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[This corrects the article DOI: 10.3389/fphar.2024.1394557.].

Alchemilla vulgaris L. (lady’s mantle) was used for centuries in Europe and Balkan countries for treatments of numerous conditions and diseases of the reproductive system, yet some of the biological activities of lady’s mantle have been poorly studied and neglected. The present study aimed to estimate the potential of A. vulgaris ethanolic extract from Southeast Serbia to prevent and suppress tumor development in vitro, validated by antioxidant, genoprotective, and cytotoxic properties. A total of 45 compounds were detected by UHPLC–HRMS analysis in A. vulgaris ethanolic extract. Measurement of antioxidant activity revealed the significant potential of the tested extract to scavenge free radicals. In addition, the analysis of micronuclei showed an in vitro protective effect on chromosome aberrations in peripheral human lymphocytes. A. vulgaris extract strongly suppressed the growth of human cell lines derived from different types of tumors (MCF-7, A375, A549, and HCT116). The observed antitumor effect is realized through the blockade of cell division, caspase-dependent apoptosis, and autophagic cell death. Our study has shown that Alchemilla vulgaris L. is a valuable source of bioactive compounds able to protect the subcellular structure from damage, thus preventing tumorigenesis as well as suppressing tumor cell growth.

Introduction: Isoproterenol (ISO) is regarded as an adrenergic non-selective β agonist. It regulates myocardial contractility and may cause damage to cardiac tissues. Alchemilla vulgaris (AV) is an herbal plant that has garnered considerable attention due to its anti-inflammatory and antioxidant bioactive components. The present investigation assessed the cardioprotective potential of AV towards ISO-induced myocardial damage. Methods: Four groups of mice were utilized: control that received saline, an ISO group (85 mg/kg, S.C.), ISO + AV100, and ISO + AV200 groups (mice received 100 or 200 mg/kg AV orally along with ISO). Results and discussion: ISO induced notable cardiac damage demonstrated by clear histopathological disruption and alterations in biochemical parameters. Intriguingly, AV treatment mitigates ISO provoked oxidative stress elucidated by a substantial enhancement in superoxide dismutase (SOD) and catalase (CAT) activities and reduced glutathione (GSH) content, as well as a considerable reduction in malondialdehyde (MDA) concentrations. In addition, notable downregulation of inflammatory biomarkers (IL-1β, TNF-α, and RAGE) and the NF-κB/p65 pathway was observed in ISO-exposed animals following AV treatment. Furthermore, the pro-apoptotic marker Bax was downregulated together with autophagy markers Beclin1 and LC3 with in ISO-exposed animals when treated with AV. Pre-treatment with AV significantly alleviated ISO-induced cardiac damage in a dose related manner, possibly due to their antioxidant and anti-inflammatory properties. Interestingly, when AV was given at higher doses, a remarkable restoration of ISO-induced cardiac injury was revealed.

Methyl methanesulfonate (MMS) is a chemical compound that exerts an alkylating effect on DNA and proteins. Alchemilla vulgaris L. is a medicinal plant known for its rich bioactive composition and pharmacological potential. This study investigated the cytoprotective activity of A. vulgaris extract (AVE) against MMS-induced toxicity in Allium cepa L. bulbs. For this purpose, A. cepa bulbs were divided into groups ( n  = 50 per group) and treated with tap water (control), AVE (250 mg/L and 500 mg/L), MMS (4000 µM), or combinations of MMS (4000 µM) with AVE (250 mg/L and 500 mg/L) for 72 h. A comprehensive analysis was conducted to ascertain the impact of the solutions on physiological parameters (rooting percentage, root elongation, and weight gain), cytogenetic endpoints (mitotic index, micronucleus formation, and chromosomal aberrations), biochemical markers (malondialdehyde, superoxide dismutase, catalase, chlorophyll a and b), and meristematic tissue integrity. Molecular docking was employed to elucidate the potential interactions between MMS and key cellular targets, including and tubulins, topoisomerases, glutamate-1-semialdehyde aminotransferase and protochlorophyllide reductase. A targeted LC–MS/MS analysis was carried out to determine the main phenolic compounds that may underlie the observed biological effects. The analysis showed that the extract predominantly contained rosmarinic acid, catechin, p-coumaric acid, rutin, and gentisic acid. MMS administration resulted in a decline in rooting percentage, weight gain, root elongation, mitotic index and chlorophyll content. MMS exposure increased frequencies of micronuclei, chromosomal aberrations, malondialdehyde levels, and the activities of superoxide dismutase and catalase. Meristematic tissue damage in the MMS group included epidermal cell deformation, flattened nuclei, cortical damage, and thickened cortex cell walls. Co-application of AVE significantly alleviated MMS-induced toxicity in a dose-dependent manner. These findings suggest that AVE exhibits notable antigenotoxic and antioxidant properties. However, further in vivo studies are needed to support its potential use in therapeutic or dietary contexts.

