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. 2025 Jan 1;18(1):39.
doi: 10.3390/ph18010039.

Chemical Characterization and Differential Lipid-Modulating Effects of Selected Plant Extracts from Côa Valley (Portugal) in a Cell Model for Liver Steatosis

Affiliations

Chemical Characterization and Differential Lipid-Modulating Effects of Selected Plant Extracts from Côa Valley (Portugal) in a Cell Model for Liver Steatosis

Ricardo Amorim et al. Pharmaceuticals (Basel). .

Abstract

Background/objectives: Côa Valley, located in the northeast of Portugal, harbors more than 500 medicinal plant species. Among them, four species stand out due to their traditional uses: Equisetum ramosissimum Desf. (hemorrhages, urethritis, hepatitis), Rumex scutatus L. subsp. induratus (Boiss. and Reut.) Malag. (inflammation, constipation), Geranium purpureum Vill., and Geranium lucidum L. (pain relief, gastric issues). Given their rich ethnomedicinal history, we evaluated their protective effects on an in vitro model of metabolic dysfunction-associated steatotic liver disease (MASLD).

Methods: Decoction (D) and hydroalcoholic (EtOH80%) extracts were prepared and chemically characterized. Their safety profile and effects on lipid accumulation were assessed in palmitic acid (PA)-treated HepG2 cells using resazurin, sulforhodamine B, and Nile Red assays.

Results: Chemical analysis revealed diverse phenolic compounds, particularly kaempferol derivatives in E. ramosissimum. All extracts showed minimal cytotoxicity at 25-50 µg/mL. At 100 µg/mL, only E. ramosissimum extracts maintained high cell viability. In the lipotoxicity model, E. ramosissimum decoction demonstrated the most potent effect, significantly reducing PA-induced neutral lipid accumulation in a dose-dependent manner, while other extracts showed varying degrees of activity.

Conclusions: These findings highlight E. ramosissimum's decoction, rich in kaempferol derivatives, as particularly effective in reducing lipid accumulation in this MASLD cell model while also providing a comprehensive characterization of traditionally used plants from the Côa Valley region.

Keywords: Côa Valley (Portugal); Equisetum ramosissimum Desf.; MASLD; lipid-lowering effect; plant extracts.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
General aspect (A) and strobilus, cone-like structure that produces spores at the tips of a stem (B) of E. ramosissimum. Edible leaves (C) and pink-colored fruits (D) of R. induratus. Close-up on the flower, leaves (E), and ripen fruits (F) of G. purpureum. Flowers, leaves (G), and immature fruits of G. lucidum (H). Images (AD,G,H) were obtained and adapted from [28]. Images (E,F) were obtained and adapted from [29]. (I) Chemical structures of the most relevant phenolic compounds identified through HPLC–DAD–ESI/MS in E. ramosissimum, Geranium spp., and R. induratus from Côa Valley (Portugal). Chemical structures were designed in the ChemDraw Software v.14.0.
Figure 2
Figure 2
Effects of EtOH80% and decoction extracts of plants originating from the Côa Valley on cell metabolic activity. (A) Human cells study experimental timeline. The metabolic activity of HepG2 cells, in percentage of control, following extract incubation in three different concentrations (25, 50, and 100 µg/mL). Each graph contains the results for extract incubation (white bars) and for PA following extract preincubation (grey bars). The black bar represents PA at 100 µM without extract preincubation for comparison purposes. (B) EtOH80% and (C) D extract of G. purpureum, (D) EtOH80% and (E) D extract of G. lucidum, (F) EtOH80% and (G) D extract of E. ramosissimum, and (H) EtOH80% and (I) D extract of R. induratus. Statistical significance was compared using two-way ANOVA followed by Tukey post hoc test for multiple comparisons (* p < 0.05, ** p < 0.01, *** p < 0.0005, vs. untreated cells); (# p < 0.05, ## p < 0.01, ### 0.0005 vs. PA-treated cells).
Figure 3
Figure 3
Effects of EtOH80% and decoction (D) extracts of plants originating from the Côa Valley on cell mass. The cell mass of HepG2 cells, in percentage of control, following extract incubation in three different concentrations (25, 50, and 100 µg/mL). Each graph contains the results for extract incubation (white bars) and for PA following extract preincubation (grey bars). The black bar represents PA at 100 µM without extract preincubation for comparison purposes. (A) EtOH80% and (B) D extract of G. purpureum, (C) EtOH80% and (D) D extract of G. lucidum, (E) EtOH80% and (F) D extract of E. ramosissimum, (G) EtOH80% and (H) D extract of R. induratus. Statistical significance was compared using two-way ANOVA followed by Tukey post hoc test for multiple comparisons (* p < 0.05, ** p < 0.01, *** p < 0.0005, vs. untreated cells); (# p < 0.05, ## p < 0.01, #### p < 0.0001 vs. PA-treated cells).
Figure 4
Figure 4
Effects of EtOH80% and decoction extracts of plants originating from the Côa Valley on cell lipid accumulation. The neutral lipid accumulation of HepG2 cells, in percentage of control and normalized for cell mass results, following extract incubation in three different concentrations (25, 50, and 100 µg/mL). Each graph contains the results for extract incubation (white bars) and for PA following extract preincubation (grey bars). The black bar represents PA at 100 µM without extract preincubation for comparison purposes. (A) EtOH80% and (B) D extract of G. purpureum, (C) EtOH80% and (D) D extract of G. lucidum, (E) EtOH80% and (F) D extract of E. ramosissimum, (G) EtOH80% and (H) D extract of R. induratus. Statistical significance was compared using two-way ANOVA followed by Tukey post hoc test for multiple comparisons (* p < 0.05, ** p < 0.01, *** p < 0.0005, vs. untreated cells); (# p < 0.05, ## p < 0.01vs. PA-treated cells).

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