杨梅素对长链脂肪酸刺激的原代犊牛肝细胞脂质代谢的影响。

Effect of Myricetin on Lipid Metabolism in Primary Calf Hepatocytes Challenged with Long-Chain Fatty Acids.

作者信息

Yang Wei, Yang Mingmao, Tian Yan, Jiang Qianming, Loor Juan J, Cao Jie, Wang Shuang, Gao Changhong, Fan Wenwen, Zhang Bingbing, Xu Chuang

机构信息

College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China.

Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, College of Veterinary Medicine, Northwest A & F University, Xianyang 712100, China.

出版信息

Metabolites. 2022 Nov 5;12(11):1071. doi: 10.3390/metabo12111071.

Triacylglycerol (TAG) accumulation and oxidative damage in hepatocytes induced by high circulating concentrations of fatty acids (FA) are common after calving. In order to clarify the role of myricetin on lipid metabolism in hepatocytes when FA metabolism increases markedly, we performed in vitro analyses using isolated primary calf hepatocytes from three healthy female calves (1 d old, 42 to 48 kg). Two hours prior to an FA challenge (1.2 mM mix), the hepatocytes were treated with 100 μM (M1), 50 μM (M2), or 25 μM (M3) of myricetin. Subsequently, hepatocytes from each donor were challenged with or without FA for 12 h in an attempt to induce metabolic stress. Data from calf hepatocyte treatment comparisons were assessed using two-way repeated-measures (RM) ANOVA with subsequent Bonferroni correction. The data revealed that hepatocytes challenged with FA had greater concentrations of TAG and nonesterified fatty acids (NEFA), oxidative stress-related MDA and H2O2, and mRNA and protein abundance of lipid synthesis-related SREBF1 and inflammatory-related NF-κB. In addition, the mRNA abundance of the lipid synthesis-related genes FASN, DGAT1, DGAT2, and ACC1; endoplasmic reticulum stress-related GRP79 and PERK; and inflammatory-related TNF-α also were upregulated. In contrast, the activity of antioxidant SOD (p < 0.01) and concentrations of GSH (p < 0.05), and the protein abundance of mitochondrial FA oxidation-related CPT1A, were markedly lower. Compared with FA challenge, 50 and 100 μM myricetin led to lower concentrations of TAG, NEFA, MDA, and H2O2, as well as mRNA and protein abundance of SREBF1, DGAT1, GRP78, and NF-κB. In contrast, the activity of SOD (p < 0.01) and mRNA and protein abundance of CPT1A were markedly greater. Overall, the results suggest that myricetin could enhance the antioxidant capacity and reduce lipotoxicity, endoplasmic reticulum stress, and inflammation. All of these effects can help reduce TAG accumulation in hepatocytes.

产犊后，循环中高浓度脂肪酸（FA）诱导的肝细胞中三酰甘油（TAG）积累和氧化损伤很常见。为了阐明杨梅素在FA代谢显著增加时对肝细胞脂质代谢的作用，我们使用从三头健康雌性犊牛（1日龄，42至48千克）分离的原代犊牛肝细胞进行了体外分析。在FA刺激（1.2 mM混合物）前两小时，肝细胞用100 μM（M1）、50 μM（M2）或25 μM（M3）的杨梅素处理。随后，来自每个供体的肝细胞在有或没有FA的情况下接受12小时刺激，以诱导代谢应激。使用双向重复测量（RM）方差分析并随后进行Bonferroni校正来评估犊牛肝细胞处理比较的数据。数据显示，用FA刺激的肝细胞具有更高浓度的TAG和非酯化脂肪酸（NEFA）、氧化应激相关的丙二醛（MDA）和过氧化氢（H2O2），以及脂质合成相关的固醇调节元件结合蛋白1（SREBF1）和炎症相关的核因子κB（NF-κB）的mRNA和蛋白质丰度。此外，脂质合成相关基因脂肪酸合酶（FASN）、二酰甘油酰基转移酶1（DGAT1）、二酰甘油酰基转移酶2（DGAT2）和乙酰辅酶A羧化酶1（ACC1）；内质网应激相关的葡萄糖调节蛋白79（GRP79）和蛋白激酶R样内质网激酶（PERK）；以及炎症相关的肿瘤坏死因子-α（TNF-α）的mRNA丰度也上调。相比之下，抗氧化超氧化物歧化酶（SOD）的活性（p < 0.01）和谷胱甘肽（GSH）的浓度（p < 0.05），以及线粒体FA氧化相关的肉碱棕榈酰转移酶1A（CPT1A）的蛋白质丰度明显较低。与FA刺激相比，50和100 μM的杨梅素导致TAG、NEFA、MDA和H2O2的浓度降低，以及SREBF1、DGAT1、葡萄糖调节蛋白78（GRP78）和NF-κB的mRNA和蛋白质丰度降低。相比之下，SOD的活性（p < 0.01）以及CPT1A的mRNA和蛋白质丰度明显更高。总体而言，结果表明杨梅素可以增强抗氧化能力并降低脂毒性、内质网应激和炎症。所有这些作用都有助于减少肝细胞中TAG的积累。