A spiny plant long used in folk medicine has now shown a measurable ability to blunt genetic damage in laboratory tests. Researchers report that crude extract of Echinops spinosus, the spiny globe thistle, and its aqueous fraction reduced chromosomal aberrations and DNA strand breaks triggered by the mutagen ethyl methanesulfonate in mouse bone marrow and spleen cells. The findings, published in the journal Genes and Environment, mark the first examination of the genoprotective effects of this plant using standard genotoxicity assays in mouse tissues.
Echinops spinosus belongs to the Asteraceae family, one of the largest flowering plant families with roughly 1600 genera and 25,000 species. The Echinops genus alone contains more than 130 species spread across Eurasia, North Africa and East Africa. Traditional medicine has relied on the plant for its anti-inflammatory, diuretic and digestive properties, and it has been used to treat ailments including splenic and renal disorders. Until now, however, no study had tested whether the plant could protect genetic material from a known mutagen.
The research team, led by scientists including Kawthar A. Diab and Zeinab M. Hassan, chose ethyl methanesulfonate as the damaging agent. EMS is a monofunctional alkylating agent used extensively in DNA repair studies as a mutagenic, teratogenic and carcinogenic compound. Its ethyl group reacts with nucleophilic sites on DNA, producing ethylated nucleotides at nitrogen atoms such as N7-guanine and N3-adenine, and to a lesser extent at oxygen atoms such as O6-guanine. Alkylated nitrogen atoms cause chromosomal aberrations, while alkylated oxygen atoms drive substitution point mutations when O6-ethylguanine mispairs with thymine during replication.
To prepare the test materials, the investigators powdered 700 grams of the aerial flowering parts of the plant and extracted them with 70 percent ethanol, yielding 64.17 grams of crude extract, designated CEES. This crude extract was then suspended in hot water and partitioned successively with methylene chloride, n-butanol and distilled water. The final aqueous residue produced 41.82 grams of the aqueous fraction, designated AFES. The team then characterized both preparations chemically using gas chromatography-mass spectrometry, high performance liquid chromatography and colorimetric assays for total polyphenol and flavonoid contents.
The chemical profiling revealed distinct differences between the two preparations. GC-MS analysis identified 25 primary metabolites in the aqueous fraction, with nitrogenous compounds and amino acids dominating at 77.13 percent of total silylated metabolites and the compound bis(trimethylsilyl) ethylamine serving as the primary constituent at 50.05 percent. HPLC analysis detected 17 secondary compounds in the crude extract and 14 in the aqueous fraction. Chlorogenic acid was the main constituent in both samples, accounting for 42.3 percent of total compounds in the crude extract and 58.8 percent in the aqueous fraction. The crude extract also contained rosmarinic acid, ferulic acid and gallic acid as major components. Colorimetric measurements showed the crude extract carried a 3.5-fold higher total polyphenol content and a 3.1-fold higher total flavonoid content than the aqueous fraction.
Before testing protection, the team confirmed the extracts themselves were not harmful. In cytotoxicity assays using a colorimetric cell count kit, neither CEES nor AFES reduced viability in primary mouse bone marrow and spleen cultures after 24 hours of exposure at concentrations up to 1000 micrograms per milliliter. In fact, both treatments, particularly at the highest concentration, increased cell viability, suggesting proliferative or mitogenic activity from the phytochemicals present. In vivo, a single oral dose of the aqueous fraction at 5000 milligrams per kilogram produced no signs of death or toxicity in male mice during an initial 24-hour observation or a 14-day follow-up, and the crude extract had previously shown an oral LD50 greater than 5000 milligrams per kilogram.
