Department of Pharmacology, Guangdong Key Laboratory for Research and Development of Natural Drugs, the affiliated hospital, Department of Physiology, Guangdong Medical University, Zhanjiang 524023, China.
Department of Pharmacology, Guangdong Key Laboratory for Research and Development of Natural Drugs, the affiliated hospital, Department of Physiology, Guangdong Medical University, Zhanjiang 524023, China.
Phytomedicine. 2023 Jul 25;116:154875. doi: 10.1016/j.phymed.2023.154875. Epub 2023 May 22.
Osteoporosis (OP) is considered as one of the major comorbidities of rheumatoid arthritis (RA), and is responsible for fragility fracture. However, there is currently no effective treatment for RA complicated with OP. Tubson-2 decoction (TBD), a Mongolian medicine also known as Erwei Duzhong Decoction, has been shown to exert a preventive effect on post-menopausal osteoporosis (PMOP). The preventive effects of TBD on RA-induced OP, as well as the bioactive compound responsible and the underlying mechanisms, remain to be elucidated.
To explore the effects of TBD on RA-induced OP in vivo, and to elucidate the mechanism of isochlorogenic acid A (ICA), the effective component of TBD, in vitro.
To evaluate the anti-arthritic and anti-osteoporotic effects of TBD, we conducted H&E straining and safranine O/fast green, TEM, immunohistochemistry (IHC), bone histomorphometry, micro-CT imaging, and biomechanical testing in collagen induced arthritis (CIA) rats. The active ingredient in TBD was identified using network pharmacology and molecular docking. The identification was supported by in vivo IHC assay, and further confirmed using qRT-PCR, Western blot, and SEM analysis in TNF-α-treated MH7A cells and/or in LPS-exposed RAW264.7 cells.
Oral administration of TBD attenuated the severity of arthritis and osteopenia as well as poor bone quality, in CIA rats. Additionally, TBD and the positive control, tripterygium glycosides (TG), exhibited similar effects in reducing inflammation in both the synovium and ankle joint. They also were both effective in improving bone loss, microarchitecture, and overall bone quality. TBD reduced the expression of MMP13, IL-17, and p-JNK protein in the synovium of CIA rats. ICA, which was screened, suppressed TNF-α or LPS-triggered inflammatory responses via down-regulating IL-17 signaling, involving in MMP13, IL-1β, IL-23, and IL-17, and the MAPK pathway including p-ERK, p-JNK, and p-P38, both in MH7A cells and in RAW264.7 cells. Furthermore, ICA prevented osteoclasts from differentiating and bone resoprtion in a dose-dependent manner in vitro.
This study provides the first evidence that TBD exerts intervening effects on RA-induced OP, possibly through the downregulation of the IL-17/MAPK signaling pathway by ICA. The findings of our study provides valuable insights for further research in this area.
骨质疏松症（OP）被认为是类风湿关节炎（RA）的主要合并症之一，可导致脆性骨折。然而，目前尚无有效的方法治疗 RA 合并 OP。蒙药图荪-2 汤（TBD），也被称为二味杜仲汤，已被证明对绝经后骨质疏松症（PMOP）具有预防作用。TBD 对 RA 诱导的 OP 的预防作用，以及负责的生物活性化合物及其潜在机制仍有待阐明。
探讨 TBD 对体内 RA 诱导的 OP 的影响，并阐明 TBD 的有效成分异绿原酸 A（ICA）在体外的作用机制。
为了评估 TBD 的抗关节炎和抗骨质疏松作用，我们对胶原诱导性关节炎（CIA）大鼠进行了 H&E 染色和番红 O/固绿、TEM、免疫组织化学（IHC）、骨组织形态计量学、微 CT 成像和生物力学测试。使用网络药理学和分子对接鉴定 TBD 的活性成分。通过体内 IHC 测定对鉴定进行支持，并进一步通过 TNF-α 处理的 MH7A 细胞和/或 LPS 暴露的 RAW264.7 细胞中的 qRT-PCR、Western blot 和 SEM 分析进行确认。
TBD 口服给药可减轻 CIA 大鼠关节炎和骨质疏松症的严重程度以及骨质量差。此外，TBD 和阳性对照雷公藤多苷（TG）在减轻滑膜和踝关节炎症方面均具有相似的作用。它们还能有效改善骨质流失、微结构和整体骨质量。TBD 降低了 CIA 大鼠滑膜中 MMP13、IL-17 和 p-JNK 蛋白的表达。筛选出的 ICA 通过下调 IL-17 信号通路抑制 TNF-α 或 LPS 触发的炎症反应，涉及 MMP13、IL-1β、IL-23 和 IL-17 以及 MAPK 通路包括 p-ERK、p-JNK 和 p-P38，在 MH7A 细胞和 RAW264.7 细胞中均如此。此外，ICA 还可以在体外以剂量依赖的方式阻止破骨细胞分化和骨吸收。
本研究首次提供了 TBD 对 RA 诱导的 OP 具有干预作用的证据，这可能是通过 ICA 下调 IL-17/MAPK 信号通路实现的。我们的研究结果为该领域的进一步研究提供了有价值的见解。
