草药忍冬藤通过调节 NF-κB 和 MAPK 通路抑制 RANKL 诱导的破骨细胞分化和骨吸收活性。

Herbal medicine Ninjinyoeito inhibits RANKL-induced osteoclast differentiation and bone resorption activity by regulating NF-κB and MAPK pathway.

机构信息

Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan.

Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan; Department of Oral and Maxillofacial Surgery, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, 700-8525, Japan.

出版信息

J Oral Biosci. 2024 Dec;66(4):49-57. doi: 10.1016/j.job.2024.09.007. Epub 2024 Oct 2.

DOI:10.1016/j.job.2024.09.007

PMID:39366652

Abstract

OBJECTIVES

Osteoporosis is a systemic bone metabolism disorder characterized by decreased bone mass and strength. Osteoclasts (OCs) are giant multinucleated cells that regulate bone homeostasis by degrading bone matrix. Excessive OC differentiation and activity can lead to serious bone metabolic disorders including osteoporosis. Current treatments, including antiresorptive drugs, exert considerable adverse effects, including jaw osteonecrosis. Herbal medicines, such as Ninjinyoeito (NYT), may also offer efficacy, but with fewer adverse effects. In this study, we investigated NYT's effects on osteoclastogenesis.

METHODS

Tartrate-resistant acid phosphatase (TRAP) staining and bone resorption assays were performed to examine NYT's effects on OC differentiation and function. OC-related gene expression at mRNA and protein levels was investigated to confirm NYT's inhibitory action against osteoclastogenesis. We also demonstrated involvement of signaling pathways mediated by IκBα and mitogen-activated protein kinases (MAPK) [extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38] and showed nuclear translocation of nuclear factor of activated T-cell cytoplasmic 1 (NFATc1) and nuclear factor kappa B (NF-κB) p65 during osteoclastogenesis.

RESULTS

TRAP staining and bone resorption assays confirmed that NYT significantly inhibited OC differentiation and function. Western blot and RT-PCR results showed that NYT ameliorated osteoclastogenesis by suppressing mRNA and protein level expression of OC-related genes. Moreover, blots and immunocytochemistry (ICC) data clarified that NYT abrogates signaling pathways mediated by IκBα and MAPK (ERK, JNK, p38), and demonstrated nuclear translocation of NFATc1 and NF-κB p65 during OC differentiation.

CONCLUSIONS

These findings suggest NYT is an alternative therapeutic candidate for treating osteoporosis.

摘要

目的

骨质疏松症是一种以骨量减少和骨强度降低为特征的系统性骨代谢紊乱。破骨细胞（OC）是一种巨大的多核细胞，通过降解骨基质来调节骨稳态。OC 分化和活性的过度增加可导致严重的骨骼代谢紊乱，包括骨质疏松症。目前的治疗方法，包括抗吸收药物，会产生相当大的不良反应，包括颌骨坏死。草药，如 Ninjinyoeito（NYT），也可能具有疗效，但不良反应较少。在这项研究中，我们研究了 NYT 对破骨细胞生成的影响。

方法

采用抗酒石酸酸性磷酸酶（TRAP）染色和骨吸收试验来检测 NYT 对 OC 分化和功能的影响。通过检测 OC 相关基因的 mRNA 和蛋白水平的表达来证实 NYT 对破骨细胞生成的抑制作用。我们还证明了 IκBα 和丝裂原活化蛋白激酶（MAPK）[细胞外信号调节激酶（ERK）、c-Jun N-末端激酶（JNK）和 p38]介导的信号通路的参与，并显示了在破骨细胞生成过程中核因子活化 T 细胞胞浆 1（NFATc1）和核因子 kappa B（NF-κB）p65 的核转位。

结果

TRAP 染色和骨吸收试验证实 NYT 显著抑制 OC 的分化和功能。Western blot 和 RT-PCR 结果表明，NYT 通过抑制 OC 相关基因的 mRNA 和蛋白表达水平来改善破骨细胞生成。此外，印迹和免疫细胞化学（ICC）数据表明，NYT 阻断了 IκBα 和 MAPK（ERK、JNK、p38）介导的信号通路，并证明了 NFATc1 和 NF-κB p65 在 OC 分化过程中的核转位。