Astrocyte Stretch Injury Model and Interventions

The protocol of astrocyte stretch injury model was similar to those described previously [22 (link)]. Equiaxial stretch (20% strain, 1.0 Hz frequency) was applied to cultured astrocytes for 1 h, by a Flexcell^® FX-5000™ Tension System (Flexcell, USA) with specific designed 6-well plates (BioFLEX^®). The culture media of astrocytes in stretch injury group and control group were collected at this time and will be used as conditioned medium. Again, to clarify the effects and signaling relationship of S100B, RAGE, ADAM17 on stretch injury, ONO-2506 (100 µM) [18 (link)], FPS-ZM1 (100 nM) [23 (link)] or TAPI-1 (50 µM) [24 (link)] were, respectively, applied for 6 h after stretch injury. To further verify the effect of S100B on astrocyte injury, exogenous S100B (1 µM) [8 (link)] was directly administrated to normal astrocytes for 6 h. The cell viability and relevant parameters were detected as indicated.