Department of Orthopedics, School of Medicine, China Medical University Hospital and China Medical University, Taichung, Taiwan.
Department of Chemistry, National Changhua University of Education, Changhua, Taiwan.
Eur J Pharmacol. 2018 Aug 5;832:104-113. doi: 10.1016/j.ejphar.2018.05.024. Epub 2018 May 18.
Previous studies have indicated that paeonol inhibits RANKL-induced osteoclastogenesis by inhibiting the ERK, p38, and NF-κB pathway. We modified paeonol to form a new compound, YPH-PA3, and found that it promoted osteoclastogenesis rather than inhibited it the way paeonol does. The aim of this study is to investigate the mechanisms involved in YPH-PA3-promoted osteoclastogenesis. YPH-PA3-promoted differentiation of RAW264.7 cells (human monocytes) into osteoclasts is activated through ERK/p38/JNK phosphorylation, affecting c-FOS, NF-κB, and NFATc2. Real-time quantitative PCR and western blot revealed an increased expression of autophagy-related markers during YPH-PA3-induced osteoclastogenesis. We also demonstrated the relationship between p62/LC3 localization and F-actin ring formation by double-labeling immunofluorescence. Knockdown of p62 small-interfering RNA (siRNA) attenuated YPH-PA3-induced expression of autophagy-related genes. Our study results indicated that p62 may play a role in YPH-PA3-induced autophagy and osteoclastogenesis, which may help to develop a novel therapeutic strategy against osteoclastogenesis-related diseases.
先前的研究表明，丹皮酚通过抑制 ERK、p38 和 NF-κB 通路抑制 RANKL 诱导的破骨细胞分化。我们对丹皮酚进行了修饰，形成了一种新的化合物 YPH-PA3，并发现它促进了破骨细胞的分化，而不是像丹皮酚那样抑制破骨细胞的分化。本研究旨在探讨 YPH-PA3 促进破骨细胞分化的机制。YPH-PA3 通过 ERK/p38/JNK 磷酸化激活 RAW264.7 细胞（人单核细胞）向破骨细胞的分化，影响 c-FOS、NF-κB 和 NFATc2。实时定量 PCR 和 Western blot 显示，在 YPH-PA3 诱导的破骨细胞分化过程中，自噬相关标记物的表达增加。我们还通过双标记免疫荧光显示了 p62/LC3 定位与 F-肌动蛋白环形成之间的关系。p62 小干扰 RNA (siRNA) 的敲低减弱了 YPH-PA3 诱导的自噬相关基因的表达。我们的研究结果表明，p62 可能在 YPH-PA3 诱导的自噬和破骨细胞分化中发挥作用，这可能有助于开发针对破骨细胞分化相关疾病的新的治疗策略。
