Oxydative stress, anxiety and depression are associated with changes in cytokine levels. Natural products, including individual and combined plant extracts, have the potential to be used in the treatment of neuropsychiatric disorders. The goal of this study is to investigate the effects of two combined plant extracts, rich in flavonoids, on the levels of the cytokines TNF-α, IL-6, and IL-10 in rats subjected to models of acute cold stress and chronic unpredictable stress. The study utilized common medicinal plants such as Valeriana officinalis, Melissa officinalis, Crataegus monogyna, Hypericum perforatum, and Serratula coronata, which were combined in two unique combinations—Antistress I and Antistress II. The compositions of the used extracts were determined by HPLC methods. Pro- and anti-inflammatory cytokines in rats’ serum were measured with Enzyme-linked immunosorbent assay. The results from the acute stress model revealed that the individual extract of Crataegus monogyna decreased levels of TNF-α, while Serratula coronata, Hypericum perforatum, and Valeriana officinalis effectively reduced IL-6 levels. Both combinations, Antistress I and Antistress II, were effective in reducing TNF-α and IL-6 levels, with Antistress II also increasing IL-10 levels. In the chronic stress model, Hypericum perforatum extract decreased the levels of the pro-inflammatory cytokines TNF-α and IL-6, whereas extracts of Serratula coronata and Valeriana officinalis only reduced TNF-α levels. The two combined extracts, Antistress I and Antistress II, decreased TNF-α and IL-6 levels, while Antistress I also reduced the levels of the anti-inflammatory cytokine IL-10. The combinations of plant extracts used in our experiment have not been previously studied or documented in the available literature. However, based on our own experimental results, we can draw the conclusion that the combinations exhibit a more pronounced effect in reducing cytokine levels compared to the individual plant extracts.

Cannabidiol (CBD), a non-psychoactive compound derived from Cannabis sativa, is believed to have anxiety-reducing and antidepressant effects. However, existing data are inconsistent, likely due to variations in experimental designs, dosages, and stress models. This study sought to assess the impact of CBD on anxiety and depression-like behaviors in Wistar rats exposed to acute cold stress, as well as its impact on pro- and anti-inflammatory cytokines. Male rats were treated with CBD (2.5, 5, or 10 mg/kg) or vehicle for 14 days and subjected to behavioral tests, including the elevated plus maze, social interaction, and forced swim tests. Serum levels of cytokines (IL-6, TNF-α, IL-1β, and IL-10) were analyzed post-experiment using ELISA. Results demonstrated a dose-dependent anxiolytic effect of CBD, with significant improvements in social interaction and reductions in anxiety-like behaviors at 5 and 10 mg/kg. All doses of CBD decreased immobility in the forced swim test, suggesting antidepressant effects. Furthermore, CBD selectively lowered IL-6 levels, a key cytokine in acute stress and depression pathogenesis. These findings indicate that CBD has anxiety-reducing and antidepressant properties, partially mediated by modulation of inflammatory processes, particularly IL-6.

Echinacea purpurea and Onopordum acanthium, which belong to the Asteraceae family, are widely used plants in traditional medicine. Their antioxidant, anti-inflammatory, antiviral, antibacterial, and antitumor effects are well known. However, there are no data on the effects of their combination. The aim of the present study was to combine E. purpurea with O. acanthium to study the in vivo immunomodulatory effect of two combinations and to compare it with that of single plants. Their total polyphenolic and flavonoid content and the amounts of individual compounds characteristic of both species were determined. The influence of the obtained extracts on the serum concentrations of cytokines IFN-γ, TNF-α, and IL-10 in experimental animals with lipopolysaccharide-induced systemic inflammatory response was investigated. This research found that a combination of E. purpurea/O. acanthium in the ratio 1:1 reduced the proinflammatory cytokines TNF-α (244.82 pg/mL) and IFN-γ (1327.92 pg/mL) compared to the LPS-control, respectively, (574.17 pg/mL) and (3354.00 pg/mL), and the combination E. purpurea/O. acanthium in the ratio of 3:1 significantly increased the levels of the anti-inflammatory cytokine IL-10 (1313.95 pg/mL) compared to the LPS-control (760.09 pg/mL). In conclusion, our results could be a basis for future biomedical research on creating phytopreparations with an immunomodulatory effect.

Background: Echinacea purpurea (L.) Moench (EP) and Onopordum acanthium (L.) (OA) are promising medicinal plants with diverse biological activities but there is no information on the effects of their combinations. To harness the therapeutic potential of both while minimizing the risk of adverse effects, we prepared two combinations (CE1 and CE2) of EP and OA in ratios 1:1 and 3:1, respectively. Methods: Oxygen radical absorbance capacity (ORAC), hydroxyl radical absorbance capacity (HORAC), and an electrochemical assay were used to determine the antioxidant activity of the extracts in vitro. The anxiolytic and immunomodulatory properties were studied in rats. Animals were subjected to acute cold stress and anxiety-like behavior was evaluated by the elevated plus maze (EPM) and social interaction test (SIT). Serum IFN-γ, TNF-α and IL-10 levels were measured by ELISA. Results: CE2 demonstrated the highest antioxidant activity (1841.7 μmolTE/g by ORAC, 277.2 GAE/g by HORAC, and 39.6 by electrochemical method). Moreover CE2 produced anxiolytic-like effects—significantly increasing the open arms entries ratio (OAER; p < 0.001), open arms time ratio (OATR; p < 0.01) in the EPM, and prolonging the social interaction time (p < 0.05) versus the stressed control. OA increased OAER (p < 0.01) and OATR (p < 0.001), while EP increased only OAER (p < 0.01). CE1 showed no significant behavioral consequences. CE2 significantly reduced IFN-γ (p < 0.05), and IL-10 levels were elevated in OA and CE2 groups (p < 0.01). No significant changes in TNF-α levels were observed across groups. Conclusions: These findings indicate that CE2 and OA attenuate anxiety-like behavior and modulate the immune response primarily by stimulating IL-10 production.