Investigation of Pentylenetetrazol (PTZ)-induced toxicity in primary cortical neurons. (A) Immunofluorescence detection of primary neuronal cells and bright field of the same area. Cells were stained with mouse anti-MAP2 antibody (green) and rabbit anti-GFAP antibody (red, detection of astrocytes). The nuclei are stained with DAPI (blue). (B) Observation of morphological changes of neurons using a contrast microscope at 7 days in vitro (DIV). The cells were treated with 0, 7.5, 15, 30, and 60 mM PTZ for 24 h at 7 DIV. (C) PTZ-induced cellular toxicity in neurons. Cell viability is presented as a percentage of control, and each value represents the mean ± SD of three independent experiments. Bar: 100 μm.

Otophylloside N attenuated PTZ-induced cell injury on primary cortical neurons. The neurons were treated with indicated concentrations of PTZ or PTZ + Otophylloside N (OtoN) for 24 h at 7 DIV. (A) Immunofluorescence detection of morphological changes of neurons and bright field of the same area. The cells were immunostained with neuronal anti-β-Tubulin marker (green) and nuclear DAPI tag (blue). (B) OtoN inhibited PTZ-induced toxicity in neurons. Cell viability is presented as a percentage of the control, and each value represents the mean ± SD of three independent experiments. (C) OtoN decreased PTZ-induced LDH intracellular efflux in neurons. The LDH efflux level is presented as a percentage of the control, and each value represents the mean ± SD of three independent experiments. ^##P < 0.01 compared with the control. ^∗∗P < 0.01 compared with the 30 mM PTZ treatment. Bar: 100 μm.

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