Novel Fluorescence-Based Method To Characterize the Antioxidative Effects of Food Metabolites on Lipid Droplets in Cultured HepatocytesClick to copy article linkArticle link copied!
- Takayuki TsukuiTakayuki TsukuiDepartment of Nutrition, Sapporo University of Health Sciences, Nakanuma Nishi-4-3-1-15, Higashi-ku, Sapporo 007-0894, JapanMore by Takayuki Tsukui
- Zhen ChenZhen ChenFaculty of Health Sciences, Hokkaido University, Kita-12, Nishi-5, Kita-ku, Sapporo 060-0812, JapanMore by Zhen Chen
- Hirotoshi FudaHirotoshi FudaFaculty of Health Sciences, Hokkaido University, Kita-12, Nishi-5, Kita-ku, Sapporo 060-0812, JapanMore by Hirotoshi Fuda
- Takayuki FurukawaTakayuki FurukawaFaculty of Health Sciences, Hokkaido University, Kita-12, Nishi-5, Kita-ku, Sapporo 060-0812, JapanMore by Takayuki Furukawa
- Kotaro OuraKotaro OuraFaculty of Health Sciences, Hokkaido University, Kita-12, Nishi-5, Kita-ku, Sapporo 060-0812, JapanMore by Kotaro Oura
- Toshihiro SakuraiToshihiro SakuraiFaculty of Health Sciences, Hokkaido University, Kita-12, Nishi-5, Kita-ku, Sapporo 060-0812, JapanMore by Toshihiro Sakurai
- Shu-Ping Hui*Shu-Ping Hui*E-mail: [email protected]. Phone & Fax: +81-11-706-3693.Faculty of Health Sciences, Hokkaido University, Kita-12, Nishi-5, Kita-ku, Sapporo 060-0812, JapanMore by Shu-Ping Hui
- Hitoshi ChibaHitoshi ChibaDepartment of Nutrition, Sapporo University of Health Sciences, Nakanuma Nishi-4-3-1-15, Higashi-ku, Sapporo 007-0894, JapanMore by Hitoshi Chiba
Abstract
A fluorescence microscopic method for characterizing size, quantity, and oxidation of lipid droplets (LDs) in HepG2 cells was developed. LDs were induced by palmitic (PA), oleic (OA), or linoleic acids (LA) and stained with two fluorescent probes for neutral lipids and lipid peroxides. Each fatty acid increased the number of LDs and oxidized LDs (oxLDs) and the degree of LD oxidation time dependently, as well as increased intracellular triglyceride hydroperoxides. LDs induced by LA without 2,2′-azobis(2-amidinopropane)dihydrochloride (AAPH) showed the most significant oxidation degree over PA and OA, especially in large LDs (area ≥ 3 μm2, oxLD/LD = 52.3 ± 21.7%). Under this condition, two food-derived antioxidants were evaluated, and both of them significantly improved the LD characteristics. Moreover, chlorogenic acid reduced the quantity of large LDs by 74.0–87.6% in a dose-dependent manner. The proposed method provides a new approach to evaluate the effect of dietary antioxidants on LD characteristics.
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