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Objective To investigate the role of extracellular signal-regulated kinase1/2(ERK1/2) pathway in the regulation of aquaporin 4(AQP4) expression in cultured astrocytes after scratch-injury. Methods The scratch-injury model was produced in cultured astrocytes of rat by a 10-μL plastic pipette tip. The morphological changes of astrocytes and lactate dehydrogenase(LDH) leakages were observed to assess the degree of scratch-injury. AQP4 expression was detected by immunofluorescence staining and Western blot, and phosphorylated-ERK1/2(p-ERK1/2) expression was determined by Western blot. To explore the effect of ERK1/2 pathway on AQP4 expression in scratch-injured astrocytes, 10 μmol/L U0126(ERK1/2 inhibitor) was incubated in the medium at 30 min before the scratch-injury in some groups. Results Increases in LDH leakage were observed at 1, 12, and 24 h after scratch-injury, and AQP4 expression was reduced simultaneously. Decrease in AQP4 expression was associated with a significant increase in ERK1/2 activation. Furthermore, pretreatment with U0126 blocked both ERK1/2 activation and decrease in AQP4 expression induced by scratch-injury. Conclusion These results indicate that ERK1/2 pathway down-regulates AQP4 expression in scratch-injured astrocytes, and ERK1/2 pathway might be a novel therapeutic target in reversing the effects of astrocytes that contribute to traumatic brain edema.
Methods To test the role of extracellular signal-regulated kinase 1/2 (ERK1 / 2) pathway in the regulation of aquaporin 4 (AQP4) expression in cultured astrocytes after scratch-injury. Methods The scratch-injury model was produced in cultured astrocytes of rat by a 10-μL plastic pipette tip. The morphological changes of astrocytes and lactate dehydrogenase (LDH) leakages were observed to assess the degree of scratch-injury. AQP4 expression was detected by immunofluorescence staining and Western blot, and phosphorylated-ERK1 / 2 To explore the effect of ERK1 / 2 pathway on AQP4 expression in scratch-injured astrocytes, 10 μmol / L U0126 (ERK1 / 2 inhibitor) was incubated in the medium at 30 min before the scratch-injury in some groups. Results Increases in LDH leakage were observed at 1, 12, and 24 h after scratch-injury, and AQP4 expression was reduced simultaneously. Decrease in AQP4 expression was associated with a significant increase in ERK1 / 2 activation. Furthermore, pretreatment with U0126 blocked both ERK1 / 2 activation and decrease in AQP4 expression induced by scratch-injury. Conclusion These results indicate that ERK1 / 2 pathway down-regulates AQP4 expression in scratch- injured astrocytes, and ERK1 / 2 pathway might be a novel therapeutic target in reversing the effects of astrocytes that contribute to traumatic brain edema.