Efficacy of the herbal pair, Radix Achyranthis Bidentatae and Eucommiae Cortex, in preventing glucocorticoid-induced osteoporosis in the zebrafish model
Efficacy of the herbal pair, Radix Achyranthis Bidentatae and Eucommiae Cortex, in preventing glucocorticoid-induced osteoporosis in the zebrafish model
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牛膝和杜仲这对草药对预防斑马鱼模型中糖皮质激素诱导的骨质疏松症的功效
DOI：
10.1016/j.joim.2021.11.003
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发表时间：
2022-01-04
影响因子：
4.8
通讯作者：
Dai, Wei-Wei
中科院分区：
文献类型：
--
作者：
Lee, Joon-Ho;Wei, Yuan-Ji;Dai, Wei-Wei
目的：在传统中医中，牛膝（RAB）和杜仲（EC）这对草药被广泛用于治疗骨质疏松症。在此，我们确定这对草药是否可用于改善糖皮质激素（GC）诱导的骨质疏松症（GIOP），并找到其在斑马鱼中的最佳剂量。方法：使用高效液相色谱分别表征 RAB 和 EC 水提取物的特性。将受精后 5 天的斑马鱼幼虫暴露于 10 μmol/L 地塞米松 (Dex) 96 小时，诱导其骨质疏松。共同施用七种不同比例的 RAB 和 EC 组合。通过计算斑马鱼椎体面积和总亮度、茜素红染色以及检测碱性磷酸酶 (ALP) 活性来确定治疗效果。通过多元回归分析来测试最佳剂量比例。结果：根据中国药典（2015年版），β-蜕皮激素（β-Ecd）是RAB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EC提取物中的主要生物活性标志物。 β-Ecd和PDG含量值均符合中国药典标准。 10 μmol/L Dex 处理减少了斑马鱼的椎面积、总亮度和 ALP 活性，但 RAB 和 EC 减弱了这些影响。 50 μg/mL RAB 和 50 μg/mL EC 组合是预防斑马鱼 GIOP 的最佳方法。采用逆转录定量聚合酶链反应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)、成骨蛋白-1 (OP-1)、骨含γ-羧基谷氨酸蛋白 (BGLAP) 和 β-连环蛋白水平。 RAB 和 EC 联合治疗抵消了这种效应（P < 0.05）。此外，两种草药提取物一起使用的效果比单独使用单一草药治疗的效果更好。这些结果表明，在 GC 存在的情况下，RAB 和 EC 可以保留成骨细胞功能。最佳质量比为1：1。结论：RAB与EC中药组合可改善斑马鱼GC诱导的效应，最佳剂量比为1：1。 (C)2022 上海中医药大学附属上海岳阳医院。由 Elsevier B.V. 出版。保留所有权利。
Objective: 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.Methods: 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 mu mol/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.Results: According to the Chinese Pharmacopoeia (2015), beta-ecdysone (beta-Ecd) is a major bioactive marker in RAB extract, while pinoresinol diglucoside (PDG) is the major marker in EC extract. Both of beta-Ecd and PDG content values aligned with the Chinese Pharmacopoeia standards. Treatment with 10 mu mol/L Dex reduced zebrafish vertebral area, sum brightness, and ALP activity, but RAB and EC attenuated these effects. Combining 50 mu g/mL RAB and 50 mu g/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 mu mol/L Dex decreased runt-related transcription factor 2 (Runx2), osteogenic protein-1 (OP-1), bone gamma-carboxyglutamic acid-containing protein (BGLAP), and beta-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.Conclusion: RAB and EC herbal pair could ameliorate GC-induced effects in zebrafish, with 1:1 as the optimal dosage ratio. (C) 2022 Shanghai Yueyang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine. Published by Elsevier B.V. All rights reserved.