Cataract represents one of the leading causes of blindness worldwide and is primarily attributed to protein glycation, oxidative stress, and aggregation of lens crystallins. Surgical lens extraction remains the standard treatment, which, despite its effectiveness, carries potential postoperative risks and economic burdens, thereby underscoring the need for alternative, non-surgical therapeutic approaches. This study aimed to develop a green-synthesized nanosystem based on gold nanoparticles (AuNPs) using Alchemilla vulgaris L. (Lady’s mantle) extract, aiming to achieve efficient antioxidant delivery and sustained therapeutic retention within ocular tissues. Gold nanoparticles were synthesized via the reduction of chloroauric acid (HAuCl·3H 2 O) using the plant extract under ultrasonic agitation and dispersion, yielding AuNPs formulations 1%, 4%, and 7% (w/v). The synthesized nanocomposites were characterized using a UV–Vis spectroscopy, FTIR, XRD, FE-SEM, AFM, and zeta potential analysis. Bio activity was performed using DPPH radical scavenging, MTT assays for cytotoxicity testing, in vitro drug-release studies, and ex-vivo lens transparency assessments, all supported by computational molecular docking and ADMET (absorption, distribution, metabolism, and excretion) analyses. The results demonstrated that NAC-loaded gold nanoparticles (NAC–AuNPs) exhibited enhanced antioxidant activity, achieving 72–89% DPPH inhibition, and a high encapsulation efficiency of 86.1%. The drug-release profile exhibited a higher release rate observed under mildly acidic conditions (pH 6). Ex-vivo experiments using human cataractous lenses revealed a dose-dependent improvement in lens optical clarity at NAC concentrations of 0.05, 0.1, and 0.3 mM, loaded onto 7% (w/v) AuNPs formulations. Molecular docking studies suggested potential hydrogen-bond interactions between NAC and key lens crystallin proteins, providing mechanistic insight into possible structural stabilization of lens crystallin proteins. The green-synthesized AuNP–NAC nanosystem may represents a promising and biocompatible nanocarrier strategy that warrants further in vivo validation.

Living mulches, particularly when designed with officinal plants, beside competing against weeds of a fruit crop, could provide other ecosystem services. The impact on soil chemical and biological features of Alchemilla vulgaris , Fragaria vesca and Mentha x piperita after four years of establishment as understorey living mulches in an organic apple orchard was evaluated. Soil nutrients level was not affected by the living mulches. The combined effect of season and living mulch affected bacterial activity, which reverberated on phyla and taxa abundance. Only few genera were unique to each living mulch species, but the abundance of 213 genera was differentially modified by the living mulches. The soil level of P and K, the metabolism of several C sources, and the bacterial activity and Shannon Index were highly correlated with the relative abundance of about a third of identified bacterial taxa. The analysis of bacterial abundance data gathered since the establishment of the trial was showing a different trend in shaping bacterial soil microbiome depending on the living mulch species.

Ethnomedicinal records have long mentioned the historical usage of Alchemilla vulgaris L. in folk medicine, particularly for the treatment of gynecological issues. Building on this ethnomedicinal knowledge regarding female illnesses, the aim of this research was to evaluate the impact of ethanolic extract of A. vulgaris on mouse breast cancer cells (4T1) in vitro and in vivo, in addition to its effect on the immune compartment in the tumor microenvironment. Behind viability decrease of 4T1 cells induced by treatment with A. vulgaris extract was strong inhibition of cell proliferation accompanied by caspase-dependent apoptosis and autophagic cell death. Observed changes in 4T1 cell culture after treatment were well orchestrated and led to a reduction in metastatic potential through weakened adhesion, invasion, migration, and colony-forming abilities in vitro. Enhanced intracellular production of reactive oxygen and nitrogen species promoted by the treatment might interfere with all the observed effects. Apart from the direct effect on tumor cells, the A. vulgaris extract significantly reduced tumor growth in the solid orthotropic mammary carcinoma model through restitution of efficient local and systemic immune response reflected in enhanced antigen-presenting potential of dendritic cells (DCs) as well as the extent and activity of effector T cells.

