Crotonis Fructus and Its Constituent, Croton Oil, Stimulate Lipolysis in OP9 Adipocytes

Figure 2

Signaling pathway involved in the lipolytic effects of CFE and CO. Differentiated OP9 adipocytes were pretreated with 20 μM H89 and then treated with 20 μg/mL of CFE, 10 μg/mL of CO, and 200 nM ISO for 12 h. Glycerol release (black bar) quantification is the same as Figure 1 legends and real-time PCR for hormone sensitive lipase (HSL) mRNA (gray bar) was carried out using a specific primer for HSL as described in Materials and Methods. Protein expression levels for p-HSLS563 and HSL (lower panel) were subjected to western blot analysis by using specific antibodies. Data are expressed as the mean ± SD of four independent experiments and a percentage of vehicle (DMSO-)treated control cells (CON). ^*, ^** versus CON; ^## versus CFE-treated group; and versus CO-treated group.