KDM4C represses liver fibrosis by regulating H3K9me3 methylation of ALKBH5 and m6A methylation of snail1 mRNA

IF 2.3 3区医学 Q3 GASTROENTEROLOGY & HEPATOLOGY

Journal of Digestive Diseases Pub Date : 2024-06-27 DOI:10.1111/1751-2980.13291

Hua Ying Zhou, Bing Qing Wang, Meng Xuan Chen, Yi Fan Wang, Yong Fang Jiang, Jing Ma

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引用次数: 0

Abstract

Objective

We aimed to disclose the molecular mechanism of snail1 in liver fibrosis.

Methods

Carbon tetrachloride (CCl₄) was used to induce a liver fibrosis model in mice whereby serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were evaluated, and liver pathological alternations were assessed. Rat hepatic stellate cells (HSC-T6) were irritated with transforming growth factor (TGF)-β1, followed by assessment of cell viability and migration. The levels of snail1, ALKBH5, and lysine specific demethylase 4C (KDM4C) were quantified by immunohistochemistry, western blot, or reverse transcription-quantitative polymerase chain reaction, in addition to α-smooth muscle actin (SMA), anti-collagen type I α1 (COL1A1), vimentin, and E-cadherin. Photoactivatable ribonucleoside-enhanced crosslinking and immunoprecipitation and RNA stability were evaluated to determine the relationship between ALKBH5 and snail1. Changes in KDM4C-bound ALKBH5 promoter and enrichment of histone H3 lysine 9 trimethylation (H3K9me3) at the ALKBH5 promoter were determined using chromatin immunoprecipitation.

Results

In fibrosis mice, snail1 was upregulated while ALKBH5 and KDM4C were downregulated. KDM4C overexpression reduced serum ALT and AST levels, liver injury, and α-SMA, COL1A1 and VIMENTIN expressions but increased E-cadherin expression. However, the aforementioned trends were reversed by concurrent overexpression of snail1. In HSC-T6 cells exposed to TGF-β1, ALKBH5 overexpression weakened cell viability and migration, downregulated α-SMA, COL1A1 and VIMENTIN, upregulated E-CADHERIN, and decreased m6A modification of snail1 and its mRNA stability. KDM4C increased ALKBH5 expression by lowering H3K9me3 level, but inhibited HSC-T6 cell activation by regulating the ALKBH5/snail1 axis.

Conclusion

KDM4C decreases H3K9me3 methylation to upregulate ALKBH5 and subsequently inhibits snail1, ultimately impeding liver fibrosis.

查看原文本刊更多论文

KDM4C通过调节ALKBH5的H3K9me3甲基化和蜗牛1 mRNA的m6A甲基化抑制肝纤维化。

目的：揭示蜗牛1在肝纤维化中的分子机制：旨在揭示蜗牛1在肝纤维化中的分子机制：用四氯化碳（CCl4）诱导小鼠肝纤维化模型，评估血清丙氨酸氨基转移酶（ALT）和天门冬氨酸氨基转移酶（AST）水平，并评估肝脏病理变化。用转化生长因子（TGF）-β1刺激大鼠肝星状细胞（HSC-T6），然后评估细胞活力和迁移。除了α-平滑肌肌动蛋白（SMA）、抗Ⅰ型胶原α1（COL1A1）、波形蛋白和E-粘连蛋白外，还通过免疫组化、Western印迹或逆转录定量聚合酶链反应对蜗牛1、ALKBH5和赖氨酸特异性去甲基化酶4C（KDM4C）的水平进行了量化。对光活化核糖核苷增强交联和免疫沉淀以及 RNA 稳定性进行了评估，以确定 ALKBH5 与蜗牛 1 之间的关系。使用染色质免疫沉淀法测定了KDM4C结合的ALKBH5启动子的变化以及组蛋白H3赖氨酸9三甲基化（H3K9me3）在ALKBH5启动子处的富集：结果：在纤维化小鼠中，蜗牛1上调，而ALKBH5和KDM4C下调。KDM4C的过表达降低了血清ALT和AST水平、肝损伤以及α-SMA、COL1A1和VIMENTIN的表达，但增加了E-cadherin的表达。然而，同时过表达蜗牛1会逆转上述趋势。在暴露于 TGF-β1 的 HSC-T6 细胞中，ALKBH5 的过表达削弱了细胞的活力和迁移，下调了 α-SMA、COL1A1 和 VIMENTIN，上调了 E-CADHERIN，并降低了蜗牛 1 的 m6A 修饰及其 mRNA 的稳定性。KDM4C通过降低H3K9me3水平增加了ALKBH5的表达，但通过调节ALKBH5/蜗牛1轴抑制了HSC-T6细胞的活化：结论：KDM4C通过降低H3K9me3甲基化来上调ALKBH5，进而抑制蜗牛1，最终阻碍肝纤维化。

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

Journal of Digestive Diseases 医学-胃肠肝病学

CiteScore

5.40

自引率

2.90%

发文量

审稿时长

6-12 weeks

期刊介绍： The Journal of Digestive Diseases is the official English-language journal of the Chinese Society of Gastroenterology. The journal is published twelve times per year and includes peer-reviewed original papers, review articles and commentaries concerned with research relating to the esophagus, stomach, small intestine, colon, liver, biliary tract and pancreas.