Cornus mas and Cornus officinalis-A Comparison of Antioxidant and Immunomodulatory Activities of Standardized Fruit Extracts in Human Neutrophils and Caco-2 Models

Abstract

Fruits of Cornus mas and Cornus officinalis are representative plant materials traditionally used in Europe and Asia, respectively, in the treatment of diabetes and diabetes-related complications, which are often mediated by pathogenic inflammatory agents. Additionally, due to the fact of mutual infiltration of Asian and European medicines, the differentiation as well as standardization of traditional prescriptions seem to be crucial for ensuring the quality of traditional products. The objective of this study was a comparison of biological activity of extracts from fruits of C. mas and C. officinalis by an assessment of their effect on reactive oxygen species (ROS) generation in human neutrophils as well as cytokines secretion both in neutrophils (tumor necrosis factor α, TNF- α; interleukin 8, IL-8; interleukin 1β, IL-1β) and in human colon adenocarcinoma cell line Caco-2 (IL-8). To evaluate the phytochemical differences between the studied extracts as well as to provide a method for standardization procedures, a quantitative analysis of iridoids, such as loganin, sweroside, and loganic acid, found in extracts of Cornus fruits was performed with HPLC-DAD. All standardized extracts significantly inhibited ROS production, whereas the aqueous-alcoholic extracts were particularly active inhibitors of IL-8 secretion by neutrophils. The aqueous-methanolic extract of C. officinalis fruit, decreased IL-8 secretion by neutrophils to 54.64 ± 7.67%, 49.68 ± 6.55%, 50.29 ± 5.87% at concentrations of 5, 50, and 100 µg/mL, respectively, compared to LPS-stimulated control (100%). The aqueous extract of C. officinalis fruit significantly inhibited TNF-α release by neutrophils at concentrations of 50 and 100 µg/mL. On the other hand, the aqueous-ethanolic extract of C. mas fruit showed the propensity to increase TNF-α and IL-1β secretion. The modulatory activity of the Cornus extracts was noted in the case of secretion of IL-8 in Caco-2 cells. The effect was comparable with dexamethasone. The content of loganin in aqueous and aqueous-methanolic extract of C. officinalis fruit was higher than in the aqueous-ethanolic extract of C. mas fruit, which was characterized by a significant quantity of loganic acid. In conclusion, the immunomodulatory effect observed in vitro may partially confirm the traditional use of Cornus fruits through alleviation of the development of diabetes-derived inflammatory complications. Loganin and loganic acid are significant markers for standardization of C. mas and C. officinalis fruit extracts, respectively.

Keywords: dogwood; inflammation; loganic acid; loganin; traditional medicine.

Figure 1

f-MLP-stimulated ROS production by PMNs (mean ± SEM [%]) after the treatment by: (a) Cornus extracts; (b) standard compounds. CM_et—aqueous ethanolic extract of Cornus mas fruit, CO_met—aqueous-methanolic extract of Cornus officinalis fruit, CO_aq—aqueous extract of Cornus officinalis fruit; ^# p < 0.001 vs. (−) LPS; * p < 0.05, ** p < 0.001 vs. (+) LPS.

Figure 2

Effect of Cornus extracts on LPS-stimulated IL-8 secretion by PMNs (mean ± SEM [%]). CM_et—aqueous ethanolic extract of Cornus mas fruit, CO_met—aqueous-methanolic extract of Cornus officinalis fruit, CO_aq–aqueous extract of Cornus officinalis fruit, Dex—dexamethasone; ^# p < 0.001 vs. (−) LPS; * p < 0.05, ** p < 0.001 vs. (+) LPS.

Figure 3

Effect of Cornus extracts on LPS-stimulated TNF-α secretion by PMNs (mean ± SEM [%]). CM_et—aqueous ethanolic extract of Cornus mas fruit, CO_met—aqueous-methanolic extract of Cornus officinalis fruit, CO_aq—aqueous extract of Cornus officinalis fruit, Dex—dexamethasone; ^# p < 0.001 vs. (−) LPS; * p < 0.05, ** p < 0.001 vs. (+) LPS.

Figure 4

Effect of Cornus extracts on LPS-stimulated IL-1β secretion by PMNs (mean ± SEM [%]). CM_et—aqueous ethanolic extract of Cornus mas fruit, CO_met—aqueous-methanolic extract of Cornus officinalis fruit, CO_aq—aqueous extract of Cornus officinalis fruit, Dex—dexamethasone; ^# p < 0.001 vs. (−) LPS; * p < 0.05, ** p < 0.001 vs. (+) LPS.

Figure 5

Effect of Cornus extracts on IL-8 secretion by Caco-2 (mean ± SEM [%]). CM_et—aqueous ethanolic extract of Cornus mas fruit, CO_met—aqueous-methanolic extract of Cornus officinalis fruit, CO_aq—aqueous extract of Cornus officinalis fruit, Dex—dexamethasone, st.—mixture of IL-1β/TNF-α/IFN-γ/LPS; ^# p < 0.001 vs. (−) LPS; * p < 0.05 vs. (+) LPS.

Figure 6

Effect of Cornus extracts on cells viability (a) PMNs; (b) Caco-2 cells. Data are expressed mean ± SEM [%]. Control n.t.—non-treated control cells; ^# p < 0.001 vs. control n.t.

Figure 7

HPLC chromatograms registered at 240 nm (a) aqueous-ethanolic extract of Cornus mas fruit; (b) aqueous-methanolic extract of Cornus officinalis fruit; (c) aqueous extract of Cornus officinalis fruit. Loganic acid (1), loganin (2), sweroside (3).