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Keywords = Nepeta nuda L.

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23 pages, 9939 KB  
Article
Extraction Temperatures Shape Water-Soluble Metabolite Profiles of Nepeta nuda L. and thus Modulate the Bioactive Properties
by Desislava Mantovska, Alexandra Kapogianni, Ginka Cholakova, Ivanka Tsacheva, Anton Hinkov, Detelina Petrova, Zlatina Gospodinova, Georgi Antov, Danijela Mišić, Krasimir Rusanov, Mila Rusanova, Kalina Shishkova, Momchil Paunov, Zhenya Yordanova and Miroslava Zhiponova
Metabolites 2026, 16(5), 323; https://doi.org/10.3390/metabo16050323 - 13 May 2026
Viewed by 721
Abstract
Background: Plants of the genus Nepeta are widely used in ethnomedicine for treating inflammatory disorders due to their rich content of bioactive compounds. This study investigated how extraction temperature specifically affects the bioactive potential of aqueous extracts from wild-grown Nepeta nuda L. Methods: [...] Read more.
Background: Plants of the genus Nepeta are widely used in ethnomedicine for treating inflammatory disorders due to their rich content of bioactive compounds. This study investigated how extraction temperature specifically affects the bioactive potential of aqueous extracts from wild-grown Nepeta nuda L. Methods: The previously used maceration approach for this plant was applied at 30–60 °C to flowers, leaves, and stems. Phytochemical profiling included spectrophotometric assays, metabolite identification, and quantification. Biological activities reported for this plant were assessed, including antioxidant, anti-inflammatory, antiviral, antiproliferative, and antibacterial capacities. Results: Extraction yield was highest in flowers and leaves, where it increased significantly with rising temperature, while stems were less productive. All plant organs exhibited notable bioactivity falling into two groups: lower temperatures (30 and 40 °C) were optimal for antiviral and anti-inflammatory effects, whereas and higher temperatures (50 and 60 °C) enhanced antioxidant potential. The phytochemical composition, evaluated at representative extraction temperatures, revealed differential accumulation of p-coumaric acid and luteolin in all organs at 40 °C, while extraction at 60 °C corresponded to elevated levels of phenolic compounds. Flower extracts were confirmed to have the richest metabolic composition and were therefore subjected to further investigation. Extracts obtained at 40 °C influenced C1q binding, supporting their anti-inflammatory activity, whereas extraction at 60 °C resulted in stronger antiproliferative activity in colon cancer cell line. Antibacterial effects were similar at both temperatures. Conclusions: These findings highlight the importance of optimizing extraction conditions for future pharmacological applications of N. nuda. Full article
(This article belongs to the Special Issue Bioactive Metabolites from Natural Sources (2nd Edition))
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21 pages, 3109 KB  
Article
Salicylic Acid-Induced Elicitation of Nepetalactone and Rosmarinic Acid Biosynthesis in Naked Catmint (Nepeta nuda L.): Metabolomic and Transcriptional Insights
by Luka Petrović, Slavica Dmitrović, Jasmina Nestorović Živković, Biljana Filipović, Neda Popović, Milica Milutinović, Dragana Matekalo, Uroš Gašić, Danijela Mišić and Marijana Skorić
Int. J. Mol. Sci. 2026, 27(8), 3570; https://doi.org/10.3390/ijms27083570 - 16 Apr 2026
Cited by 1 | Viewed by 582
Abstract
Salicylic acid (SA) is involved in plant defense responses to environmental stressors by modulating gene expression and specialized metabolites production, enhancing plant adaptive resilience through systemic signaling pathways. This study investigates the impact of exogenous application of SA on the metabolism of iridoids [...] Read more.
