Central Laboratory of Science and Technology Department, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.
Cardiovascular Department, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.
J Integr Med. 2022 Jan;20(1):83-90. doi: 10.1016/j.joim.2021.11.003. Epub 2021 Nov 7.
In traditional Chinese medicine, the herbal pair, Radix Achyranthis Bidentatae (RAB) and Eucommiae Cortex (EC), is widely used to treat osteoporosis. Herein, we determined whether this herbal pair can be used to ameliorate glucocorticoid (GC)-induced osteoporosis (GIOP) and find its optimal dosage in zebrafish.
The characteristics of the aqueous extract of RAB and EC were separately characterized using high-performance liquid chromatography. Osteoporosis was induced in 5-day post-fertilization zebrafish larvae by exposing them to 10 μmol/L dexamethasone (Dex) for 96 h. Seven combinations of different ratios of RAB and EC were co-administered. Treatment efficacy was determined by calculating zebrafish vertebral area and sum brightness, via alizarin red staining, and by detecting alkaline phosphatase (ALP) activity. Multiple regression analysis was conducted to test the optimal dosage ratio.
According to the Chinese Pharmacopoeia (2015), β-ecdysone (β-Ecd) is a major bioactive marker in RAB extract, while pinoresinol diglucoside (PDG) is the major marker in EC extract. Both of β-Ecd and PDG content values aligned with the Chinese Pharmacopoeia standards. Treatment with 10 μmol/L Dex reduced zebrafish vertebral area, sum brightness, and ALP activity, but RAB and EC attenuated these effects. Combining 50 µg/mL RAB and 50 µg/mL EC was optimal for preventing GIOP in zebrafish. Reverse transcription-quantitative polymerase chain reaction was used to evaluate the mRNA expression of osteogenesis-related genes. A treatment of 10 μmol/L Dex decreased runt-related transcription factor 2 (Runx2), osteogenic protein-1 (OP-1), bone γ-carboxyglutamic acid-containing protein (BGLAP), and β-catenin levels. This effect was counteracted by RAB and EC co-treatment (P < 0.05). Additionally, the effect of using the two herbal extracts together was better than single-herb treatments separately. These results demonstrated that RAB and EC preserve osteoblast function in the presence of GC. The best mass ratio was 1:1.
RAB and EC herbal pair could ameliorate GC-induced effects in zebrafish, with 1:1 as the optimal dosage ratio.
在中医中，牛膝和杜仲这对草药被广泛用于治疗骨质疏松症。本研究旨在确定该草药组合是否可用于改善糖皮质激素（GC）诱导的骨质疏松症（GIOP），并在斑马鱼中找到其最佳剂量。
分别采用高效液相色谱法对牛膝和杜仲水提物的特征进行了表征。通过将 5 日龄斑马鱼胚胎暴露于 10 μmol/L 地塞米松（Dex）中 96 h 来诱导骨质疏松症。共给予 7 种不同比例的牛膝和杜仲组合。通过茜素红染色计算斑马鱼椎骨面积和总和亮度，并通过检测碱性磷酸酶（ALP）活性来确定治疗效果。采用多元回归分析来检验最佳剂量比。
根据《中国药典》（2015 年版），β-蜕皮甾酮（β-Ecd）是牛膝提取物中的主要生物活性标志物，而松脂醇二葡萄糖苷（PDG）是杜仲提取物中的主要标志物。β-Ecd 和 PDG 的含量均符合《中国药典》标准。10 μmol/L Dex 处理降低了斑马鱼的椎骨面积、总和亮度和 ALP 活性，但牛膝和杜仲可减轻这些影响。以 50 μg/mL 牛膝和 50 μg/mL 杜仲联合治疗对预防斑马鱼的 GIOP 最为有效。采用逆转录定量聚合酶链反应（qRT-PCR）评估成骨相关基因的 mRNA 表达。10 μmol/L Dex 处理降低了 runt 相关转录因子 2（Runx2）、骨形成蛋白 1（OP-1）、骨γ-羧基谷氨酸蛋白（BGLAP）和β-连环蛋白的水平。牛膝和杜仲联合治疗可逆转这种作用（P<0.05）。此外，两种草药提取物联合使用的效果优于单独使用一种草药。这些结果表明，牛膝和杜仲在 GC 存在的情况下可维持成骨细胞功能。最佳质量比为 1:1。
牛膝和杜仲草药组合可改善 GC 诱导的斑马鱼作用，最佳剂量比为 1:1。
Zotero 插件