| In Vitro | In vitro activity: Asaraldehyde is found to be the most abundant constituent, but is totally absent in its cultured broth and its natural host, C. kanehirae wood. On feeding with the acid-digested sawdust of C. kanehirae wood or vanillin to the broth for culture, Asaraldehyde is produced in both cultured broths. Asaraldehyde shows 3.32% of prostaglandin H endoperoxide synthase-1 (COX-1) inhibitory activity and 52.69% prostaglandin H endoperoxide synthase-2 (COX-2) inhibitory activity, respectively at 100 mg/mL. Asaraldehyde shows selectivity towards COX-2 enzyme inhibition at 100 μg/mL. The COX-2/COX-1 ratio for Asaraldehyde is 17.68 at 100 μg/mL compared to solvent control. Asaraldehyde down-regulates C/EBPβ, C/EBPδ, and C/EBPα. Asaraldehyde suppresses expression of PPARγ1 and PPARγ2. Asaraldehyde also significantly suppresses the expression of acetyl-CoA carboxylase (ACC).
Cell Assay: 3T3-L1 cells are seeded in 96-well plates at a concentration of 104 /well. Twenty-four hours after seeding, the cells are treated with 100 μg/mL of Asaraldehyde for 24 hours or for the whole 8-day differentiation period. Fully differentiated adipocytes are also treated with 100 μg/mL of Asaraldehyde for 24 hours-72 hours to test the cytotoxicity. At the end of treatment, cells are cultured with MTT at a final concentration of 0.5 mg/mL for another 4 hours. The purple MTT formazan is dissolved by DMSO and the absorbance at 570 nm is taken with a spectrophotometer. The absorbance is proportional to the viability of adipocytes. |
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