Salicylic acid (SA) is involved in plant defense responses to environmental stressors by modulating gene expression and specialized metabolites production, enhancing plant adaptive resilience through systemic signaling pathways. This study investigates the impact of exogenous application of SA on the metabolism of iridoids and phenolic compounds—characteristic specialized metabolites of the Nepeta species, associated with diverse biological activities. Nepetalactone (NL) is a characteristic monoterpene iridoid, while rosmarinic acid (RA) represents the most abundant phenolic compound within the genus. We explored the effects of varying SA concentrations (2 µM, 5 µM, 10 µM, and 20 µM) on iridoid and phenolic metabolism in in vitro-grown Nepeta nuda, following 7 days and 28 days of elicitation. A significant increase in trans,trans-NL content was observed after 7-day exposure to 2 µM SA, while prolonged exposure led to a decrease in its levels, particularly at higher SA concentrations. Gene expression analysis revealed that 7 days of exposure to lower concentrations of SA upregulated genes coding for NAD-dependent nepetalactol-related short-chain dehydrogenase/reductases (NEPSs), key regulatory enzymes catalyzing the final steps of NL biosynthesis. In contrast, prolonged exposure to 20 µM SA downregulated genes coding for geraniol 8-hydroxylase (NnG8H) and 8-hydroxygeraniol oxidoreductase (Nn8HGO), which resulted in reduced iridoid content. Conversely, SA treatment notably increased RA content after prolonged exposure to 20 µM SA, which is a result of the enhanced expression of all analyzed RA biosynthesis-related genes. These findings demonstrate that both concentration and duration of SA treatment are critical determinants of elicitation outcomes in N. nuda. Strategic manipulation of these parameters can redirect metabolic flux toward either iridoid or phenolic compounds production, and enhance biotechnological production of specialized metabolites in N. nuda. Full article
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16 pages, 3926 KB  
Article
In Vivo Assessment of Anti-Inflammatory Effects of Aqueous Extracts of Nepeta nuda ssp. nuda L. in Experimental Model of Peripheral Inflammation in Male Long Evans Rats
by Milena Keremidarska-Markova, Veneta Evtimova-Koeva, Tsvetozar Penchev, Dilyana Doncheva-Stoimenova, Miroslava Zhiponova, Mariela Chichova and Bilyana Ilieva
Life 2025, 15(12), 1938; https://doi.org/10.3390/life15121938 - 18 Dec 2025
Cited by 2 | Viewed by 774
Abstract
Background: Recently various plants have attracted considerable scientific interest as potential therapeutic alternatives to known drugs used in anti-inflammatory therapy. Therefore, we have investigated the possible anti-inflammatory and analgesic effects of aqueous extracts from flowers of the medical plant Nepeta nuda ssp. [...] Read more.
Background: Recently various plants have attracted considerable scientific interest as potential therapeutic alternatives to known drugs used in anti-inflammatory therapy. Therefore, we have investigated the possible anti-inflammatory and analgesic effects of aqueous extracts from flowers of the medical plant Nepeta nuda ssp. nuda L. (naked catmint) in a model of acute peripheral inflammation induced by intraplantar injection of λ-carrageenan in the hind paw of Long Evans rats. Methods: Two routes of N. nuda extract application were used: locally by intraplantar injections at dosages of 2.5 mg/kg and 25 mg/kg and systemically by intraperitoneal administration at dosages of 50 mg/kg and 200 mg/kg, respectively. Paw volume was measured prior to the carrageenan application and 1, 2, 3, and 4 h after carrageenan injection. Spontaneous locomotor activity of the rats was assessed 3 h after carrageenan injection, corresponding to the peak of acute paw inflammation. Results: Local application of the higher N. nuda dose led to a marked reduction in inflammatory paw edema at the 4th hour after carrageenan injection, comparable to the effect of the positive control diclofenac sodium. Interestingly, a similar anti-inflammatory effect was observed at the 1st hour when both extract doses were administered intraperitoneally. Only the higher intraplantar dose of N. nuda extract significantly enhanced the vertical activity in comparison to the group treated with carrageenan alone. Conclusions: Our data indicate that the aqueous N. nuda extract possesses potent anti-inflammatory effects following both local and systemic administration in rats. Furthermore, when administered locally the extract exerts significant analgesic activity in inflammatory pain. Full article
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17 pages, 1941 KB  
Article
Blue–Red LED Light Modulates Morphophysiological and Metabolic Responses in the Medicinal Plant Nepeta nuda
by Miroslava Zhiponova, Grigor Zehirov, Krasimir Rusanov, Mila Rusanova, Miroslava Stefanova, Tsveta Ganeva, Momchil Paunov, Valentina Ganeva, Kiril Mishev, Petre I. Dobrev, Roberta Vaculíková, Václav Motyka, Zhenya Yordanova, Ganka Chaneva and Valya Vassileva
Plants 2025, 14(15), 2285; https://doi.org/10.3390/plants14152285 - 24 Jul 2025
Cited by 2 | Viewed by 1347
Abstract
Light quality and duration profoundly influence the growth and productivity of plant species. This study investigated the effects of a blue–red LED light combination, known to induce flowering, on the physiological state and content of biologically active substances in catmint (Nepeta nuda [...] Read more.
