Dragon blood resin ameliorates steroid-induced osteonecrosis of femoral head through osteoclastic pathways

Yuhao Liu; Liang Mo; Hongduo Lu; Yangwenxiang Wei; Jiahao Zhang; Samuel Bennett; Jiake Xu; Chi Zhou; Bin Fang; Zhenqiu Chen

doi:10.3389/fcell.2023.1202888

Dragon blood resin ameliorates steroid-induced osteonecrosis of femoral head through osteoclastic pathways

Front Cell Dev Biol. 2023 Aug 22:11:1202888. doi: 10.3389/fcell.2023.1202888. eCollection 2023.

Authors

Yuhao Liu^#¹, Liang Mo^#¹, Hongduo Lu¹, Yangwenxiang Wei¹, Jiahao Zhang¹, Samuel Bennett², Jiake Xu^{2

3}, Chi Zhou¹, Bin Fang¹, Zhenqiu Chen¹

Affiliations

¹ The First Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou, China.
² School of Biomedical Sciences, University of Western Australia, Perth, WA, Australia.
³ Shenzhen institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.

^# Contributed equally.

Abstract

Objective: Dragon's Blood resin (DBR) is a traditional medicinal substance renowned for its diverse pharmacological effects, which consists of potent anti-inflammatory, antioxidant and angiogenic properties. This study aimed to elucidate its therapeutic mechanism in alleviating steroid-induced osteonecrosis of the femoral head (SIONFH). Methods: Techniques such as SPR and LC-MS were employed to identify and analyze the target proteins of DBR in bone marrow macrophages (BMMs). In vitro, BMMs were treated with RANKL and DBR, and TRAcP staining and actin belt staining were utilized to assess osteoclast activity. The inhibitory effects and underlying mechanisms of DBR on osteoclastogenesis and reactive oxygen species (ROS) generation were determined using real-time PCR, western blotting and immunofluorescence staining. An in vivo SIONFH rat model was set up to assess the curative impacts of DBR using micro-CT scanning and pathological staining. Results: Bioinformatic tools revealed a pivotal role of osteoclast differentiation in SIONFH. Proteomic analysis identified 164 proteins binding in BMMs. In vitro assessments demonstrated that DBR hindered osteoclastogenesis by modulating the expression of specific genes and proteins, along with antioxidant proteins including TRX1 and Glutathione Reductase. Notably, the resin effectively inhibited the expression of crucial proteins, such as the phosphorylation of JNK and the nuclear localization of p65 within the TRAF6/JNK and NFκB signaling pathways. In vivo experiments further confirmed that DBR mitigated the onset of SIONFH in rats by curbing osteoclast and ROS activities. Conclusion: These findings underscore the potential of Dragon's Blood as an effective administration for early-stage SIONFH, shedding light on its therapeutic influence on ROS-mediated osteoclastic signaling pathways.

Keywords: Dracaena cochinchinensis; Dragon’s blood resin; osteoclast; osteonecrosis of the femoral head; reactive oxygen species.

Grants and funding

This research was supported by Natural Science Foundation of Guangdong Province (Grant No. 2021A1515011484), Guangzhou Science and Technology Bureau (Grant No. 202102020930 and 202201020314), National Natural Science Foundation of China (Grant No. 82104883), Traditional Chinese Medicine Bureau of Guangdong Province (Grant Nos. 20221136 and 20231162) and Double First-rate Discipline and High-level University Construction Projects of Guangzhou University of Chinese Medicine (Grant No. 2021xk46). JX was supported by the Australian National Health and Medical Research Council (Grant Nos. APP1107828, APP1127156, and APP1163933).