Black cohosh extract (BCE) is a popular botanical dietary supplement marketed to relieve symptoms of various gynecological ailments. Studies conducted by the National Toxicology Program (NTP) showed that BCE induces micronucleated erythrocytes in female rats and mice. Subsequently, the NTP showed that a variety of BCEs, including the sample that induced micronuclei (MN) in vivo ("NTP BCE") had a similar effect in human TK6 cells. Further testing with the MultiFlow® DNA Damage Assay revealed that TK6 cells exposed to NTP BCE, as well as a BCE reference material (BC XRM), exhibited a signature consistent with aneugenic activity in TK6 cells. Results from experiments reported herein confirmed these in vitro observations with NTP BCE and BC XRM. We extended these studies to include a novel test system, the MultiFlow Aneugen Molecular Mechanism Assay. For these experiments, TK6 cells were exposed to NTP BCE and BC XRM over a range of concentrations in the presence of fluorescent Taxol (488 Taxol). After 4 h, nuclei from lysed cells were stained with a nucleic acid dye and labeled with fluorescent antibodies against phospho-histone H3 (p-H3) and Ki-67. Whereas BCEs did not affect p-H3:Ki-67 ratios (a signature of aneugenic mitotic kinase inhibitors), 488 Taxol-associated fluorescence (a tubulin binder-sensitive endpoint) was affected. More specifically, 488 Taxol-associated fluorescence was reduced over the same concentration range that was previously observed to induce MN. These results provide direct evidence that BCEs destabilize microtubules in vitro, and this is the molecular mechanism responsible for the aneugenicity findings. Environ. Mol. Mutagen. 2019. © 2019 The Authors. Environmental and Molecular Mutagenesis published by Wiley Periodicals, Inc. on behalf of Environmental Mutagen Society.

中文翻译：

---

黑升麻提取物诱导微核的气生作用机制的证据。

黑升麻提取物（BCE）是一种流行的植物性膳食补充剂，旨在缓解各种妇科疾病的症状。美国国家毒理学计划（NTP）进行的研究表明，BCE可以诱导雌性大鼠和小鼠的微核红细胞。随后，NTP显示各种BCE，包括在体内诱导微核（MN）的样品（“ NTP BCE”）在人TK6细胞中具有相似的作用。DNA损伤测定法的进一步测试表明，暴露于NTP BCE的TK6细胞以及BCE参考材料（BC XRM）均显示出与TK6细胞中的造血活性一致的特征。本文报道的实验结果证实了使用NTP BCE和BC XRM进行的这些体外观察。我们将这些研究扩展到包括新颖的测试系统，MultiFlow中子分子机理分析。对于这些实验，在存在荧光紫杉醇（488紫杉醇）的情况下，将TK6细胞以一定浓度范围暴露于NTP BCE和BC XRM。4小时后，将裂解细胞的细胞核用核酸染料染色，并用针对磷酸组蛋白H3（p-H3）和Ki-67的荧光抗体标记。尽管BCE不会影响p-H3：Ki-67的比率（非致病性有丝分裂激酶抑制剂的标志），但488个紫杉醇相关的荧光（微管蛋白结合物敏感的终点）受到影响。更具体地，在先前观察到诱导MN的相同浓度范围内，减少了488紫杉醇相关的荧光。这些结果提供了直接的证据，证明BCE在体外会破坏微管的稳定性，而这是导致无气性发现的分子机制。环境。大声笑 诱变剂。2019.©2019 The Authors。Wiley Periodicals，Inc.代表环境诱变学会出版的《环境和分子诱变》。