Neuroprotective Effects ofCastanea sativaMill. Bark Extract in Human Neuroblastoma Cells Subjected to Oxidative Stress

Abstract

One of the major features of neurodegenerative disease is the selective vulnerability of different neuronal populations that are affected in a progressive and often stereotyped manner. Despite the susceptible neuronal population varies between diseases, oxidative stress is implicated as the major pathogenic process in all of them. Natural Extract of Castanea sativa Mill. bark (ENC), recently characterized in its phenolic composition, acts as antioxidant and cardioprotective agent. Its neuroprotettive properties, however, have never been investigated. The aim of this study was to assess neuroprotection of ENC in in vitro models of oxidative-stress-mediate injury. Human neuroblastoma SH-SY5Y cells treated with glutamate (50 mM for 24 h) or hydrogen peroxide (25 μM for 1 h followed by 24 with medium) were used. The results showed that the addition of ENC (1-50 μg/ml) to cell medium before the neuronal damage provided neuroprotection in both experimental models used, while its addition after the injury was ineffective. In conclusion, the present results suggest that ENC could be a valuable support as dietary supplement, combining beneficial preventive neuroprotettive effects with a high antioxidant activity.

Castanea sativa Mill. bark extract exhibits chemopreventive properties triggering extrinsic apoptotic pathway in Jurkat cells

“…Even though CSM wood, bark and leaf extracts have recently attracted interest due to their photoprotective, neuroprotective, cardioprotective and antioxidant properties that suggest their suitability for the prevention of chronic and degenerative diseases [22, 23, 41], to our knowledge no study has previously evaluated the pro-apoptotic effect of CSM extracts on cancer cells. In this study, we evaluated the effects of CSM bark extract in human T leukemia cells.…”

Apoptotic-Induced Effects of Acacia Catechu Willd. Extract in Human Colon Cancer Cells

“…In order to monitor these changes caused by AC, HT-29 cells were examined by using a phase-contrast light microscope, and the results were evaluated using the grade scale described in USP 28 (United States Pharmacopeia edition 2005) (grades 0-4) for the assessment of the cytotoxic potentials of the materials as follows: grade 0-no reactivity (discrete intracytoplasmic granules, no cell lysis); grade 1-slight reactivity (no more than 20% of the cells are round, loosely attached and without intracytoplasmic granules; occasional lysed cells are present); grade 2-mild reactivity (no more than 50% of the cells are round and devoid of intracytoplasmic granules; no extensive cell lysis and empty areas between cells); grade 3-moderate (up to 70% of cells are rounded or lysed); grade 4-severe (nearly complete destruction of the cells). [62,63].…”

“…Flow cytometry techniques, such as cell cycle and sub-G0/G1 population-analysis, and annexin V/propidium iodide (AV/PI) and 4 ,6-diamidino-2-phenylindole (DAPI) staining, were applied to identify apoptotic cells. HT-29 cells (5 × 10 5 cells/well, final volume of 2 mL) were treated with AC for 24 h, and then the analysis was performed by using protocols previously described [62]. Alexa fluor 488™-AV/PI double staining kit (Life Tecnologies Italia, Monza, Italy) was used to detect the externalization of phosphatidylserine in apoptotic cells [64].…”

“…Viable cells were both AV and PI-negative; cells in early apoptosis were AV-positive and PI-negative; cells in late apoptosis were both AV and PI-positive; necrotic cells were PI-positive and AV-negative. Apoptosis was assessed also by analysing changes in nuclear morphology with the DAPI staining kit (Life Tecnologies Italia, Monza, Italy), as previously described [62].…”

Vascular Toxicity Risk Assessment of MC18 and MC70, Novel Potential Diagnostic Tools for In Vivo PET Studies

“…Therefore, A7r5 or EA.hy926 cells were examined by a phase‐contrast light microscope and photographed with a digital camera 24, 48 or 72 hr after treatment. To assess the cytotoxic potential of compounds, images were evaluated using the grade scale described in the United States of America Pharmacopeia 28, edition 2005, as follows: grade 0 – none reactivity (discrete intracytoplasmic granules, no cell lysis); grade 1 – slight reactivity (<20% of the cells are round, loosely attached and without intracytoplasmic granules; occasional lysed cells are present); grade 2 – mild reactivity (less than 50% of the cells are round and devoid of intracytoplasmic granules, no extensive cell lysis and empty areas between cells); grade 3 – moderate reactivity (up to 70% of cells are rounded or lysed); and grade 4 – severe reactivity (nearly complete destruction of cells) .…”