In vitro immunomodulatory activity, cytotoxicity and chemistry of some central European polypores

Context: Some mushrooms of the order Polyporales are known for their immunomodulatory actions. Objective: The objective of this study is to evaluate the in vitro phagocytic and cytotoxic effects of extracts from polyporales native to Central Europe. Materials and methods: The effects of ethanol extracts from 27 polypore species on opsonized zymosan-induced phagocytosis of isolated human neutrophils were tested by a chemiluminescence method. Colon epithelial cell lines, Caco-2 and HT-29, were used for cytotoxicity assays, and extracts were chemically characterized in terms of total phenolic and β-glucan content. Results: We observed phagocytosis or respiratory burst enhancing activity in 17 extracts, of which five species, namely Aurantiporus fissilis (Berk. & M.A. Curtis) H. Jahn ex Ryvarden, Trametes gibbosa (Pers.) Fr., Piptoporus betulinus (Bull.) P. Karst, Neolentinus lepideus (Fr.) Redhead & Ginns, Polyporus squamosus (Huds.) Fr., significantly increased phagocytosis in granulocytes by 205, 181, 158, 155 and 141%, respectively. The β-glucan content of the three most potent extracts was 58, 42 and 74 mg/g, respectively, and the polyphenol content was 155.6, 133.5 and 155.2 μmol of gallic acid equivalent/g, respectively. Some extracts showed cytotoxic activity, with higher cytotoxicity in Caco-2 than in HT-29 cells. Pycnoporus cinnabarinus (Jacq.) P. Karst. extract was cytotoxic to both cell lines, with IC50 values of 81 and 31 μg/mL, respectively. Discussion and conclusion: The most promising extracts were from N. lepideus and Polyporus squamosus, which are edible species and may be considered safe. Our findings support their use as culinary preparations or food supplements for various immunological gut disorders

Bersavin: Nový bisbenzylisochinolinový alkaloid s cytotoxickými, antiproliferativními a apoptózovými účinky na lidské leukemické buňky

Bersavine is the new bisbenzylisoquinoline alkaloid isolated from the Berberis vulgaris L. (Berberidaceae) plant. The results of cytotoxicity screening 48 h post-treatment showed that bersavine considerably inhibits the proliferation and viability of leukemic (Jurkat, MOLT-4), colon (HT-29), cervix (HeLa) and breast (MCF-7) cancer cells with IC50 values ranging from 8.1 to 11 mu M. The viability and proliferation of leukemic Jurkat and MOLT-4 cells were decreased after bersavine treatment in a time- and dose-dependent manner. Bersavine manifested concentration-dependent antiproliferative activity in human lung, breast, ovarian and hepatocellular carcinoma cell lines using a xCELLigence assay. Significantly higher percentages of MOLT-4 cells exposed to bersavine at 20 mu M for 24 h were arrested in the G1 phase of the cell cycle using the flow cytometry method. The higher percentage of apoptotic cells was measured after 24 h of bersavine treatment. The upregulation of p53 phosphorylated on Ser392 was detected during the progression of MOLT-4 cell apoptosis. Mechanistically, bersavine-induced apoptosis is an effect of increased activity of caspases, while reduced proliferation seems dependent on increased Chk1 Ser345 phosphorylation and decreased Rb Ser807/811 phosphorylation in human leukemic cells.Bersavin je nový bisbenzylisochinolinový alkaloid izolovaný z rostliny Berberis vulgaris L. (Berberidaceae). Výsledky cytotoxického screeningu po 48 hodinách ukázaly, že bersavin významně inhibuje proliferaci a životaschopnost leukemických (Jurkat, MOLT-4), tlustého střeva (HT-29), děložního čípku (HeLa) a karcinomu prsu (MCF-7) s hodnotami IC50 v rozmezí od 8,1 do 11 uM. Viabilita a proliferace leukemických buněk Jurkat a MOLT-4 byla po ovlivnění bersavinem snížena v závislosti na čase a dávce. Bersavin vykazoval na koncentraci závislou antiproliferativní aktivitu u lidských nádorových linií plic, prsu, vaječníků a hepatocelulárního karcinomu pomocí testu xCELLigence. Významně vyšší procento buněk MOLT-4 ovlivněných bersavinem v dávce 20 uM po dobu 24 hodin bylo zastaveno ve fázi G1 buněčného cyklu měřeného pomocí průtokové cytometrie. Vyšší procento apoptotických buněk bylo zjištěno za 24 hodin po ovlivnění bersavinem. Zvýšená exprese p53 fosforylovaného na Ser392 byla detekována během progrese apoptózy u buněk MOLT-4. Apoptóza indukovaná bersavinem je důsledkem zvýšené aktivity kaspáz, zatímco, snížená proliferace závisí pravděpodobně na zvýšené fosforylaci Chk1 Ser345 a snížené fosforylaci Rb Ser807 / 811 u lidských leukemických buněk

