Pretreatment of Exosomes Derived from hUCMSCs with LPS Ameliorates Liver Fibrosis by Inhibiting the Smad3C Through Upregulating MiR-126 in the HSC-T6 Cell Line

IF 0.3 4区医学 Q4 GASTROENTEROLOGY & HEPATOLOGY

Hepatitis Monthly Pub Date : 2023-12-02 DOI:10.5812/hepatmon-137083

Maryam Cheraghzadeh, Emad Matour, Benyamin Kabizadeh, Elham Shakerian, Mojtaba Rashidi

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引用次数: 0

Abstract

Background: Activated hepatic stellate cells (HSC) through the TGF-β signaling pathway are likely to exacerbate liver fibrosis, which is a later stage of NASH, a condition in which, in addition to HSC activation, the overexpression of extracellular matrix (ECM) proteins, specifically collagen-I α and α-SMA, is expected. MicroRNA-126a, a modulator of HSC activation, influences the phosphorylation of Smad2/3. Objectives: This study aimed to investigate the impact of exosomes on miRNAs implicated in the progression of liver fibrosis by focusing on the role of miR-126a in the HSC-T6 cell line. Methods: In the present study, we investigated the effects of exosomes derived from LPS-stimulated mesenchymal stem cells (MSCs) on the expression of miR-126a and the phosphorylation of Smad3c in the HSC-T6 cell line. First, HSC-T6 cells were cultured in DMEM supplemented with 10% FBS. Next, we isolated exosomes derived from MSCs using the ExoCib kit. Finally, we examined the gene expression levels of collagen-Iα, α-SMA, and miR-126a using real-time PCR. Results: The results showed that treatment with TGFβ1 increased the phosphorylation of p-Smad3c (P < 0.0001), as well as the expression of α-SMA and Collagen-Iα. Conversely, miR-126a expression was down-regulated after treatment with TGFβ1 (P < 0.01). However, exposure to LPS-treated MSC-derived exosomes resulted in a significant decrease in the levels of p-Smad3c phosphorylation (P < 0.001) and the gene expression of α-SMA and Collagen-Iα, accompanied by the up-regulation of miR-126a (P < 0.05). Conclusions: Based on our observations, MSCs-derived exosomes were able to reduce the expression of the genes associated with hepatic fibrosis, such as collagen-Iα and α-SMA. Furthermore, exosomes inhibited Smad3c phosphorylation by increasing miR-126a expression, ultimately hindering the progression of liver fibrosis. Therefore, exosomes should be recognized as a valuable and beneficial therapeutic tool for liver fibrosis.

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用 LPS 预处理 hUCMSCs 外泌体可通过上调 MiR-126 抑制 HSC-T6 细胞系中的 Smad3C 来改善肝纤维化

背景:通过TGF-β信号通路激活的肝星状细胞(HSC)可能加剧肝纤维化，这是NASH的后期阶段，在这种情况下，除了HSC激活外，细胞外基质(ECM)蛋白，特别是胶原- i α和α- sma的过度表达是预期的。MicroRNA-126a是HSC活化的调节因子，影响Smad2/3的磷酸化。目的:本研究旨在通过关注miR-126a在HSC-T6细胞系中的作用来研究外泌体对涉及肝纤维化进展的mirna的影响。方法:在本研究中，我们研究了来自lps刺激的间充质干细胞(MSCs)的外泌体对HSC-T6细胞系中miR-126a表达和Smad3c磷酸化的影响。首先，将HSC-T6细胞培养在添加10%胎牛血清的DMEM中。接下来，我们使用ExoCib试剂盒分离来自MSCs的外泌体。最后，我们使用实时PCR检测了胶原- i α、α-SMA和miR-126a的基因表达水平。结果:tgf - β1处理后，P - smad3c磷酸化水平升高(P < 0.0001)， α-SMA和胶原- i α表达升高。相反，tgf - β1处理后miR-126a表达下调(P < 0.01)。然而，暴露于lps处理的msc来源的外泌体导致P - smad3c磷酸化水平显著降低(P < 0.001)， α-SMA和胶原- i - α基因表达显著降低，同时miR-126a上调(P < 0.05)。结论:根据我们的观察，间质干细胞衍生的外泌体能够降低与肝纤维化相关的基因的表达，如胶原- i α和α-SMA。此外，外泌体通过增加miR-126a的表达来抑制Smad3c磷酸化，最终阻碍肝纤维化的进展。因此，外泌体应该被认为是一种有价值和有益的肝纤维化治疗工具。

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

Hepatitis Monthly 医学-胃肠肝病学

CiteScore

1.50

自引率

0.00%

发文量

审稿时长

3 months

期刊介绍： Hepatitis Monthly is a clinical journal which is informative to all practitioners like gastroenterologists, hepatologists and infectious disease specialists and internists. This authoritative clinical journal was founded by Professor Seyed-Moayed Alavian in 2002. The Journal context is devoted to the particular compilation of the latest worldwide and interdisciplinary approach and findings including original manuscripts, meta-analyses and reviews, health economic papers, debates and consensus statements of the clinical relevance of hepatological field especially liver diseases. In addition, consensus evidential reports not only highlight the new observations, original research, and results accompanied by innovative treatments and all the other relevant topics but also include highlighting disease mechanisms or important clinical observations and letters on articles published in the journal.