Gardenin A alleviates alcohol-induced oxidative stress and inflammation in HepG2 and Caco2 cells via AMPK/Nrf2 pathway

IF 4.5 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Bioorganic Chemistry Pub Date : 2025-04-28 DOI:10.1016/j.bioorg.2025.108543

Prashsti Chadha , Hiral Aghara , Delna Johnson , Dhrubjyoti Sharma , Mitalben Odedara , Manali Patel , Hemant Kumar , Vijay Thiruvenkatam , Palash Mandal

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Abstract

Chronic alcohol consumption triggers immune responses that lead to cell damage, contributing to alcohol-associated liver disease (ALD). Despite its prevalence, no FDA-approved treatment for ALD currently exists. This study explores the cytoprotective effects of Gardenin A (GarA), a polymethoxylated flavone, for protection against alcohol-induced oxidative stress and inflammation in HepG2 and Caco2 cell lines. GarA was isolated, characterized and, tested in-vitro, showing maximum cell viability at 10 μg/ml using MTT assays. Further, lipid accumulation assay, reactive oxygen species (ROS) estimation and nuclear morphology visualization was carried out using different staining techniques. RT-qPCR was employed to examine the expression of various pro- and anti-inflammatory cytokines, along with Cytochrome P4502E1 (CYP2E1) and Sterol regulatory element binding protein-2 (SREBP2) and tight junction genes crucial for gut barrier integrity. Moreover, ELISA was carried out for key protein targets such as AMPK (phosphorylated and total), TNFα, C5aR1, HO-1 and Nrf2. GarA caused a marked decrease in lipid droplets, ROS levels, and expression of pro-inflammatory cytokines. It showed anti-inflammatory and anti-oxidant activity and helped maintain the gut barrier and nuclear integrity. In-silico studies showed the conserved amino acid interaction and affinity of GarA with C5aR1, and TNFα, compared to the interactions with known inhibitors/activators, further corroborating the results. This study is the first to explore the effects of GarA on ALD, underscoring its potential as an anti-inflammatory and anti-oxidant agent targeting the AMPK/Nrf2 signaling pathway, suggesting its future as a promising therapeutic candidate for mitigating alcohol-induced liver and gut damage.

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栀子苷A通过AMPK/Nrf2通路缓解酒精诱导的HepG2和Caco2细胞氧化应激和炎症

长期饮酒会引发免疫反应，导致细胞损伤，导致酒精相关性肝病（ALD）。尽管它很流行，但目前还没有fda批准的治疗ALD的方法。本研究探讨了栀子苷A （GarA）的细胞保护作用，GarA是一种多甲氧基黄酮，对HepG2和Caco2细胞系酒精诱导的氧化应激和炎症具有保护作用。对GarA进行了分离、表征和体外测试，MTT检测显示10 μg/ml时细胞活力最大。此外，脂质积累测定，活性氧（ROS）估计和核形态可视化使用不同的染色技术进行。RT-qPCR检测了各种促炎和抗炎细胞因子、细胞色素P4502E1 （CYP2E1）和甾醇调节元件结合蛋白2 （SREBP2）以及对肠道屏障完整性至关重要的紧密连接基因的表达。此外，对关键蛋白靶点如AMPK（磷酸化和总）、TNFα、C5aR1、HO-1和Nrf2进行ELISA检测。GarA引起脂滴、ROS水平和促炎细胞因子表达的显著降低。它具有抗炎和抗氧化活性，有助于维持肠道屏障和核完整性。与已知的抑制剂/激活剂的相互作用相比，计算机研究表明，GarA与C5aR1和TNFα的氨基酸相互作用和亲和力是保守的，进一步证实了结果。这项研究首次探索了GarA对ALD的影响，强调了其作为靶向AMPK/Nrf2信号通路的抗炎和抗氧化剂的潜力，表明其作为缓解酒精诱导的肝脏和肠道损伤的有前途的治疗候选药物。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bioorganic Chemistry 生物-生化与分子生物学

CiteScore

9.70

自引率

3.90%

发文量

679

审稿时长

31 days

期刊介绍： Bioorganic Chemistry publishes research that addresses biological questions at the molecular level, using organic chemistry and principles of physical organic chemistry. The scope of the journal covers a range of topics at the organic chemistry-biology interface, including: enzyme catalysis, biotransformation and enzyme inhibition; nucleic acids chemistry; medicinal chemistry; natural product chemistry, natural product synthesis and natural product biosynthesis; antimicrobial agents; lipid and peptide chemistry; biophysical chemistry; biological probes; bio-orthogonal chemistry and biomimetic chemistry. For manuscripts dealing with synthetic bioactive compounds, the Journal requires that the molecular target of the compounds described must be known, and must be demonstrated experimentally in the manuscript. For studies involving natural products, if the molecular target is unknown, some data beyond simple cell-based toxicity studies to provide insight into the mechanism of action is required. Studies supported by molecular docking are welcome, but must be supported by experimental data. The Journal does not consider manuscripts that are purely theoretical or computational in nature. The Journal publishes regular articles, short communications and reviews. Reviews are normally invited by Editors or Editorial Board members. Authors of unsolicited reviews should first contact an Editor or Editorial Board member to determine whether the proposed article is within the scope of the Journal.