Journal: Oxidative Medicine and Cellular Longevity
Article Title: Pterostilbene Attenuates Cocultured BV-2 Microglial Inflammation-Mediated SH-SY5Y Neuronal Oxidative Injury via SIRT-1 Signalling
doi: 10.1155/2020/3986348
Figure Lengend Snippet: LPS induces BV-2 activation and SH-SY5Y injury in the coculture system, and PTE shows protective effects after LPS stimulation. SH-SY5Y cells were cocultured with BV-2 cells, LPS (100 ng/mL), or both BV-2 cells and LPS for 24 h, separately. (a) The morphology and viability of SH-SY5Y cells were observed under an inverted/phase-contrast microscope or using a CCK-8 assay, respectively. The BV-2 cells were preincubated with vehicle control or PTE (2.5, 5.0, or 10.0 μ M) for 2 h, followed by LPS stimulation and coculturing with SH-SY5Y cells for 24 h. The viability of (b) SH-SY5Y and (c) BV-2 cells was measured using a CCK-8 assay. (d) The LDH release in supernatants was determined using an LDH release assay and expressed as the percentage of maximum. (e) The apoptosis of SH-SY5Y cells was assessed using a TUNEL assay. The apoptotic nuclei were stained with TUNEL (green), and all nuclei were stained with DAPI (blue). (f) Apoptotic rate was computed as a percentage of the TUNEL- to the DAPI-positive nuclei. Data are shown as mean ± SEM. n = 6. α p < 0.05, compared with the control. β p < 0.05, compared with the a-BV-2. γ p < 0.05, compared with the PTE 2.5 μ M. LPS: lipopolysaccharide; a-BV-2: LPS-activated BV-2 coculture group.
Article Snippet: PTE, EX527, and DCFH-DA and DAPI fluorescent probes were obtained from Sigma-Aldrich (St. Louis, MO, USA).
Techniques: Activation Assay, Microscopy, CCK-8 Assay, Lactate Dehydrogenase Assay, TUNEL Assay, Staining