Protective Effect of Arzanol against H2O2-Induced Oxidative Stress Damage in Differentiated and Undifferentiated SH-SY5Y Cells

Oxidative stress can damage neuronal cells, greatly contributing to neurodegenerative diseases (NDs). In this study, the protective activity of arzanol, a natural prenylated α-pyrone-phloroglucinol heterodimer, was evaluated against the H2O2-induced oxidative damage in trans-retinoic acid-differentiated (neuron-like) human SH-SY5Y cells, widely used as a neuronal cell model of neurological disorders. The pre-incubation (for 2 and 24 h) with arzanol (5, 10, and 25 μM) significantly preserved differentiated SH-SY5Y cells from cytotoxicity (MTT assay) and morphological changes induced by 0.25 and 0.5 mM H2O2. Arzanol reduced the generation of reactive oxygen species (ROS) induced by 2 h oxidation with H2O2 0.5 mM, established by 2′,7′-dichlorodihydrofluorescein diacetate assay. The 2 h incubation of differentiated SH-SY5Y cells with H2O2 determined a significant increase in the number of apoptotic cells versus control cells, evaluated by propidium iodide fluorescence assay (red fluorescence) and NucView® 488 assay (green fluorescence). Arzanol pre-treatment (2 h) exerted a noteworthy significant protective effect against apoptosis. In addition, arzanol was tested, for comparison, in undifferentiated SH-SY5Y cells for cytotoxicity and its ability to protect against H2O2-induced oxidative stress. Furthermore, the PubChem database and freely accessible web tools SwissADME and pkCSM-pharmacokinetics were used to assess the physicochemical and pharmacokinetic properties of arzanol. Our results qualify arzanol as an antioxidant agent with potential neuroprotective effects against neuronal oxidative stress implicated in NDs

Fatty Acid Ratios as Parameters to Discriminate Between Normal and Tumoral Cells and Compare Drug Treatments in Cancer Cells

The fatty acid (FA) composition of cell membranes represents a metabolic biomarker. However, the FA profile reproducibility in cell cultures remains a significant challenge. In this study, cell FA ratios are validated as metabolic markers alternative to cell FA. To this goal, cell samples belonging to cancer HeLa cells and normal 3T3 fibroblasts, from various experimental sets, are analyzed by a high-performance liquid chromatography system coupled with a photodiode array detector and evaporative light scattering detector (HPLC-DAD/ELSD), and the ratios among the main FA are calculated. Principal component analysis (PCA) separately performed on FA and FA ratio data indicates similar clustering of cell samples concerning the cell type. Moreover, similar scores values t[1] and t[2] and graphical distances are calculated in the PCA plots separately performed on FA and FA ratios measured in cancer HeLa cells subjected to various antitumoral compounds. Last, PCA applied to selected FA ratios measured in various cell lines, obtained in similar experimental conditions, allows to discriminate between normal and tumoral cells. The results substantiate FA ratios as a cell-specific fingerprint, characterized by reproducibility across intra-laboratory conditions, useful for cell characterization, discrimination between normal and tumoral cells, and the comparison of different drug treatments. Practical Applications: The reproducibility of the fatty acid (FA) profile in cell cultures remains a significant challenge. Results obtained from this study improve knowledge about the role of the FA ratio profile as a cell-specific fingerprint characterized by reproducibility across intra-laboratory conditions. The characterization of the specific FA ratio profile of a cell culture, under standardized experimental conditions, can facilitate the comparative evaluation of cell data sets for nutritional, metabolic, and pharmacological studies, overcoming differences in cell culture conditions and FA extraction/analytical procedures

Targeting phosphoglycerate kinases by tatridin A, a natural sesquiterpenoid endowed with anti-cancer activity, using a proteomic platform

Tatridin A (TatA) is a germacrane sesquiterpenoid containing one E-double bond and one Z-double bond in its 10-membered ring, which is fused to a 3-methylene-dihydrofuran-2-one moiety. Tatridin A bioactivity has been poorly investigated despite its interesting chemical structure. Here, a functional proteomic platform was adapted to disclose its most reliable targets in leukemia monocytic cells, and phosphoglycerate kinases were recognized as the most affine enzymes. Through a combination of limited proteolysis and molecular docking, it has been discovered that tatridin A interacts with the active domains of phosphoglycerate kinase 1, altering its hinge region, and it can be accountable for tatridin A inhibition potency on enzyme activity. A more detailed tatridin A biological profile showed that it is also fully active against gastric cancer cells, downregulating the mRNA levels of chemokine receptor 4 and β-catenin and inhibiting the invasiveness of living KATO III cells as a direct consequence of phosphoglycerate kinase 1 antagonism

Mapping the expression of transient receptor potential channels across murine placental development.

Funder: Onderzoeksraad, KU Leuven; doi: http://dx.doi.org/10.13039/501100004497Funder: Centre of Trophoblast Research, University of CambridgeTransient receptor potential (TRP) channels play prominent roles in ion homeostasis by their ability to control cation influx. Mouse placentation is governed by the processes of trophoblast proliferation, invasion, differentiation, and fusion, all of which require calcium signaling. Although certain TRP channels have been shown to contribute to maternal-fetal transport of magnesium and calcium, a role for TRP channels in specific trophoblast functions has been disregarded. Using qRT-PCR and in situ hybridisation, the spatio-temporal expression pattern of TRP channels in the mouse placenta across gestation (E10.5-E18.5) was assessed. Prominent expression was observed for Trpv2, Trpm6, and Trpm7. Calcium microfluorimetry in primary trophoblast cells isolated at E14.5 of gestation further revealed the functional activity of TRPV2 and TRPM7. Finally, comparing TRP channels expression in mouse trophoblast stem cells (mTSCs) and mouse embryonic stem cells (mESC) confirmed the specific expression of TRPV2 during placental development. Moreover, TRP channel expression was similar in mTSCs compared to primary trophoblasts and validate mTSC as a model to study TRP channels in placental development. Collectivity, our results identify a specific spatio-temporal TRP channel expression pattern in trophoblasts, suggesting a possible involvement in regulating the process of placentation