Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran.
J Ethnopharmacol. 2021 Feb 10;266:113428. doi: 10.1016/j.jep.2020.113428. Epub 2020 Oct 1.
The application of the herb Ziziphora clinopodioides Lam. in folk medicine and as a food additive has been recommended due to its many claimed bioactivities. Regardless of the plant benefits, its safety considerations are largely unknown.
The aim of the present research was to determine the chemical compositions and cytotoxicity, genotoxicity, and mutagenicity potentials of the ethanolic extract of Ziziphora clinopdioides Lam. (EEZC).
GC-MS and LC-MS analysis were used for chemical composition determination. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and trypan blue exclusion dye assays were used for cytotoxicity and the Comet assay was employed for genotoxicity assessment on human blood lymphocytes. Also, the Ames Salmonella/microsome test was carried out for the evaluation of mutagenicity.
Pulegone was the main component of the n-hexane fraction. Different phenolic acids and flavonoids were detected by LC-MS. The cytotoxicity study indicated a conspicuous decline in human lymphocyte viability ranging from 52% to 100% as showed by the MTT assay and 67% up to 100% by the trypan blue assay, at 1 and 10 mg/mL, respectively. The Comet assay results revealed a dose dependent genotoxicity, in so much as 90% and 98% of the cells were screened as damaged at concentrations of 5 and 10 mg/mL, respectively. An incidence rate of 8% and 13% of grade 4 damage was observed at 5 and 10 mg/mL, respectively. Additionally, the DNA damage index (DI) was elevated dose-dependently by a rising concentration of the extract, wherein the DI at 10 mg/mL concentration was 2.22, which was 22 times greater than that of negative control, and even more than positive control. The Ames test exhibited no signs of mutagenicity for neither Salmonella typhimurium TA98 nor TA100 strains, accompanied or unaccompanied by S9 metabolic activation.
Results indicated a dose-dependent cytotoxicity and genotoxicity potential of the EEZC on human lymphocytes, suggesting that this plant should be used with caution by consumers, even in the food and pharmaceutical industries. Since the plant usage in daily life continues to increase due to its ever growing phytotherapical and phytonutritional properties, it may pose a health risk by its high concentration's uptake. Although no mutagenicity of this extract was observed in this study, further research is recommended to clarify the mutagenic risks of this herb.
由于其许多声称的生物活性，唇形科植物荆芥（Ziziphora clinopodioides Lam.）在民间医学和作为食品添加剂中的应用已得到推荐。无论该植物有何益处，其安全性考虑因素在很大程度上仍是未知的。
本研究旨在确定荆芥（Ziziphora clinopdioides Lam.）乙醇提取物（EEZC）的化学成分以及细胞毒性、遗传毒性和致突变潜力。
采用 GC-MS 和 LC-MS 分析进行化学成分测定。MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐）和台盼蓝排斥染料测定法用于细胞毒性评估，彗星试验用于评估人血淋巴细胞的遗传毒性。此外，还进行了 Ames 沙门氏菌/微粒体试验以评估致突变性。
正己烷部分的主要成分是薄荷酮。通过 LC-MS 检测到不同的酚酸和类黄酮。MTT 测定和台盼蓝测定分别显示，1 和 10mg/mL 时，人淋巴细胞活力明显下降，范围为 52%至 100%；分别为 67%至 100%。彗星试验结果显示，随着浓度的增加，遗传毒性呈剂量依赖性，分别在 5 和 10mg/mL 时，90%和 98%的细胞被筛选为受损。在 5 和 10mg/mL 时，分别观察到 8%和 13%的 4 级损伤发生率。此外，提取物浓度升高时，DNA 损伤指数（DI）呈剂量依赖性升高，其中 10mg/mL 浓度下的 DI 为 2.22，是阴性对照的 22 倍，甚至高于阳性对照。Ames 试验表明，无论是添加或不添加 S9 代谢活化，荆芥乙醇提取物对鼠伤寒沙门氏菌 TA98 和 TA100 菌株均无致突变性。
结果表明，荆芥乙醇提取物对人淋巴细胞具有剂量依赖性的细胞毒性和遗传毒性潜力，这表明即使在食品和制药行业，消费者也应谨慎使用该植物。由于其日益增长的植物疗法和植物营养素特性，这种植物在日常生活中的使用不断增加，因此其高浓度摄入可能会带来健康风险。尽管本研究未观察到该提取物的致突变性，但建议进一步研究以阐明该草药的致突变风险。
