Glabrol impurity exacerbates glabridin toxicity in zebrafish embryos by increasing myofibril disorganization.
Glabrol impurity exacerbates glabridin toxicity in zebrafish embryos by increasing myofibril disorganization.
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DOI：
10.1016/j.jep.2021.114963
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发表时间：
2021-12
影响因子：
5.4
通讯作者：
Qingquan Guo;Shaojuan Wu;W. Liang;Jianhua Tan;X. Liu;Yuxi Yuan;Xiaohong Li;Haishan Zhao
中科院分区：
文献类型：
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作者：
Qingquan Guo;Shaojuan Wu;W. Liang;Jianhua Tan;X. Liu;Yuxi Yuan;Xiaohong Li;Haishan Zhao
民族药理学相关性光甘草定，从甘草中提取，由于其抗炎和抗氧化活性以及其抑制黑色素合成的能力而被广泛用于治疗色素沉着过度。这导致了对其质量和安全的严格监管。然而，传统的质量控制方法用于植物提取物不能反映产品的质量，由于多个未知杂质，这需要进一步的杂质分析的study. Objective本研究确定了光甘草定的毒性杂质及其毒理机制。使用斑马鱼和细胞模型评价样品毒性。为了鉴定杂质，用超高效液相色谱-四极杆-轨道阱质谱联用技术对不同毒性的样品进行分析。在斑马鱼模型中验证了相关杂质的毒性。鬼笔环肽染色被用来评估肌原纤维align. ResultsGlabridin含量在样品中的细微变化是相似的，有显着差异的毒性。使用四种不同的哺乳动物细胞系验证了结果。在具有最高毒性的样品中鉴定了较高含量的光甘草酮和光甘草醇。在斑马鱼模型中，添加光甘草酚使光甘草定在受精后48、72和96 h的LC 50分别降低至9.224、6.229和5.370 μM，而光甘草酮没有任何毒性作用。鬼笔环肽染色表明，glabrol杂质加剧了glabridin在斑马鱼embryon.ConclusionGlabrol，而不是glabrone的肌肉毒性，被确定为一个关键的杂质，增加glabridin毒性。这一发现表明在光甘草定产品生产过程中控制光甘草酚含量是必要的。
Ethnopharmacological relevanceGlabridin, extracted fromGlycyrrhiza glabraL., is widely used for the treatment of hyperpigmentation because of its anti-inflammatory and antioxidant activities and its ability to inhibit melanin synthesis. This led to the strict regulation of its quality and safety. However, traditional quality control methods used for plant extracts cannot reflect the product quality owing to multiple unknown impurities, which necessitates the further analysis of impurities.Aim of the studyThe study identified the toxic impurities of glabridin and their toxicological mechanism.Materials and methodsIn total, 10 glabridin samples from different sources were quantified using high-performance liquid chromatography. Sample toxicities were evaluated using zebrafish and cell models. To identify impurities, samples with different toxicity were analyzed by ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap mass spectrometry. The toxicity of related impurities was verified in the zebrafish model. Phalloidin stain was used to evaluate subtle changes in myofibril alignment.ResultsAlthough glabridin content in the samples was similar, there were significant differences in toxicity. The results were verified using four different mammalian cell lines. Higher contents of glabrone and glabrol were identified in the sample with the highest toxicity. In the zebrafish model, the addition of glabrol reduced the LC50of glabridin to 9.224, 6.229, and 5.370 μM at 48, 72, and 96 h post-fertilization, respectively, whereas glabrone did not have any toxic effect. Phalloidin staining indicated that a glabrol impurity exacerbates the myotoxicity of glabridin in zebrafish embryos.ConclusionGlabrol, but not glabrone, was identified as a key impurity that increased glabridin toxicity. This finding indicates that controlling glabrol content is necessary during glabridin product production.