The in vitro experiments used cultured primary mouse bone marrow and spleen cells treated with two concentrations of either extract, 250 or 500 micrograms per milliliter, for 24 hours, followed by EMS at 300 micrograms per milliliter for two hours before harvest. EMS alone markedly increased chromosomal aberrations in bone marrow cultures, reaching 19.33 percent, a 7.2-fold rise over controls, and in splenic cultures, reaching 16.33 percent, a 4.9-fold rise. Breaks and fragments were the predominant aberration types. Pretreatment with the higher concentration of either extract significantly attenuated this chromosomal damage, achieving reduction indices of 34.00 percent in bone marrow and 46.15 percent in spleen cultures, while the lower concentration produced only marginal reductions.
The comet assay, which quantifies DNA fragmentation, told a similar story. EMS treatment raised the olive tail moment and percentage tail DNA in bone marrow cultures by 10.93 percent, a 2.9-fold increase, and in spleen cultures by 7.49 percent, a 1.72-fold increase. A 24-hour pretreatment with either extract significantly reduced comet tail formation. The crude extract at 500 micrograms per milliliter was more effective, restoring percentage tail DNA in bone marrow to 4.71 percent, a 2.3-fold decrease, and in spleen to 4.57 percent, a 1.6-fold decrease, approaching control levels.
The in vivo experiments strengthened the protective signal. Forty adult male Swiss albino mice were divided into eight groups of five. Some groups received the extracts orally at 250 or 500 milligrams per kilogram for seven consecutive days, with or without a single intraperitoneal injection of EMS at 300 milligrams per kilogram. EMS alone increased chromosomal aberrations to 27.0 percent, a 7.5-fold rise, and micronucleated polychromatic erythrocytes to 14.14 percent, a 16.8-fold rise, in bone marrow cells. Breaks, fragments and Robertsonian translocations predominated. When mice received the extracts before EMS, these genetic damages fell considerably. The high dose of crude extract produced reductions of 54 percent in chromosomal aberrations and 56 percent in micronuclei, while the high dose of the aqueous fraction achieved reductions of 50 percent and 49 percent respectively. Individual treatment with either extract alone did not affect chromosomal aberration or micronucleus levels compared to controls.
The authors conclude that highly polar constituents primarily mediated the antimutagenic activity, while other phytoconstituents in the crude extract, including moderately polar and nonpolar compounds, synergistically potentiated the genoprotective effect, explaining why the crude extract outperformed the aqueous fraction. They also note the protection was more pronounced in vivo than in vitro, possibly because metabolic activation of the phytochemicals and prolonged exposure allowed cumulative protective effects. The study has limitations the researchers acknowledge: they did not compare other organic fractions of the plant, which prevents identification of the most effective fraction and isolation of the primary protective compounds, and the underlying protective mechanisms, such as antioxidant effects or activation of DNA repair pathways, remain unclear and require further study. The work was conducted following National Institute of Health guidelines and approved by the Institutional Ethics Committee of the National Research Centre in Cairo, Egypt.
Subject of Research: Antimutagenic activity of Echinops spinosus extracts against EMS-induced genetic damage in mice
Article Title: In vitro and in vivo antimutagenic activity of Echinops spinosus crude extract and its aqueous fraction in mouse bone marrow and spleen
Article References: Diab, K. A., Farghaly, A. A., Hassan, E. E., Fahmy, M. A., Hassan, E. M., & Hassan, Z. M. (2025). In vitro and in vivo antimutagenic activity of Echinops spinosus crude extract and its aqueous fraction in mouse bone marrow and spleen. Genes and Environment, 47(1), Article 21. https://doi.org/10.1186/s41021-025-00341-z
Image Credits: AI Generated
DOI: 10.1186/s41021-025-00341-z
Keywords: Echinops spinosus, antimutagenic, ethyl methanesulfonate, chromosomal aberrations, comet assay, micronucleus, mouse bone marrow, spleen, chlorogenic acid, phytochemicals, genoprotection, traditional medicine
News Source: Drew Townsend. (November 5, 2026). Spiny Globe Thistle Extract Shields Mouse Bone Marrow Cells from Mutagen Damage. Scienmag.
