Lippia Sidoides stimulates mitochondrial biogenesis by elevating cAMP levels

In our discovery program we have the capability of screening botanicals for various activities that are related to anti-aging functionality. Within this program we have assays that can screen for PGC 1alpha stimulation. We chose PGC 1alpha since this is one of the major target whose stimulation has been shown to directly affect antiaging and metabolic outcomes. In a routine screen of botanical extracts we found that Lippia Sidoides can stimulate PGC 1alpha expression. Lippia (Verbenaceae) comprises a number of medicinal and economically important species, including several that are used in traditional Brazilian medicine. We also found that it activates cAMP response element (CRE) in a dose responsive manner. This discovery led us to conclude that Lippia is activating PGC 1alpha via cyclic AMP/PKA signaling. Next we asked the question, is Lippia stimulating cAMP? To answer this question, we used IBMX which is a non-specific competitive cAMP- and cGMP-dependent phosphodiesterase inhibitor known to raise intracellular cAMP levels by blocking the breakdown of cAMP. We also used Forskolin, a botanical diterpene from Coleus forskohlii, which directly stimulates adenylate cyclase to increase cAMP levels. When combined, forskolin and IBMX synergize to increase intracellular cAMP levels via two different mechanisms. We used IBMX to evaluate Lippia's effects on CRE activation and show that IBMX and Lippia activate CRE in an additive manner. We conclude that Lippia and IBMX appear to be affecting the same mechanism to increase cAMP. Next we tried to determine if Lippia inhibits phosphodiesterases. By a direct enzyme assay we could show that Lippia inhibited cAMP- and cGMP-dependent phosphodiesterases in a dose dependent manner with an IC50 of 3 and 25 ug/ml, respectively. The above data leads to the conclusion that Lippia sidoides stimulates mitochondrial biogenesis due to its Phosphodiestrase activity.