Light quality and duration profoundly influence the growth and productivity of plant species. This study investigated the effects of a blue–red LED light combination, known to induce flowering, on the physiological state and content of biologically active substances in catmint (Nepeta nuda L.) grown under controlled in vitro conditions. White light (W) was used as a control and compared with two blue–red intensities: BR (high-intensity blue–red light) and BRS (low-intensity blue–red light or “BR with shadow”). BR-treated plants showed increased leaf area, mesophyll thickness, biomass and starch content but reduced levels of plastid pigments. BR also modified the oxidative state of plants by inducing lipid peroxidation while simultaneously activating ROS scavenging mechanisms and enhancing phenolic antioxidants. Interestingly, BR decreased the accumulation of the Nepeta sp.-specific iridoid, nepetalactone. These effects appear to be regulated by the phytohormones auxin, abscisic acid and jasmonates. BRS treatment produced effects similar to the W control but led to increased plant height and reduced leaf area and thickness. Both BR and BRS regimes induced the accumulation of proteins and amino acids. We conclude that blue–red light can enhance the survival capacity of micropropagated N. nuda during subsequent soil adaptation, suggesting that similar light pre-treatment could improve plant performance under stress conditions. Full article
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14 pages, 2642 KB  
Article
In Vitro Anticancer Effects of Aqueous Leaf Extract from Nepeta nuda L. ssp. nuda
by Zlatina Gospodinova, Georgi Antov, Svetozar Stoichev and Miroslava Zhiponova
Life 2024, 14(12), 1539; https://doi.org/10.3390/life14121539 - 24 Nov 2024
Cited by 9 | Viewed by 1662
Abstract
Despite significant efforts, cancer remains the second leading cause of mortality worldwide. The medicinal plant Nepeta nuda L. represents a valuable source of biologically active compounds with pharmacological activities including antioxidant, anti-inflammatory, antimicrobial, and antiviral. This study aimed to assess the antiproliferative potential [...] Read more.
Despite significant efforts, cancer remains the second leading cause of mortality worldwide. The medicinal plant Nepeta nuda L. represents a valuable source of biologically active compounds with pharmacological activities including antioxidant, anti-inflammatory, antimicrobial, and antiviral. This study aimed to assess the antiproliferative potential and mechanisms of action of aqueous extract from the leaves of wild-grown N. nuda. Cancer cell lines, MDA-MB-231, MCF7 (breast), HT29, Colon 26 (colon), and HepG2 (liver cancer), and a non-cancerous skin cell line, BJ, were assessed for antiproliferative activity by MTT assay and observation of cell morphological alterations. The cancer cell line that was most sensitive to the extract was further studied for apoptotic alterations by Annexin V/propidium iodide staining, colony-forming assay, and qRT-PCR analysis. The results revealed that the plant extract inhibited the proliferation of all investigated cancer cell lines with the strongest cytostatic effect on Colon 26 cells with a half maximal inhibitory concentration (IC50) value of 380.2 μg/mL and a selectivity index (SI) of 3.5. The extract significantly inhibited the ability of cells to form colonies, exhibited considerable proapoptotic potential involving the participation of the CASP8 gene, and increased the expression levels of ATG3 and the BECN1 gene, which suggests a role of autophagic cell death in the antitumor action. Full article
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29 pages, 10649 KB  
Article
Patterns of Genetic Variation of Nepeta nuda L. from the Central Balkans: Understanding Drivers of Chemical Diversity
by Luka Petrović, Marijana Skorić, Branislav Šiler, Tijana Banjanac, Uroš Gašić, Dragana Matekalo, Tamara Lukić, Jasmina Nestorović Živković, Slavica Dmitrović, Neda Aničić, Milica Milutinović, Jelena Božunović, Biljana Filipović, Miloš Todorović and Danijela Mišić
Plants 2024, 13(11), 1483; https://doi.org/10.3390/plants13111483 - 27 May 2024
Cited by 11 | Viewed by 2678
Abstract
Nepeta nuda L., a notable medicinal species in the tradition of the Balkan region, is a rich source of bioactive iridoids and phenolics previously described as high-resolution taxonomical classifiers for the genus Nepeta. However, their potential in investigating intra-species differentiation is here [...] Read more.
