Bioactive compounds reported to stimulate mitochondrial biogenesis are linked to many health benefits such increased longevity, improved energy utilization, and protection from reactive oxygen species. Previously studies have shown that mice and rats fed diets lacking in pyrroloquinoline quinone (PQQ) have reduced mitochondrial content. Therefore, we hypothesized that PQQ can induce mitochondrial biogenesis in mouse hepatocytes. Exposure of mouse Hepa1-6 cells to 10 -30 M PQQ for 24 -48 h resulted in increased citrate synthase and cytochrome c oxidase activity, Mitotracker staining, mitochondrial DNA content, and cellular oxygen respiration. The induction of this process occurred through the activation of cAMP response element-binding protein (CREB) and peroxisome proliferator-activated receptor-␥ coactivator-1␣ (PGC-1␣), a pathway known to regulate mitochondrial biogenesis. PQQ exposure stimulated phosphorylation of CREB at serine 133, activated the promoter of PGC-1␣, and increased PGC-1␣ mRNA and protein expression. PQQ did not stimulate mitochondrial biogenesis after small interfering RNA-mediated reduction in either PGC-1␣ or CREB expression. Consistent with activation of the PGC-1␣ pathway, PQQ increased nuclear respiratory factor activation (NRF-1 and NRF-2) and Tfam, TFB1M, and TFB2M mRNA expression. Moreover, PQQ protected cells from mitochondrial inhibition by rotenone, 3-nitropropionic acid, antimycin A, and sodium azide. The ability of PQQ to stimulate mitochondrial biogenesis accounts in part for action of this compound and suggests that PQQ may be beneficial in diseases associated with mitochondrial dysfunction.Bioactive compounds, such as pyrroloquinoline quinone (PQQ), 2 resveratrol, genistein, hydroxy-tyrosol, and quercetin have been reported to improve mitochondrial respiratory control or stimulate mitochondrial biogenesis (1-5), which is potentially important to a number of health-related issues ranging from increased longevity, improved energy utilization, and protection from reactive oxygen species (6 -8). Furthermore, mitochondrial DNA depletion and mutations are associated with cardiomyopathy, lactic acidosis, developmental delay, failure to thrive, and impaired neurological function (9). The response to most biofactors is observed after pharmacological intervention or dietary supplementation, although often neargram amounts per kg of diet, or millimolar quantities, are needed for such responses in vivo. PQQ stimulates mitochondriogenesis with the addition of only milligram quantities of PQQ per kg of diet, or micromolar concentrations, in vivo. For example, PQQ deprivation depresses mitochondrial function, which is reversed when as little as 200 -300 g of PQQ/kg of diet are added (1, 10). PQQ also remains detectable in tissues when there is no or little dietary exposure (11), which has not been observed for other dietary polyphenolic compounds known to promote mitochondriogenesis.Recently, PQQ produced by rhizobacterium has been identified as an important plant growth factor (12) and is a...