The role of airway wall remodelling in bronchial asthma is well established. Myofibroblasts, the cells displaying features intermediate between fibroblasts and smooth muscle cells, are involved in this process but the mechanism of myofibroblasts activation in the onset of the disease remains obscure. Myofibroblasts can differentiate from various cell types, including resident fibroblasts, and the fibroblasts to myofibroblasts transition (FMT) can be reproduced in vitro. We aimed to investigate the process of FMT in human bronchial fibroblasts (HBF) derived from non-asthmatic (n = 7) and asthmatic (n = 7) subjects. We also tested whether cell-cell contacts affect FMT by using N-cadherin blocking antibody. HBF plated in low or high cell density were treated with TGF-β(1) up to one week to induce FMT. The percentage of myofibroblsts was counted and expression of α-smooth muscle actin was evaluated by cytoimmunofluorescence, flow cytometry and immunobloting. We demonstrated that the intensity of FMT induced by TGF-β(1)in vitro was strongly enhanced in asthmatic as compared to non-asthmatic HBF populations. This process was facilitated by low cell plating density in both groups of cultures. Furthermore, we proved that neither HBF-conditioned medium nor growth arrest in G(0)/G(1) phase of cell cycle could stop the TGF-β(1)-induced FMT in asthmatic cell populations. However, even in sparse asthmatic HBF, the blocking of N-cadherin resulted in the inhibition of FMT. Our findings show for the first time that the initial absence or an induced loss of cell-cell adhesions in asthmatic HBF populations is important for the completion of FMT.
Lentinula edodes (shiitake), an edible and medicinal mushroom, was chosen for this study with the aim of evaluating the possibility of release of bioelements into artificial digestive juices and analyzing the anti-inflammatory properties. The extracts were prepared from fruiting bodies and biomass enriched with copper (Cu), zinc (Zn), and selenium (Se). The content of bioelements was analyzed by total reflection X-ray fluorescence method. Relatively low content of elements was observed in the fruiting bodies: Cu-1.6, Zn-7.6, and Se-0.12 mg/100 g d.w. compared to mycelial cultures. The anti-inflammatory properties were evaluated in RAW 264.7 cells. Based on the levels of cyclooxygenase 2 protein, nuclear factor erythroid 2-related factor 2, and peroxisome proliferator-activated receptor γ determined using Western blot technique, it was found that the addition of bioelements enhanced the anti-inflammatory properties of mycelium. This indicates that L. edodes cultured on a suitable medium may be used as a potential component of anti-inflammatory products. Keywords Shiitake. Biologically active elements. Extraction in digestive juices. Medicinal properties of mushrooms. RAW 264.7 cells * Bożena Muszyńska
Pleurotus eryngii (DC:Fr.) Quel. is a cultivated mushroom of high culinary value and medicinal properties. Mycelium of P. eryngii is characterized by the ability of effective bio-elements absorption from growth media so it could be biofortified with trace elements with a functional activity in the human body. In this study, the ability of P. eryngii mycelia from in vitro cultures as well as fruiting bodies were investigated in terms of their effectiveness in zinc and selenium accumulation. The effect of Se and Zn biofortification on productivity, chemical compounds, and bio-elements content of P. eryngii was determined as well. To enhance Se and Zn content in P. eryngii fruiting bodies and mycelia, substrates were supplemented with sodium selenite, at a concentration of 50 mg L−1, zinc sulfate, and zinc hydro-aspartate at a concentration of 87.2 and 100.0 mg L−1, respectively. Mentioned Zn concentrations contained the same amount of zinc(II) ions, namely 20 mg L−1. The content of organic compounds include phenolic compounds and lovastatin, which were determined by a high-performance liquid chromatography with diode-array detector (HPLC-DAD) and reverse phase high-performance liquid chromatography (RP-HPLC) method with UV detection. The ability of P. eryngii to accumulate zinc and selenium from the culture medium was demonstrated. The degree of accumulation of zinc turned out to be different depending on the type of salt used. The present study also showed that conducting mycelium of P. eryngii in in vitro culture, with a higher content of zinc ions, can result in obtaining the materials with better antioxidant ability. The results of this study can be used to develop the composition of growing media, which ensures the production of biomass with the desired composition of elements.
Medicinal mushrooms of the order Polyporales have a long history of use, which is evidenced by the finding of dissected fruiting bodies with Ötzi, who lived over 5000 years ago. Because of its valuable biological properties and its use in 18th and 19th‐century pharmacy, Fomitopsis officinalis used to be mass‐collected. Moreover, the large demand for larch wood and non‐wood materials (resin) caused an excessive exploitation of larch forests, which directly contributed to the disappearance of F. officinalis from its natural environment. The qualities of medicinal preparations obtained from the F. officinalis fruiting bodies are determined by the unique composition of its bioactive compounds, such as: triterpenoids, polysaccharides, organic acids, coumarins and phenolic compounds. It has been proved that both crude extracts and the compounds isolated from F. officinalis have a wide spectrum of therapeutic effects, including anti‐inflammatory, cytotoxic, and antimicrobial effects.
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