Ethanolic Extract of Propolis and CAPE as Cardioprotective Agents against LPS and IFN-α Stressed Cardiovascular Injury.

The inflammatory process is triggered by several factors such as toxins, pathogens, and damaged cells, promoting inflammation in various systems, including the cardiovascular system, leading to heart failure. The link between periodontitis as a chronic inflammatory disease and cardiovascular disease is confirmed. Propolis and its major component, caffeic acid phenethyl ester (CAPE), exhibit protective mechanisms and anti-inflammatory effects on the cardiovascular system. The objective of the conducted study was to assess the anti-inflammatory effects of the Polish ethanolic extract of propolis (EEP) and its major component-CAPE-in interferon-alpha (IFN-α), lipopolysaccharide (LPS), LPS + IFN-α-induced human gingival fibroblasts (HGF-1). EEP and CAPE were used at 10-100 µg/mL. A multiplex assay was used for interleukin and adhesive molecule detection. Our results demonstrate that EEP, at a concentration of 25 µg/mL, decreases pro-inflammatory cytokine IL-6 in LPS-induced HGF-1. At the same concentration, EEP increases the level of anti-inflammatory cytokine IL-10 in LPS + IFN-α-induced HGF-1. In the case of CAPE, IL-6 in LPS and LPS + IFN-α induced HGF-1 was decreased in all concentrations. However, in the case of IL-10, CAPE causes the highest increase at 50 µg/mL in IFN-α induced HGF-1. Regarding the impact of EEP on adhesion molecules, there was a noticeable reduction of E-selectin by EEP at 25, 50, and100 µg/mL in IFN-α -induced HGF-1. In a range of 10-100 µg/mL, EEP decreased endothelin-1 (ET-1) during all stimulations. CAPE statistically significantly decreases the level of ET-1 at 25-100 µg/mL in IFN-α and LPS + IFN-α. In the case of intercellular adhesion molecule-1 (ICAM-1), EEP and CAPE downregulated its expression in a non-statistically significant manner. Based on the obtained results, EEP and CAPE may generate beneficial cardiovascular effects by influencing selected factors. EEP and CAPE exert an impact on cytokines in a dose-dependent manner.

In Vitro Anti-Inflammatory Activity of Methyl Derivatives of Flavanone.

Inflammation plays an important role in the immune defense against injury and infection agents. However, the inflammatory chronic process may lead to neurodegenerative diseases, atherosclerosis, inflammatory bowel diseases, or cancer. Flavanones present in citrus fruits exhibit biological activities, including anti-oxidative and anti-inflammatory properties. The beneficial effects of flavanones have been found based on in vitro cell cultures and animal studies. A suitable in vitro model for studying the inflammatory process are macrophages (RAW264.7 cell line) because, after stimulation using lipopolysaccharide (LPS), they release inflammatory cytokines involved in the immune response. We determined the nitrite concentration in the macrophage cell culture and detected ROS using chemiluminescence. Additionally, we measured the production of selected cytokines using the Bio-Plex Magnetic Luminex Assay and the Bio-PlexTM 200 System. For the first time, we have shown that methyl derivatives of flavanone inhibit NO and chemiluminescence generated via LPS-stimulated macrophages. Moreover, the tested compounds at 1-20 µM dose-dependently modulate proinflammatory cytokine production (IL-1ß, IL-6, IL-12p40, IL-12p70, and TNF-α) in stimulated RAW264.7 cells. The 2'-methylflavanone (5B) and the 3'-methylflavanone (6B) possess the strongest anti-inflammatory activity among all the tested flavanone derivatives. These compounds reduce the concentration of IL-6, IL-12p40, and IL12p70 compared to the core flavanone structure. Moreover, 2'-methylflavanone reduces TNF-α, and 3'-methylflavanone reduces IL-1ß secreted by RAW264.7 cells.

The Phenolic Profile and Anti-Inflammatory Effect of Ethanolic Extract of Polish Propolis on Activated Human Gingival Fibroblasts-1 Cell Line.

Propolis, owing to its antibacterial and anti-inflammatory properties, acts as a cariostatic agent, capable of preventing the accumulation of dental plaque and inhibiting inflammation. The anti-inflammatory properties of propolis are attributed to caffeic acid phenethyl ester (CAPE), which is present in European propolis. The objective of the conducted study was to assess the anti-inflammatory effects of the Polish ethanolic extract of propolis (EEP) and isolated CAPE on stimulated with LPS and IFN-α, as well as the combination of LPS and IFN-α. The cytotoxicity of the tested compounds was determined using the MTT assay. The concentrations of specific cytokines released by the HGF-1 cell line following treatment with EEP (25-50 µg/mL) or CAPE (25-50 µg/mL) were assessed in the culture supernatant. In the tested concentrations, both CAPE and EEP did not exert cytotoxic effects. Our results demonstrate that CAPE reduces TNF-α and IL-6 in contrast to EEP. Propolis seems effective in stimulating HGF-1 to release IL-6 and IL-8. A statistically significant difference was observed for IL-8 in HGF-1 stimulated by LPS+IFN-α and treated EEP at a concentration of 50 µg/mL (p = 0.021201). Moreover, we observed that CAPE demonstrates a stronger interaction with IL-8 compared to EEP, especially when CAPE was administered at a concentration of 50 µg/mL after LPS + IFN-α stimulation (p = 0.0005). Analysis of the phenolic profile performed by high-performance liquid chromatography allowed identification and quantification in the EEP sample of six phenolic acids, five flavonoids, and one aromatic ester-CAPE. Propolis and its compound-CAPE-exhibit immunomodulatory properties that influence the inflammatory process. Further studies may contribute to explaining the immunomodulatory action of EEP and CAPE and bring comprehensive conclusions.

