Liu Yuhao, Mo Liang, Lu Hongduo, Wei Yangwenxiang, Zhang Jiahao, Bennett Samuel, Xu Jiake, Zhou Chi, Fang Bin, Chen Zhenqiu
The First Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou, China.
School of Biomedical Sciences, University of Western Australia, Perth, WA, Australia.
Front Cell Dev Biol. 2023 Aug 22;11:1202888. doi: 10.3389/fcell.2023.1202888. eCollection 2023.
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). Techniques such as SPR and LC-MS were employed to identify and analyze the target proteins of DBR in bone marrow macrophages (BMMs). , 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 SIONFH rat model was set up to assess the curative impacts of DBR using micro-CT scanning and pathological staining. Bioinformatic tools revealed a pivotal role of osteoclast differentiation in SIONFH. Proteomic analysis identified 164 proteins binding in BMMs. 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. experiments further confirmed that DBR mitigated the onset of SIONFH in rats by curbing osteoclast and ROS activities. 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.
血竭树脂(DBR)是一种传统药物,以其多样的药理作用而闻名,具有强大的抗炎、抗氧化和血管生成特性。本研究旨在阐明其在缓解类固醇诱导的股骨头坏死(SIONFH)中的治疗机制。采用表面等离子体共振(SPR)和液相色谱-质谱联用(LC-MS)等技术来鉴定和分析血竭树脂在骨髓巨噬细胞(BMMs)中的靶蛋白。用核因子κB受体活化因子配体(RANKL)和血竭树脂处理BMMs,并利用抗酒石酸酸性磷酸酶(TRAcP)染色和肌动蛋白带染色来评估破骨细胞活性。使用实时聚合酶链反应(PCR)、蛋白质免疫印迹法和免疫荧光染色来确定血竭树脂对破骨细胞生成和活性氧(ROS)产生的抑制作用及其潜在机制。建立SIONFH大鼠模型,通过显微计算机断层扫描(micro-CT)和病理染色来评估血竭树脂的治疗效果。生物信息学工具揭示了破骨细胞分化在SIONFH中的关键作用。蛋白质组学分析鉴定出164种在BMMs中结合的蛋白质。评估表明,血竭树脂通过调节特定基因和蛋白质的表达,以及包括硫氧还蛋白1(TRX1)和谷胱甘肽还原酶在内的抗氧化蛋白,来阻碍破骨细胞生成。值得注意的是,该树脂有效抑制了关键蛋白的表达,如TRAF6/JNK和NFκB信号通路中JNK的磷酸化和p65的核定位。实验进一步证实,血竭树脂通过抑制破骨细胞和ROS活性来减轻大鼠SIONFH的发病。这些发现强调了血竭作为早期SIONFH有效药物的潜力,揭示了其对ROS介导的破骨细胞信号通路的治疗影响。