Inhibition of Differentiation of 3T3-L1 Cells by Increasing Glioma-Associated Oncogene Expression in Chrysanthemum indicum L. Using Lactococcus lactis KCTC 3115.

The inhibitory effect of Chrysanthemum indicum L. on adipocyte differentiation can be enhanced by lactic acid bacteria (LAB) fermentation. In this study, we assessed the cellulose resolution, C. indicum L. quantity, and fermentation time and process to verify the LAB selection and fermentation efficiency. In addition, the antioxidant activity, adipocyte signaling and differentiation, and hedgehog (Hh) signaling were investigated, and the changes in compounds before and after fermentation were determined by ultra-high performance liquid chromatography (UHPLC). All strains exhibited satisfactory cellulose resolution. With 20% C. indicum L., fermentation was only effective up to 24 h. The results of the antioxidant assays showed that the 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical scavenging capacities were higher in all fermentations than in unfermented C. indicum L. extract (CI). 3T3-L1 cell differentiation signaling evaluation revealed that CI inhibited adipocyte differentiation by reducing peroxisome proliferator-activated receptor-γ, CCAAT/enhancer binding protein-α, and phosphorylated AMP-activated protein kinase activity in all fermentations. In the Hh signaling analysis, CI fermented with Lactococcus lactis KCTC 3115 significantly increased glioma-associated oncogene 1 (GLI1) activity by inhibiting patched 1 activity and activating smoothened (P<0.001). UHPLC quantitative analysis revealed elevated levels of luteolin and quercetin. Fermentation with C. indicum L. and L. lactis KCTC 3115 activated GLI1, a transcription factor in the Hh signaling pathway, which enhanced the inhibition of adipocyte differentiation, indicating its potential in anti-obesity treatment. However, the exact compounds affecting GLI1 activity require further elucidation in future studies.