Guizhou University of Traditional Chinese Medicine, Guiyang, 550025, China.
State Key Laboratory of Dao-di Herbs, Beijing, 100700, China.
J Ethnopharmacol. 2025 Jan 30;337(Pt 2):118857. doi: 10.1016/j.jep.2024.118857. Epub 2024 Oct 1.
The Radix Dipsaci, a traditional Chinese medicine with a history spanning over 2000 years in China, is widely recognized for its hepatorenal tonic properties, musculoskeletal fortifying effects, fracture healing capabilities, and its frequent application in the treatment of osteoporosis. Like many traditional Chinese herbal medicines, preparations from Radix Dipsaci are at risk of contamination by harmful mycotoxins such as aflatoxin B1.
This study aims to evaluate the impact of aflatoxin B1 contamination on Radix Dipsaci in terms of changes in quality, efficacy of anti-osteoporosis and hepatorenal toxicity.
The contamination rates and levels of major mycotoxins were determined in 45 batches of Radix Dipsaci samples using UPLC-MS/MS analysis. The total saponin content and the levels of akebia saponin D in Radix Dipsaci and its decoctions were evaluated through high-performance liquid chromatography (HPLC) analysis. Differences in secondary metabolites between samples without any mycotoxin contamination (N-RD) and those contaminated solely by aflatoxin B1 (AFB1-RD) were compared using metabolomics sequencing and analysis. The anti-osteoporotic efficacy of Radix Dipsaci contaminated with aflatoxin B1 was assessed in a murine model of retinoic acid-induced osteoporosis by quantifying bone mineral content and bone mineral density using dual-energy X-ray absorptiometry. Additionally, the hepatorenal toxicity of Radix Dipsaci contaminated with aflatoxin B1 was evaluated using hematoxylin-eosin staining and enzyme-linked immunosorbent assay (ELISA).
The results indicated that aflatoxin B1 (AFB1) was the most frequently detected mycotoxin, found in 37.7% of the Radix Dipsaci samples. AFB1 contamination significantly altered the secondary metabolites of Radix Dipsaci. Specifically, there was a notable decrease in the levels of total saponins and akebia saponin D in the AFB1-contaminated samples, which exhibited a negative correlation with the levels of AFB1 contamination. However, the administration of a water decoction from AFB1-contaminated Radix Dipsaci did not result in significant improvements in bone mineral density, bone mineral salt content, the trabecular number, trabecular area, proportion of trabecular bone volume/tissue volume and trabecular separation in an osteoporosis mouse model. Additionally, we observed that approximately 16.04% of AFB1 could migrate from the raw herbs into the decoction, leading to hepatocyte and kidney cell damage, as well as increased levels of the oxidative stress molecule malondialdehyde and pro-inflammatory cytokines in the liver and kidney tissues of the osteoporosis model mice.
In summary, Radix Dipsaci is highly susceptible to mycotoxin contamination, particularly aflatoxin B1. The contamination of Radix Dipsaci with AFB1 not only impacts their saponin content and anti-osteoporosis effect but also induces hepatotoxicity and nephrotoxicity.
中药续断在中国已有超过 2000 年的历史，被广泛认为具有滋补肝肾、强筋健骨、促进骨折愈合的功效，常用于治疗骨质疏松症。与许多中药一样，续断的制剂也存在被黄曲霉毒素 B1 等有害真菌毒素污染的风险。
本研究旨在评估黄曲霉毒素 B1 污染对续断质量、抗骨质疏松和肝肾毒性的影响。
采用 UPLC-MS/MS 分析检测 45 批续断样品中的主要真菌毒素污染率和污染水平。采用高效液相色谱法（HPLC）分析续断及其煎液中的总皂苷和钩藤皂苷 D 含量。采用代谢组学测序和分析比较无任何真菌毒素污染的样品（N-RD）和仅受黄曲霉毒素 B1 污染的样品（AFB1-RD）之间的次级代谢产物差异。采用双能 X 射线吸收仪（DXA）检测黄曲霉毒素 B1 污染的续断对维甲酸诱导骨质疏松症小鼠模型的抗骨质疏松作用，定量检测骨矿物质含量和骨矿物质密度。此外，采用苏木精-伊红染色和酶联免疫吸附试验（ELISA）检测黄曲霉毒素 B1 污染的续断的肝肾毒性。
结果表明，黄曲霉毒素 B1（AFB1）是最常被检测到的真菌毒素，在 37.7%的续断样品中被发现。AFB1 污染显著改变了续断的次级代谢产物。具体而言，AFB1 污染的样品中总皂苷和钩藤皂苷 D 的含量明显下降，与 AFB1 污染水平呈负相关。然而，在骨质疏松症小鼠模型中，给予黄曲霉毒素 B1 污染的续断水煎液并未显著改善骨矿物质密度、骨矿物质盐含量、骨小梁数量、骨小梁面积、骨小梁体积/组织体积的比例以及骨小梁分离度。此外，我们观察到约 16.04%的 AFB1 可以从生药转移到煎液中，导致肝细胞和肾细胞损伤，以及骨质疏松症模型小鼠肝、肾组织中氧化应激分子丙二醛和促炎细胞因子水平升高。
综上所述，续断极易受到真菌毒素污染，特别是黄曲霉毒素 B1。续断受 AFB1 污染不仅会影响其皂苷含量和抗骨质疏松作用，还会导致肝毒性和肾毒性。
