Journal: Molecules

Article Title: In Vitro Neuroprotective and Anti-Inflammatory Activities of Natural and Semi-Synthetic Spirosteroid Analogues

doi: 10.3390/molecules21080992

Figure Lengend Snippet: Anti-excitotoxic activity of compounds in glutamate-induced cell death. ( A ) PC12 cell cultures were exposed to an established excitotoxic concentration of glutamate (Glu) (20 mM), and some were also treated with each compound ( S9 and S15 semi-synthetic derivatives; E3, estriol; SL, solasodine; E1, estrone; T, testosterone; TDCA, taurodeoxycholic acid) or solvent control (DMSO). After 24 h of exposition to the damage and treatment, cellular viability was determined by the MTT assay; ( B ) Dose-response of Compound S15 in the same model of glutamate-induced excitotoxicity. Data are represented as the percent with respect to the untreated cells, representing the highest cell viability. * p < 0.05, ** p < 0.01 and *** p < 0.001, according to one-way ANOVA and Tukey’s post-hoc test.

Article Snippet: The amount of IL-1β released into the culture medium was measured using a sandwich ELISA system with antibodies specifics for rat IL-1β (Polyclonal Rat IL-1β/IL-1F2 Catalog Number AF-501-NA and Polyclonal Rat IL-1β/IL-1F2 Biotinylated Catalog Number BAF501, R&D Systems, Minneapolis, MN, USA), following the manufacturer’s instructions.

Techniques: Activity Assay, Concentration Assay, Solvent, Control, MTT Assay

Journal: Molecules

Article Title: In Vitro Neuroprotective and Anti-Inflammatory Activities of Natural and Semi-Synthetic Spirosteroid Analogues

doi: 10.3390/molecules21080992

Figure Lengend Snippet: Neuroprotective activity of compounds against cell damage induced by chemical hypoxia and glucose deprivation. PC12 cells were cultured in oxygen and glucose deprivation conditions (OGD), specifically in the absence of glucose and in the presence of 600 µM of potassium cyanide (KCN) for 1 h, and then, normal culture medium was reperfused to the cells. After 4 h of the reperfusion start, some PC12 cells were treated with solvent control (DMSO), solasodine (SL) or different concentrations of the S15 derivative. After 20 h of exposition to the reperfusion damage and treatment, cellular viability was determined by the MTT assay. Data are represented as the percent with respect to the untreated cells, representing the highest cell viability. Asterisks indicate statistical differences between the treatment and respective control condition (*** p < 0.001) according to one-way ANOVA and Tukey’s post-hoc test.

Article Snippet: The amount of IL-1β released into the culture medium was measured using a sandwich ELISA system with antibodies specifics for rat IL-1β (Polyclonal Rat IL-1β/IL-1F2 Catalog Number AF-501-NA and Polyclonal Rat IL-1β/IL-1F2 Biotinylated Catalog Number BAF501, R&D Systems, Minneapolis, MN, USA), following the manufacturer’s instructions.

Techniques: Activity Assay, Cell Culture, Solvent, Control, MTT Assay

Journal: Molecules

Article Title: In Vitro Neuroprotective and Anti-Inflammatory Activities of Natural and Semi-Synthetic Spirosteroid Analogues

doi: 10.3390/molecules21080992

Figure Lengend Snippet: In vitro anti-inflammatory activity of the S15 derivative in activated primary microglia. Microglia-enriched primary cultures were obtained from brains of adult male rats. Different concentrations of the S15 derivative, solvent control (DMSO) or 10 µM of dexamethasone (DX) as the positive control of inhibition were applied to some microglia cultures, 30 min before the activation of microglia cells with lipopolysaccharide (LPS, 1 µg/mL) and murine recombinant interferon gamma (IFNγ, 20 U/mL). Untreated (UT) microglia cells were included as the negative control of inhibition. After 24 h of exposition or not to the stimulus and treatment, the IL-1β level was quantified by ELISA ( A ); and the content of nitrite was determined by the Griess assay ( B ). Asterisks indicate statistical differences between the treatment and respective control condition (* p < 0.05, ** p < 0.01 and *** p < 0.001) according to one-way ANOVA and Tukey’s post-hoc test.

Article Snippet: The amount of IL-1β released into the culture medium was measured using a sandwich ELISA system with antibodies specifics for rat IL-1β (Polyclonal Rat IL-1β/IL-1F2 Catalog Number AF-501-NA and Polyclonal Rat IL-1β/IL-1F2 Biotinylated Catalog Number BAF501, R&D Systems, Minneapolis, MN, USA), following the manufacturer’s instructions.

Techniques: In Vitro, Activity Assay, Solvent, Control, Positive Control, Inhibition, Activation Assay, Recombinant, Negative Control, Enzyme-linked Immunosorbent Assay, Griess Assay