Background: Zebrafish (Danio rerio) embryos are transparent in early stages of embryonic development; however, pigment formation at later stages hinders internal organ visualization during imaging. Chemicals such as 1-phenyl-2-thiourea (PTU) and kojic acid, used to block pigmentation, pose significant toxicity risks to human health. Therefore, effective and risk-free depigmentation agents are needed. This study investigates the efficacy of Alchemilla vulgaris (Lady’s mantle) as a safe, natural alternative for zebrafish embryo depigmentation. Methods: A. vulgaris was extracted using four solvents of varying polarities and evaluated for depigmentation efficacy and toxicity. Gas chromatography–mass spectrometry (GC–MS) was used to identify major constituents of the extract. Results: Ethyl acetate extract was more effective at removing pigments than all other extracts, and exhibited the lowest toxicity compared to PTU and kojic acid. Ethyl acetate extract of A. vulgaris remained effective even when administered 48 h post fertilization (post-pigmentation), making it suitable for long-term experiments requiring optical clarity. GC-MS revealed that this extract was rich in linoleic acid, various fatty acid esters, and phenolics, which likely contributed to its depigmentation activity. Conclusions: Based on these findings, we propose that ethyl acetate extract of A. vulgaris is a safer, natural alternative to PTU and kojic acid for depigmenting zebrafish embryos, particularly in long-term imaging experiments. The extract exhibits high efficacy at low concentrations, accompanied by a favorable toxicity profile, demonstrating potential as a depigmentation agent during early zebrafish development. However, further studies are needed to elucidate its mechanism of action.

Research background. The current changes in the global economy, characterised by the climate crisis and the economic and health impact of the COVID-19 pandemic, have led to a significant demand for medicinal herbs. This trend is expected to increase significantly by 2050. In this study, we investigated the biopotential of aqueous infusions of four medicinal plants: Calendula officinalis, Chelidonium majus, Teucrium chamaedrys and Alchemilla vulgaris. Experimental approach. The flavonoid analysis of the aqueous infusions of the selected plants was carried out using the RP-HPLC technique. The antiproliferative activity of the prepared aqueous plant infusions was analysed against three human cancer cell lines (MDA-MD-231, T24 and A549), while the antioxidant potential was measured using three antioxidant methods (DPPH, FRAP and Rancimat assay). Results and conclusions. T. chamaedrys had the highest total phenolics (expressed as GAE (2061±42) mg/L), free radical scavenging activity (IC50=1.9 mg/mL) and Fe(III) reducing antioxidant power (expressed as FeCl2 (9798±27) mg/L). At a concentration of 1 mg/mL, the antiproliferation of T24 by C. majus was 96 % and of MDA-MD-231 cells by A. vulgaris was 75 % after 72 h. After principal component analysis, T. chamaedrys and C. majus were grouped together. Quercetin glucoside and antioxidant capacity (DPPH) contributed the most to differentiate these infusions from the other two. Novelty and scientific contribution. This study represents a comparative analysis of the biopotential of four medicinal plants. A new RP-HPLC method was developed to separate the flavonoids in the herbal infusions. This is the first report on the presence of kaempferol-3-O-rutinoside in C. officinalis and isorhamnetin-3-O-rutinoside in A. vulgaris aqueous infusion. For the first time, C. majus has been shown to contribute to the oxidative stability of edible oil. Furthermore, this is the first comparative study on the antiproliferative activity of selected medicinal plants against the cell lines MDA-MD-231, T24 and A549.

Alchemilla vulgaris L. is a good source of antioxidant components with an emphasis on phenolic acids and tannins. In this study, gallic acid, ellagic acid, and hydrolyzable tannins (HT) were extracted from this plant with different deep eutectic solvents (DESs), varying the amount of added H2O, temperature and extraction time. Seventeen DESs (n = 3) were used for the extraction, of which choline chloride:urea (1:2) proved to be the most suitable. The selection of the best solvent was followed by the examination of the influence of the extraction type and parameters using response surface methodology (RSM). Gallic acid content was in the range of 0.00–1.89 µg mg−1, ellagic acid content was 0.00–12.76 µg mg−1 and hydrolyzable tannin (HT) content was 3.06–181.26 µgTAE mg−1, depending on the used technique and the extraction conditions. According to the results, extraction by stirring and heating was the most suitable since the highest amounts of gallic acid, ellagic acid, and HT were extracted, and the obtained optimal values using response surface methodology (RSM) are confirmed by experimentally obtained values.