Porovnání cytotoxicity chelidoninu a homochelidoninu, dimethoxy analoga izolovaného z Chelidonium majus L. (Papaveraceae), vůči lidským leukemickým a plicním nádorovým buňkám

In this study the effects of naturally occurring homochelidonine in comparison to chelidonine on cell cycle progression and cell death in leukemic T-cells with different p53 status are described. We found that homochelidonine and chelidonine displayed significant cytotoxicity in examined blood cancer cells with the exception of HEL 92.1.7 and U-937 exposed to homochelidonine. Unexpectedly, homochelidonine and chelidonine-induced cytotoxicity was more pronounced in Jurkat cells contrary to MOLT-4 cells. Homochelidonine showed an antiproliferative effect on A549 cells but it was less effective compared to chelidonine. Biphasic dose-depended G1 and G2/M cell cycle arrest along with the population of sub-G1 was found after treatment with homochelidonine in MOLT-4 cells. In variance thereto, an increase in G2/M cells was detected after treatment with homochelidonine in Jurkat cells. Treatment with chelidonine induced cell cycle arrest in the G2/M cell cycle in both MOLT-4 and Jurkat cells. MOLT-4 and Jurkat cells treated with homochelidonine and chelidonine showed features of apoptosis such as phosphatidylserine exposure, a loss of mitochondrial membrane potential and an increase in the caspases -3/7, -8 and -9. Western blots indicate that homochelidonine and chelidonine exposure activates Chk1 and Chk2. Studies conducted with fluorescence microscopy demonstrated that chelidonine and homochelidonine inhibit tubulin polymerization in A549 cells. Collectively, the data indicate that chelidonine and homochelidonine are potent inducers of cell death in cancer cell lines, highlighting their potential relevance in leukemic cells.Tato práce porovnává cytotoxicitu chelidoninu a homochelidoninu vůči lidským leukemickým a plicním nádorovým buňkám. Výsledky studie naznačují, že chelidonin a homochelidonin jsou účinnými aktivátory apoptózy u nádorových buněčných linií

In vitro a in silico stanovení inhibiční aktivity thaliktrikavinu a kanadinu vůči acetylcholinesteráze a odhad jejich přestupu přes hematoencefalickou bariéru

In recent studies, several alkaloids acting as cholinesterase inhibitors were isolated from Corydalis cava (Papaveraceae). Inhibitory activities of (+)-thalictricavine (1) and (+)-canadine (2) on human acetylcholinesterase (hAChE) and butyrylcholinesterase (hBChE) were evaluated with the Ellman's spectrophotometric method. Molecular modeling was used to inspect the binding mode of compounds into the active site pocket of hAChE. The possible permeability of 1 and 2 through the blood-brain barrier (BBB) was predicted by the parallel artificial permeation assay (PAMPA) and logBB calculation. In vitro, 1 and 2 were found to be selective hAChE inhibitors with IC50 values of 0.38 +/- 0.05 mu M and 0.70 +/- 0.07 mu M, respectively, but against hBChE were considered inactive (IC50 values > 100 mu M). Furthermore, both alkaloids demonstrated a competitive-type pattern of hAChE inhibition and bind, most probably, in the same AChE sub-site as its substrate. In silico docking experiments allowed us to confirm their binding poses into the active center of hAChE. Based on the PAMPA and logBB calculation, 2 is potentially centrally active, but for 1 BBB crossing is limited. In conclusion, 1 and 2 appear as potential lead compounds for the treatment of Alzheimer's disease.Z Corydalis cava (Papaveraceae) bylo izolováno několik alkaloidů půsbících jako inhibitory cholinesterázy. Inhibiční aktivity (+) - thaliktrikavinu (1) a (+) - kanadinu (2) na lidskou acetylcholinesterázu (hAChE) a butyrylcholinesterázu (hBChE) byly hodnoceny pomocí Ellmanovy spektrofotometrické metody. K ověření způsobu vazby testovaných sloučenin do aktivního místa hAChE bylo použito molekulové modelování. Možná permeabilita 1 a 2 hematoencefalickou bariérou (HEB) ​​byla předpovězena pomocí testu paralelní umělé permeace (PAMPA) a logBB. In vitro bylo zjištěno, že 1 a 2 jsou selektivní inhibitory hAChE s hodnotami IC50 0,38 +/- 0,05 uM, respektive 0,70 +/- 0,07 uM, ale při inhibici hBChE byly považovány za neaktivní (hodnoty IC50> 100 uM). Oba alkaloidy prokázaly kompetitivní typ inhibice hAChE a vážou se s největší pravděpodobností na stejném AChE sub-místě jako jeho substrát. In silico dokovací experimenty potvrdily vazebné pozice v aktivním místě AChE. Na základě výpočtu PAMPA a logBB je 2 potenciálně centrálně aktivní, ale pro 1 je přechod HEB omezen