Curcumin Modulates NOX Gene Expression and ROS Production via P-Smad3C in TGF-β-Activated Hepatic Stellate Cells.

Q2 Biochemistry, Genetics and Molecular Biology
Iranian Biomedical Journal Pub Date : 2024-01-01 Epub Date: 2023-10-28 DOI:10.61186/ibj.4005
Shahla Asadizade, Mahdi Hatami, Samaneh Salehipour Bavarsad, Benyamin Kabizade, Elham Shakerian, Mojtaba Rashidi
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引用次数: 0

Abstract

Background: Liver fibrosis, associated with hepatic stellate cells (HSCs), occurs when a healthy liver sustains damage, thereby impairing its function. NADPH oxidases (NOXs), specifically isoforms 1, 2, and 4, play a role in reactive oxygen species (ROS) production during hepatic injuries, resulting in fibrosis. Curcumin has shown strong potential in mitigating liver fibrosis. Our research aimed to investigate the effects of curcumin on lowering NOX and ROS levels. This compound was also studied for its effects on NOXs, ROS concentrations through the inhibition of Smad3 phosphorylation in transforming growth factor beta (TGF-β)-activated human HSCs.

Methods: MTT assay investigated the cytotoxic effects of curcumin on HSCs. The cells were activated by exposure to TGF-β (2 ng/mL) for 24 hours. After activating, the cells were treated with curcumin at 25-150 μM concentrations. After administering curcumin to the cells, we employed RT-PCR and Western blot techniques to evaluate the related gene and protein expression levels. This evaluation was primarily focused on the mRNA expression levels of NOX1, NOX2, NOX4 and phosphorylated Smad3C.

Results: The mRNA expression level of aforesaid NOXs as well as α-smooth muscle actin (α-SMA), collagen1-α, and ROS levels were significantly reduced following 100 μM curcumin treatment. Furthermore, curcumin significantly decreased the p-Smad3C protein level in TGF-β-activated cells, with fold changes of 3 and 2 observed at 75 and 100 μM, respectively.

Conclusion: Curcumin decreased the levels of ROS and NOX, as well as the expression of α-SMA and collagen1-α. The primary mechanism for this reduction could be linked to the level of p-Smad3C. Hence, curcumin could serve as an effective therapeutic agent for liver fibrosis.

姜黄素通过 P-Smad3C 调节 TGF-β 激活的肝星状细胞中 NOX 基因的表达和 ROS 的产生
背景:肝纤维化与造血干细胞有关:肝纤维化与造血干细胞有关,当健康的肝脏受到损伤,其功能受损时就会发生肝纤维化。NOX(1、2 和 4)在损伤过程中产生 ROS,导致肝纤维化。本研究旨在探讨姜黄素在降低 NOX 和 ROS 水平方面的作用。本研究还研究了姜黄素对 TGF-β 激活的人造血干细胞中 NOXs、ROS 生成和 Smad3 磷酸化的影响:方法:采用 MTT 法研究姜黄素对造血干细胞的细胞毒性作用。将细胞暴露于 TGF-β(2 ng/mL)24 小时后激活。激活后,用不同浓度(25-150 μM)的姜黄素处理细胞。给细胞注射姜黄素后,使用 RT-PCR 和 Western 印迹技术对相关基因和蛋白质的表达水平进行了评估。评估主要集中在 NOX1、NOX2、NOX4 和磷酸化 Smad3C 的 mRNA 表达水平上:结果表明,姜黄素浓度为 100 μM 时,NOX1、NOX2 和 NOX4 的 mRNA 表达水平以及 α-SMA、胶原蛋白 1-α 和 ROS 水平均明显降低。此外,姜黄素还能明显降低 TGF-β 激活细胞中 p-Smad3C 蛋白水平,在 75 μM 和 100 μM 浓度下分别观察到 3 倍和 2 倍的变化:姜黄素能降低 ROS 和 NOX 水平,以及 α-SMA 和胶原蛋白 1-α 的表达。这种降低的主要机制可能与 p-Smad3C 的水平有关。因此,姜黄素有可能被视为治疗肝纤维化的有效药物。
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
Iranian Biomedical Journal
Iranian Biomedical Journal Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
3.20
自引率
0.00%
发文量
42
审稿时长
8 weeks
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