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绿茶多酚(-)-表儿茶素预处理可减轻体外MAFLD模型中的肝脂肪变性。

Green Tea Polyphenol (-)-Epicatechin Pretreatment Mitigates Hepatic Steatosis in an In Vitro MASLD Model.

作者信息

Hefer Marija, Petrovic Ana, Roguljic Lucija Kuna, Kolaric Tea Omanovic, Kizivat Tomislav, Wu Catherine H, Tabll Ashraf A, Smolic Robert, Vcev Aleksandar, Smolic Martina

机构信息

Department of Translational Medicine, Faculty of Dental Medicine and Health Osijek, J. J. Strossmayer University of Osijek, 31000 Osijek, Croatia.

Department of Pharmacology and Biochemistry, Faculty of Dental Medicine and Health Osijek, J. J. Strossmayer University of Osijek, 31000 Osijek, Croatia.

出版信息

Curr Issues Mol Biol. 2024 Aug 16;46(8):8981-8994. doi: 10.3390/cimb46080531.

Abstract

Metabolic dysfunction-associated steatotic liver disease (MASLD), previously known as non-alcoholic fatty liver disease (NAFLD), is becoming more prominent globally due to an increase in the prevalence of obesity, dyslipidemia, and type 2 diabetes. A great deal of studies have proposed potential treatments for MASLD, with few of them demonstrating promising results. The aim of this study was to investigate the potential effects of (-)-epicatechin (EPI) on the development of MASLD in an in vitro model using the HepG2 cell line by determining the metabolic viability of the cells and the levels of PPARα, PPARγ, and GSH. HepG2 cells were pretreated with 10, 30, 50, and 100 μM EPI for 4 h to assess the potential effects of EPI on lipid metabolism. A MASLD cell culture model was established using HepG2 hepatocytes which were exposed to 1.5 mM oleic acid (OA) for 24 h. Moreover, colorimetric MTS assay was used in order to determine the metabolic viability of the cells, PPARα and PPARγ protein levels were determined using enzyme-linked immunosorbent assay (ELISA), and lipid accumulation was visualized using the Oil Red O Staining method. Also, the levels of intracellular glutathione (GSH) were measured to determine the level of oxidative stress. EPI was shown to increase the metabolic viability of the cells treated with OA. The metabolic viability of HepG2 cells, after 24 h incubation with OA, was significantly decreased, with a metabolic viability of 71%, compared to the cells pretreated with EPI, where the metabolic viability was 74-86% with respect to the concentration of EPI used in the experiment. Furthermore, the levels of PPARα, PPARγ, and GSH exhibited a decrease in response to increasing EPI concentrations. Pretreatment with EPI has demonstrated a great effect on the levels of PPARα, PPARγ, and GSH in vitro. Therefore, considering that EPI mediates lipid metabolism in MASLD, it should be considered a promising hepatoprotective agent in future research.

摘要

代谢功能障碍相关脂肪性肝病(MASLD),以前称为非酒精性脂肪性肝病(NAFLD),由于肥胖、血脂异常和2型糖尿病患病率的增加,在全球范围内正变得越来越突出。大量研究提出了针对MASLD的潜在治疗方法,但其中很少有显示出有前景的结果。本研究的目的是通过测定细胞的代谢活力以及PPARα、PPARγ和谷胱甘肽(GSH)的水平,在使用HepG2细胞系的体外模型中研究(-)-表儿茶素(EPI)对MASLD发展的潜在影响。用10、30、50和100μM EPI预处理HepG2细胞4小时,以评估EPI对脂质代谢的潜在影响。使用暴露于1.5 mM油酸(OA)24小时的HepG2肝细胞建立MASLD细胞培养模型。此外,采用比色MTS法测定细胞的代谢活力,使用酶联免疫吸附测定(ELISA)法测定PPARα和PPARγ蛋白水平,并使用油红O染色法观察脂质蓄积情况。同时,测量细胞内谷胱甘肽(GSH)的水平以确定氧化应激水平。结果显示,EPI可提高用OA处理的细胞代谢活力。与用EPI预处理的细胞相比,OA孵育24小时后HepG2细胞代谢活力显著降低,代谢活力为71%,而根据实验中所用EPI的浓度,用EPI预处理的细胞代谢活力为74%-86%。此外,PPARα、PPARγ和GSH水平随着EPI浓度的增加而降低。EPI预处理已在体外对PPARα、PPARγ和GSH水平显示出显著影响。因此,考虑到EPI在MASLD中介导脂质代谢,在未来研究中应将其视为一种有前景的肝保护剂。

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