Gastrodin Inhibits Expression of Inducible NO Synthase, Cyclooxygenase-2 and Proinflammatory Cytokines in Cultured LPS-Stimulated Microglia via MAPK Pathways
Figure 7
Inhibitory effects of gastrodin on the LPS-induced phosphorylation expression of IκB-α(A), CREB (B), ERK1/2 (C), JNK(D) and p38 MAPK (E) in BV-2 cells.
Approximately 1×106 cells/ml were seeded in six-well plates and incubated until 80% confluency. Cells were pre-treated with gastrodin (30, 40, and 60 µM) for 1 h, then exposed to 1 µg/ml LPS for 30 min. Cell lysates (50 µg protein) were prepared and subjected to Western blot analysis by using antibodies specific for phosphorylated forms of IκB-α, CREB, ERK1/2, JNK and p38 MAPK (shown as phospho-IκB-α, etc.) as described in the methods. Equivalent loading of cell lysates was determined by reprobing the blots with anti-β-actin, total ERK1/2, JNK or p38 MAPK antibodies. The relative protein levels were quantified by scanning densitometry and normalized to β-actin, total ERK1/2, JNK or p38 MAPK. The values shown are mean ± SEM of data from three independent experiments. # Significant compared with control alone, p<0.05. *Significant compared with LPS alone, p<0.05.