Nepeta nuda L., a notable medicinal species in the tradition of the Balkan region, is a rich source of bioactive iridoids and phenolics previously described as high-resolution taxonomical classifiers for the genus Nepeta. However, their potential in investigating intra-species differentiation is here described for the first time. The aim was to recognize the sources of natural chemical diversity and their association with the genetic variability both within and among N. nuda populations in the Central Balkans. Chemical diversity was assessed from methanol extracts and essential oils through untargeted and targeted metabolomics using state-of-the-art analytical tools, covering a broad spectrum of compounds that represent the N. nuda metabolome. We found that chemodiversity primarily resides within populations of N. nuda, and similar results were obtained at the DNA level using microsatellite markers. The low genetic and chemical differentiation of the studied N. nuda populations implies that their metabolomic profiles may be less influenced by geographic distance and variable environmental conditions within the Central Balkans, as they are under the pivotal control of their genetic backgrounds. Screening the distribution of the major bioactive compounds belonging to phenolics (phenolic acids and flavonoids) and iridoids (both aglycones and glycosylated forms), within and among N. nuda populations, is able to guarantee mass spectrometry-based tools for the selection of elite representative genotypes with practical importance. The knowledge acquired will allow us to delve deeper into the molecular background of N. nuda chemical diversity, which is the course of our further work. Full article
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18 pages, 7328 KB  
Article
Uncovering the Interrelation between Metabolite Profiles and Bioactivity of In Vitro- and Wild-Grown Catmint (Nepeta nuda L.)
by Anna Zaharieva, Krasimir Rusanov, Mila Rusanova, Momchil Paunov, Zhenya Yordanova, Desislava Mantovska, Ivanka Tsacheva, Detelina Petrova, Kiril Mishev, Petre I. Dobrev, Jozef Lacek, Roberta Filepová, Grigor Zehirov, Valya Vassileva, Danijela Mišić, Václav Motyka, Ganka Chaneva and Miroslava Zhiponova
Metabolites 2023, 13(10), 1099; https://doi.org/10.3390/metabo13101099 - 20 Oct 2023
Cited by 18 | Viewed by 3311
Abstract
Nepeta nuda L. is a medicinal plant enriched with secondary metabolites serving to attract pollinators and deter herbivores. Phenolics and iridoids of N. nuda have been extensively investigated because of their beneficial impacts on human health. This study explores the chemical profiles of [...] Read more.
