The concentration-dependent effect of anethole on collagen, MMP-2 and GAG in human skin fibroblast cultures.

PURPOSE: In aging skin and some skin disorders, components of skin extracellular matrix (ECM) are disturbed and therefore research to find skin drugs is important. Evaluation of anethole impact on collagen, GAGs and MMP-2 in human skin fibroblasts was the aim of this study. MATERIALS AND METHODS: For collagen assay the Sircol dye, 5-[3H]proline and real time-PCR were used. MMP-2 activity was detected by zymography. GAG concentration was determined using 1,9-dimethylmethylene blue (DMMB). Cell viability was assayed with MTT. RESULTS: In cells treated with 1 and 10 µM anethole, a significant increase in collagen synthesis was demonstrated. In contrast, collagen synthesis was significantly decreased in cells exposed to 100 µM anethole. Similar alterations were found in collagen type I expression. The concentration of collagen secreted into the medium was higher only in cells exposed to 1 µM anethole, while it was lower under the influence of higher compound concentrations. It may be due to the lack of pro-MMP-2 activation at 1 µM and a significant increase in the level of MMP-2 at 10 and 100 µM anethole. GAG concentration was reduced under the influence of 100 µM anethole, whereas anethole at lower concentrations revealed the ability to prevent H2O2-induced GAG increase. No significant cytotoxicity of anethole to fibroblasts was noted. CONCLUSIONS: Our findings demonstrate the concentration-dependent action of anethole on the crucial components of ECM in cultured skin fibroblasts, which may be somewhat beneficial and may possibly be developed towards a therapeutic use in some skin disorders.

Lotus tetragonolobus and Maackia amurensis lectins influence phospho-IκBα, IL-8, Lewis b and H type 1 glycoforms levels in H. pylori infected CRL-1739 gastric cancer cells.

PURPOSE: Attachment of Helicobacter pylori to the mucous epithelial cells and the mucous layer is said to be a crucial step for infection development. Sugar antigens of gastric mucins (MUC5AC, MUC1) can act as receptors for bacterial adhesins. The aim of the study was to investigate if Lotus tetragonolobus and Maackia amurensis lectins influence the level of MUC1, MUC5AC, Lewis b, H type 1, sialyl Lewis x, phospho-IκBα and interleukin 8 in Helicobacter pylori infected gastric cancer cells. MATERIALS AND METHODS: The study was performed with one clinical H. pylori strain and CRL-1739 gastric cancer cells. To assess the levels of mentioned factors immunosorbent ELISA assays were used. RESULTS: Coculture of cells with bacteria had no clear effect on almost all examined structures. After coculture with H. pylori and lectins, a decrease of the level of both mucins, Lewis b and H type 1 antigens was observed. Lectins addition had no effect on sialyl Lewis x. Maackia amurensis caused slight increase of phospho-IκBα while interleukin 8 level was decreased. CONCLUSIONS: Lotus tetragonolobus and Maackia amurensis lectins can mediate in binding of Helicobacter pylori to gastric epithelium.

Expression of MUC1 mucin in human umbilical vein endothelial cells (HUVEC).

Mucin 1 (MUC1) is a membrane-bound glycoprotein that is expressed by various epithelial cell types. MUC1 functions include modulation of cell adhesion, signal transduction, lubrication and hydration of epithelial surfaces, and their protection from infection. In this study we demonstrated that MUC1 is expressed in human umbilical vein endothelial cells (HUVECs) and could be released/shed from cellular membrane. MUC1 presence in these cells was verified using three methods: Western blotting, flow cytometry and metabolic labeling. We also showed that mucin expression is stimulated by proinflammatory cytokines: about a 2-fold increase was observed after TNF-α treatment and lower after IFN-γ alone and in combination with TNF-α treatment. It can be assumed that the presence of MUC1 in endothelial cells may have an important role in the interactions with different cell types in physiological and pathological processes.

Characterisation of glycoforms of ascitic fluids in benign and malignant diseases.

OBJECTIVES: To evaluate the differences in glycoforms of ascitic fluids which can be considered as useful in discrimination between benign and malignant patients. DESIGN AND METHODS: The investigations were carried out on 17 benign and 13 malignant patients. To obtain high molecular weight material, gel exclusion chromatography was applied. To evaluate the relative amounts of different glycoforms, ELISA tests with biotinylated lectins were used. RESULTS: The fucosylation pattern was found to be similar in malignant and benign group. Fucose linked by alpha 1-6 bond was the most abundant structure. Sialylation was found to be more typical for malignant patients. Alpha 2-6 bond was observed on higher level than alpha 2-3 linkage. T and Tn antigens were present on rather low level with a slight prevalence of T antigen in malignant group. The glycoforms recognized by DSA lectin were more numerous in benign patients. CONCLUSIONS: The evaluation of the level of some ascitic fluids glycoforms can be useful in differentiation between malignant and benign diseases.

The effect of Helicobacter pylori eradication treatment on the MUC 1 and Lewis antigens level in human gastric juice: a preliminary study.

Helicobacter pylori is considered as a causative agent of gastritis, duodenal and gastric ulcers, and gastric cancer. During inflammation, association of the pathogen of gastric epithelial cells and mucins is considered important. It was postulated that Lewis b structures of secretory MUC 5AC mucin can be a receptor for the bacterium. Some authors also suggest that epithelial MUC 1 mucin may be implicated in the mechanism of infection. The main aim of our work was to support this last suggestion by evaluation of the possible changes in MUC 1 and Lewis a and b levels in gastric juice before and at the end of eradication treatment. The gastric juices of ten examined patients were chromatographed on a Sepharose 4 B column, electrotransferred on Immobilon P membranes, and assessed for MUC 1 and Lewis a and b structures using monoclonal antibodies. In 90% of examined patients, higher amounts of MUC 1 mucin were observed at the end of eradication treatment. Similar results for Lewis a and b structures were found. In the case of MUC 1 and Lewis b, the differences were statistically significant. Helicobacter pylori influences expression of the soluble form of MUC 1 mucin and Lewis a and b structures present in gastric juice.