A Chinese Herbal Preparation, Xiao-Er-An-Shen Decoction, Exerts Neuron Protection by Modulation of Differentiation and Antioxidant Activity in Cultured PC12 Cells

Figure 4

XEASD induces CREB phosphorylation on PC12 cells. (a) Cultured PC12 cells were serum starved over 5 hours before the treatment with XEASD extract (1 mg/mL) or NGF (50 ng/mL) for different time. Total CREB and phosphorylated CREB (both at ~ 43 kDa) were determined by using specific antibodies. (b) Cultured PC12 cells with serum starvation for over 5 hours were pretreated with or without H89 (5 μM; a PKA inhibitor) for 3 hours prior to the treatment with NGF (50 ng/mL) and XEASD (1.0 mg/mL) for 10 minutes. Total CREB and phosphorylated CREB were determined by using specific antibodies (upper panel). Quantification plot was indicated in histograms (lower panel). Values are expressed as the fold of change (x Basal) against the control (no treatment; set as 1). Mean ± SEM; n = 5. Statistical comparison was made with the H89-treated group; < 0.01.

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(b)