Department of Orthopedics, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Hualien, 97002, Taiwan; Institute of Medical Science, Tzu Chi University, Hualien, 97004, Taiwan; School of Medicine, Tzu Chi University, Hualien, 97004, Taiwan.
Department of Orthopedics, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Hualien, 97002, Taiwan.
J Ethnopharmacol. 2022 Oct 28;297:115565. doi: 10.1016/j.jep.2022.115565. Epub 2022 Jul 19.
Gu Sui Bu (GSB), the dried rhizome of Drynaria fortunei J. Sm., is widely used in traditional Chinese medicine for treating fractures and osteoporosis. Although glucocorticoids are widely prescribed in modern medicine, the efficacy of GSB in treating glucocorticoid-induced osteoporosis (GIOP) remains unclear.
GIOP is one of the most prevalent forms of osteoporosis and increases the risk of fracture, which can cause severe complications in elderly people. Safe, efficacious, and cost-effective treatment options for GIOP are thus warranted. The present study investigated the efficacy and mechanism of GSB for treating GIOP.
We established an efficient and robust in vivo GIOP model by optimizing zebrafish larvae rearing conditions and the dose and duration of dexamethasone treatment. Bone calcification was evaluated through calcein staining. To quantify the degree of vertebral mineralization in the larvae, we developed a scoring system based on the rate of vertebral calcification; this system reduced quantification errors among individual zebrafish caused by inconsistencies in staining or imaging parameters. Quantitative real-time polymerase chain reaction was used to access the expression levels of genes essential to the differentiation and function of bone cells. High-performance liquid chromatography was employed to identify naringin in the GSB extract.
GSB significantly reversed the dexamethasone-induced calcification delay in zebrafish larvae. GSB enhanced osteoblast activity by increasing the expression of collagen I, osteopontin, and osteonectin and repressed bone resorption by decreasing the expression of matrix metalloproteinases (mmps), including mmp9 and mmp13a. We also identified naringin as one of the constituents of GSB responsible for the herbal extract's anti-GIOP activity.
Using the in vivo zebrafish GIOP model that we established, the efficacy of traditional Chinese medicines in treating GIOP could be systematically investigated. GSB has an osteogenic effect and may thus be an efficacious and cost-effective treatment option for GIOP. Notably, bone resorption activity was found to be retained after GSB treatment, which would be beneficial for maintaining normal bone remodeling.
骨碎补(GSB),是水龙骨科植物槲蕨的干燥根茎,在中医中广泛用于治疗骨折和骨质疏松症。虽然糖皮质激素在现代医学中广泛应用,但 GSB 治疗糖皮质激素诱导性骨质疏松症(GIOP)的疗效尚不清楚。
GIOP 是最常见的骨质疏松症形式之一,增加了骨折的风险,这会给老年人造成严重的并发症。因此,需要安全、有效且具有成本效益的 GIOP 治疗选择。本研究旨在探讨 GSB 治疗 GIOP 的疗效和机制。
我们通过优化斑马鱼幼虫饲养条件以及地塞米松的剂量和作用时间,建立了一种高效、稳健的体内 GIOP 模型。通过 calcein 染色评估骨钙化。为了量化幼虫脊柱矿化程度,我们开发了一种基于脊柱钙化率的评分系统;该系统减少了因染色或成像参数不一致而导致个体斑马鱼之间定量误差。采用实时定量聚合酶链反应(qPCR)检测对骨细胞分化和功能至关重要的基因表达水平。采用高效液相色谱法(HPLC)鉴定 GSB 提取物中的柚皮苷。
GSB 显著逆转了地塞米松诱导的斑马鱼幼虫钙化延迟。GSB 通过增加胶原蛋白 I、骨桥蛋白和骨连接蛋白的表达来增强成骨细胞活性,并通过降低基质金属蛋白酶(MMPs)的表达来抑制骨吸收,包括 MMP9 和 MMP13A。我们还鉴定出柚皮苷是 GSB 中一种负责其抗 GIOP 活性的成分。
使用我们建立的体内斑马鱼 GIOP 模型,可以系统地研究传统中药治疗 GIOP 的疗效。GSB 具有成骨作用,因此可能是治疗 GIOP 的一种有效且具有成本效益的选择。值得注意的是,GSB 治疗后发现骨吸收活性得到保留,这将有利于维持正常的骨重塑。