Nepeta nuda L. is a medicinal plant enriched with secondary metabolites serving to attract pollinators and deter herbivores. Phenolics and iridoids of N. nuda have been extensively investigated because of their beneficial impacts on human health. This study explores the chemical profiles of in vitro shoots and wild-grown N. nuda plants (flowers and leaves) through metabolomic analysis utilizing gas chromatography and mass spectrometry (GC–MS). Initially, we examined the differences in the volatiles’ composition in in vitro-cultivated shoots comparing them with flowers and leaves from plants growing in natural environment. The characteristic iridoid 4a-α,7-β,7a-α-nepetalactone was highly represented in shoots of in vitro plants and in flowers of plants from nature populations, whereas most of the monoterpenes were abundant in leaves of wild-grown plants. The known in vitro biological activities encompassing antioxidant, antiviral, antibacterial potentials alongside the newly assessed anti-inflammatory effects exhibited consistent associations with the total content of phenolics, reducing sugars, and the identified metabolic profiles in polar (organic acids, amino acids, alcohols, sugars, phenolics) and non-polar (fatty acids, alkanes, sterols) fractions. Phytohormonal levels were also quantified to infer the regulatory pathways governing phytochemical production. The overall dataset highlighted compounds with the potential to contribute to N. nuda bioactivity. Full article
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4 pages, 1010 KB  
Proceeding Paper
Nepeta nuda L. Plant Extract Preserves the Morphology of Red Blood Cells Subjected to Oxidative Stress
by Stefani Petrova, Nikol Mazhdrakova, Svetla Todinova, Velichka Strijkova, Miroslava Zhiponova and Sashka Krumova
Med. Sci. Forum 2023, 21(1), 35; https://doi.org/10.3390/ECB2023-14086 - 1 Mar 2023
Cited by 2 | Viewed by 1878
Abstract
We show that an extract from catmint (Nepeta nuda L.) flowers is not hemotoxic and does not alter erythrocytes’ morphology. H2O2-induced oxidative stress leads to an increase in lipid peroxidation, accompanied by a reduction in the number of [...] Read more.
We show that an extract from catmint (Nepeta nuda L.) flowers is not hemotoxic and does not alter erythrocytes’ morphology. H2O2-induced oxidative stress leads to an increase in lipid peroxidation, accompanied by a reduction in the number of biconcave cells and an increase in the number of echinocytes. Pre-treatment of erythrocytes with this extract does not reduce the lipid peroxidation level; however, it results in partial restoration of the relative abundance of biconcave cells and a respective reduction in the echinocytes’ quantity. Our data reveal the concentrations at which the examined extract exhibits a protective effect on erythrocytes’ morphology under the condition of H2O2-induced oxidative stress. Full article
(This article belongs to the Proceedings of The 2nd International Electronic Conference on Biomedicines)
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13 pages, 2378 KB  
Article
Volatile Compounds of Nepeta nuda L. from Rtanj Mountain (Serbia)
by Milica Aćimović, Biljana Lončar, Milada Pezo, Jovana Stanković Jeremić, Mirjana Cvetković, Milica Rat and Lato Pezo
Horticulturae 2022, 8(2), 85; https://doi.org/10.3390/horticulturae8020085 - 18 Jan 2022
Cited by 20 | Viewed by 4085
Abstract
Unlike other studies that only determined the chemical composition of essential oils depending on their geographic origin, this research investigated the effect of weather conditions (temperature, precipitation, and insolation) on the chemical composition of Nepeta nuda L. essential oil. The collection of wild-growing [...] Read more.
Unlike other studies that only determined the chemical composition of essential oils depending on their geographic origin, this research investigated the effect of weather conditions (temperature, precipitation, and insolation) on the chemical composition of Nepeta nuda L. essential oil. The collection of wild-growing N. nuda was carried out during three successive years, 2019, 2020, and 2021 at Rtanj Mountain (Serbia) on the same date (July 7th). Essential oil extraction from the plant was performed by hydro-distillation. After gas chromatographic-mass spectrometric analysis, a total of 102 volatile compounds were separated from N. nuda, during the observed period, 28 were unidentified, compromising between 5.0% and 8.7%, depending on the year. A multiple linear regression model was created, and statistical analyses were performed to provide knowledge about the prediction, feature profile, and the similarity in contents of active compounds of the N. nuda essential oil. The influence of temperature on the accumulation of the most abundant component, 1,8-cineole, was positive, while the impact of precipitation and insolation was negative. According to the cluster tree, there are four chemotypes of N. nuda essential oil: with nepetalactone, 1,8-cineole, mixed (nepetalactone+1,8-cineole+germacrene D), and nonspecific chemotypes. Bearing in mind that the biological activity of a raw material depends on the chemotype and environmental factors, this is a topic that deserves a more detailed approach. The N. nuda and its essential oil are promising materials with high biological potential, and these deserve further detailed investigation. Full article
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