病,如类风湿性关节炎和骨质疏松症。Agrimopol(AGR),酚类化合物,起源于AgrimoniapilosaLedeb.在先前的研究中,据报道,AGR具有杀血吸虫和杀分枝杆菌活性。然而,没有报告涵盖其抗破骨细胞生成特性。在这项研究中,我们发现AGR抑制RANKL诱导的破骨细胞生成,骨吸收,F-肌动蛋白环的形成,和破骨细胞相关基因如CTSK的mRNA表达,陷阱,MMP-9和ATP6v0d2在体外。此外,AGR抑制RANKL诱导的c-Fos和NFATc1表达。然而,AGR处理不影响RANKL刺激的BMM中的NF-κB激活和MAPK磷酸化,这表明AGR可能不会影响NF-κB和MAPK信号传导介导的NFATc1的初始表达。我们的结果进一步表明,AGR并没有改变GSK3β的磷酸化水平和钙调磷酸酶的表达,这表明AGR治疗可能不会干扰GSK3β和钙调磷酸酶介导的NFATc1的磷酸化和去磷酸化,分别。B淋巴细胞诱导成熟蛋白-1(Blimp1),它被认为是破骨细胞生成的负调节因子的转录抑制因子,在AGR的存在下明显减弱,导致B细胞淋巴瘤6(Bcl-6)的表达增强。同时,通过siRNA在BMM中敲低Blimp1强烈增强Bcl6的表达并减少RANKL对NFATc1的诱导。这些发现表明,AGR通过Blimp1-Bcl-6信号介导的NFATc1及其靶基因的调节来抑制RANKL诱导的破骨细胞分化。与这些体外结果一致,AGR通过抑制过度的破骨细胞活性和减轻LPS诱导的骨破坏,在LPS诱导的骨丢失的体内小鼠模型中表现出保护性影响。因此,这些结果确定AGR可被视为一种潜在的骨溶解病治疗剂.
Excessive activity of osteoclasts causes many bone-related diseases, such as rheumatoid arthritis and osteoporosis. Agrimophol (AGR), a phenolic compound, originated from Agrimonia pilosa Ledeb. In prior studies, AGR is reported to possess schistosomicidal and mycobactericidal activities. However, no reports covered its anti-osteoclastogenesis characteristic. In this study, we found that AGR inhibited RANKL-induced osteoclastogenesis, bone-resorption, F-actin ring formation, and the mRNA expression of osteoclast-associated genes such as CTSK, TRAP, MMP-9, and ATP6v0d2 in vitro. In addition, AGR suppressed RANKL-induced expression of c-Fos and NFATc1. However, AGR treatment did not affect NF-κB activation and MAPKs phosphorylation in RANKL-stimulated BMMs, which implicated that AGR might not influence the initial expression of NFATc1 mediated by NF-κB and MAPKs signaling. Our results further indicated that AGR did not alter phosphorylation levels of GSK3β and the expression of calcineurin, which implicated that AGR treatment might not interfere with phosphorylation and de-phosphorylation of NFATc1 mediated by GSK3β and calcineurin, respectively. B-lymphocyte-induced maturation protein-1 (
Blimp1), which was regarded as a transcriptional repressor of negative regulators of osteoclastogenesis, was markedly attenuated in the presence of AGR, leading to the enhanced expression of B-cell lymphoma 6 (Bcl-6). Meanwhile,
Blimp1 knockdown in BMMs by siRNA strongly enhanced the expression of Bcl6 and reduced NFATc1 induction by RANKL. These findings suggested that AGR inhibited RANKL-induced osteoclast differentiation through
Blimp1-Bcl-6 signaling mediated modulation of NFATc1 and its target genes. Consistent with these in vitro results, AGR exhibited a protective influence in an in vivo mouse model of LPS-induced bone loss by suppressing excessive osteoclast activity and attenuating LPS-induced bone destruction. Hence, these results identified that AGR could be considered as a potential therapeutic agent against bone lysis disease.