Shirouchi Bungo, Yanagi Shuhei, Okawa Chinami, Koga Maiko, Sato Masao
Laboratory of Nutrition Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395, Japan.
Cytotechnology. 2020 Feb;72(1):175-187. doi: 10.1007/s10616-019-00368-5. Epub 2020 Jan 13.
Nuclear receptors, such as liver X receptors (LXRs) and sterol regulatory element-binding proteins (SREBPs), are key regulators of lipogenic genes, including fatty acid synthase (FASN). It has been reported that several oxycholesterols (OCs) act as LXR ligands; however, it is unclear whether all OC molecular species act as ligands. We previously demonstrated that the absorption rate of dietary 6-ketocholestanol (6-keto), an oxycholesterol, is the highest of all the OCs using thoracic lymph duct-cannulated rats. However, limited information is available about the physiological significance of 6-keto. In this study, we investigated whether treatment with 6-keto increases intracellular triacylglycerol (TAG) levels through up-regulation of lipogenic gene expression in HepG2 cells. 6-Keto treatment significantly reduced intracellular TAG levels through down-regulation of lipogenic genes including FASN. Although 6-keto significantly suppressed FASN gene promoter activities, the action was completely diminished when mutations were present in the SREBP promoter site. TO901317 (TO) significantly increased FASN gene promoter activities, whereas simultaneous treatment with TO and 6-keto significantly reduced this activity. We also compared the effects of several OCs that are oxidized at the carbon-6 and -7 in the B-ring of cholesterol on FASN gene promoter activities. Similar to 6-keto, 6α-OH and 6β-OH significantly reduced the activity of the FASN gene promoter, which suggests that oxidation of carbon-6 in the B-ring may play an important role in the reduction of FASN expression. Our results indicate that 6-keto suppresses lipid accumulation by decreasing FASN gene expression through SREBP-dependent regulation in HepG2 cells.
核受体,如肝脏X受体(LXRs)和固醇调节元件结合蛋白(SREBPs),是包括脂肪酸合酶(FASN)在内的生脂基因的关键调节因子。据报道,几种氧化胆固醇(OCs)可作为LXR配体;然而,尚不清楚是否所有的OC分子种类都作为配体。我们之前使用胸导管插管大鼠证明,饮食中的氧化胆固醇6-酮胆甾烷醇(6-酮)的吸收率在所有OCs中是最高的。然而,关于6-酮的生理意义的信息有限。在本研究中,我们研究了用6-酮处理是否通过上调HepG2细胞中生脂基因的表达来增加细胞内三酰甘油(TAG)水平。6-酮处理通过下调包括FASN在内的生脂基因显著降低了细胞内TAG水平。尽管6-酮显著抑制FASN基因启动子活性,但当SREBP启动子位点存在突变时,该作用完全消失。TO901317(TO)显著增加FASN基因启动子活性,而TO和6-酮同时处理则显著降低该活性。我们还比较了几种在胆固醇B环的碳6和碳7处被氧化的OCs对FASN基因启动子活性的影响。与6-酮类似,6α-OH和6β-OH显著降低了FASN基因启动子的活性,这表明B环中碳6的氧化可能在降低FASN表达中起重要作用。我们的结果表明,6-酮通过SREBP依赖性调节降低HepG2细胞中FASN基因表达来抑制脂质积累。