Santin (5,7-Dihydroxy-3,6,4'-Trimetoxy-Flavone) Enhances TRAIL-Mediated Apoptosis in Colon Cancer Cells.

TRAIL (Tumor necrosis factor-Related Apoptosis-Inducing Ligand) has the ability to selectively kill cancer cells without being toxic to normal cells. This endogenous ligand plays an important role in surveillance and anti-tumor immunity. However, numerous tumor cells are resistant to TRAIL-induced apoptosis. In this study, the apoptotic effect of santin in combination with TRAIL on colon cancer cells was examined. Flow cytometry was used to detect the apoptosis and expression of death receptors (TRAIL-R1/DR4 and TRAIL-R2/DR5). Mitochondrial membrane potential (ΔΨm) was evaluated by DePsipher staining with the use of fluorescence microscopy. We have shown for the first time that flavonoid santin synergizes with TRAIL to induce apoptosis in colon cancer cells. Santin induced TRAIL-mediated apoptosis through increased expression of death receptors TRAIL-R1 and TRAIL-R2 and augmented disruption of the mitochondrial membrane in SW480 and SW620 cancer cells. The obtained data may indicate the potential role of santin in colon cancer chemoprevention through the enhancement of TRAIL-mediated apoptosis.

[Apigenin inhibits release and gene expression of monocyte chemoattractant protein 1 (MCP-1) in J774.2 macrophages]. / Hamujacy wplyw apigeniny na wydzielanie i ekspresji genu bialka chemotaktycznego monocytów (MCP-1) w hodowli makrofagów linii J774.2.

Monocyte chemoattractant protein-1(MCP-1) is secreted by activated macrophages and endothelial cells, and is involved in the pathogenesis of cardiovascular and neurodegenerative disorders. There is need to develop drugs that inhibit excessive infiltration of monocytes and lymphocytes to the arterial wall and central nervous system. The aim of this study was to evaluate the effect of apigenin on the synthesis and release of MCP-1 by J774.2 macrophages in vitro. Apigenin studied at higher doses (10 and 30 microM) diminished MCP-1 release in lipopolysaccharide activated J774.2 macrophages. Apigenin administered at lower doses (3.1 and 0.3 microM) did not change secretion of MCP-1 in LPS-stimulated macrophages. Apigenin treated at a dose of 30 microM strongly reduced the number of MCP-1 mRNA copies in J774.2 cells. These results suggest that apigenin possesses anti-inflammatory properties and that apigenin inhibited MCP-1 production at the transcriptional level.

Effect of apigenin, kaempferol and resveratrol on the expression of interleukin-1beta and tumor necrosis factor-alpha genes in J774.2 macrophages.

Flavonoids have been reported to bring benefits in lowering inflammation, oxidative stress and exert positive effects in cancer and cardiovascular and chronic inflammatory diseases. Apigenin, kaempferol and resveratrol present in fruits, vegetables and grain were investigated for their effect on the synthesis of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) at transcriptional level in lipopolysaccharide (LPS)-stimulated J774.2 macrophages. Apigenin (30 microM), kaempferol (30 microM) and resveratrol (50 microM) significantly decreased the number of TNF-alpha mRNA copies in LPS-activated J774.2 macrophages. Apigenin and kaempferol caused inhibition of IL-1beta gene expression in J774.2 macrophages, but resveratrol was ineffective. These results indicate that apigenin, kaempferol and resveratrol exert inhibitory effects on the TNF-alpha and except for of resveratrol on IL-1beta gene expression in J774.2 macrophages at the transcriptional level. In addition, the studied compounds may be the mediators responsible for protective role of a diet high in fruits and vegetables in the cardiovascular and inflammatory diseases.

Effect of kaempferol on the production and gene expression of monocyte chemoattractant protein-1 in J774.2 macrophages.

Monocyte chemoattractant protein-1 (MCP-1) is produced by activated macrophages, and is involved in pathogenesis of cardiovascular and neurodegenerative disorders. There is a need to develop drugs that inhibit excessive infiltration of monocytes and lymphocytes to the arterial wall and central nervous system. The aim of this study was to evaluate the effect of kaempferol on the (MCP-1) gene expression and MCP-1 protein release by J774.2 macrophage cultures in vitro. Kaempferol given both before and after lipopolysaccharide (LPS) administration reduced secretion of MCP-1. Kaempferol administered before LPS stimulation significantly decreased the number of copies of MCP-1 mRNA. The results suggest that kaempferol inhibits MCP-1 production at the transcriptional level, and that this is an additional anti-inflammatory mechanism of action of this flavonoid.

Synthesis and tumoricidal activity evaluation of new morin and quercetin sulfonic derivatives.

Flavonoids are a group of naturally occurring compounds with interesting medical properties, such as antiinflammatory, antiallergic, antiviral, antibacterial and antitumor activities. In our experiments we were trying to examine the tumoricidal activity of newly synthesized derivatives of two flavonoids: 3,5,7,2',4'-pentahydroxyflavone (morin) and 3,5,7,3',4'-pentahydroxyflavone (quercetin). These derivatives were: natrium salt of morin-5'-sulfonic acid (NaMSA), natrium salt of quercetin-5'-sulfonic acid (NaQSA), complex of Mg2+ with quercetin-5'-sulfonic acid (QSA), complex of iron(II) with QSA. The antitumor activity of these agents was tested in vitro on two cell lines: L1210--murine lymphocytic leukaemia and P-815--murine mastocytoma. Our experiments showed that sulfonic derivatives of these two flavonoids were less potent than the original agents in their cytostatic and cytotoxic activities. However, their solubility in water was greater than that of the original agents and higher culture medium concentration of these derivatives was obtained. The results indicate that the ability of flavonoids to act tumoricidally is reciprocally correlated with their lipophilicity.