肝微管不稳定促进肝豆状核变性小鼠模型肝纤维化。

IF 4.8 3区 医学 Q1 GENETICS & HEREDITY
Journal of Molecular Medicine-Jmm Pub Date : 2025-05-01 Epub Date: 2025-03-26 DOI:10.1007/s00109-025-02535-y
Som Dev, Yixuan Dong, James P Hamilton
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引用次数: 0

摘要

威尔逊病(WD)是一种潜在致命的代谢性疾病,由铜(Cu)转运体ATP7B失活引起,导致全身铜超载和纤维性炎症性肝病。高铜对肝纤维化细胞骨架动力学的分子机制和影响尚不清楚。在这里,我们测试了肝细胞骨架和纤维生成的调节与Cu超载在WD中的关系。采用Western blotting、实时定量反转录PCR (qRT-PCR)、免疫组织化学(IHC)染色和转录组学(rna测序)分析,比较已确定肝病的Atp7b-/-(敲除)小鼠、肝细胞特异性Atp7b△Hep敲除小鼠和年龄和性别匹配的对照组。在患有严重肝病的Atp7b-/-小鼠中,细胞骨架蛋白表达显著增加,α-微管蛋白乙酰化降低。在这些小鼠发生肝脏病理之前,没有观察到细胞骨架和肝星状细胞活化的显著变化。随着肝铜水平的升高,细胞骨架蛋白增加,乙酰化-α-微管蛋白/α-微管蛋白比例降低。rna测序、qRT-PCR和免疫染色证实微管蛋白在转录水平上调,肝细胞是纤维化前早期微管蛋白增加的主要来源。通过Hdac6和Sirt2诱导α-微管蛋白乙酰化减少,α-微管蛋白增加,促进纤维化,这反映在20周时Atp7b-/-小鼠的desmin和α-SMA免疫染色同时增加。此外,α-微管蛋白和α-微管蛋白去乙酰化酶与动物和人WD中肝纤维化标志物的表达呈显著正相关。肝细胞特异性Atp7b△Hep小鼠尽管肝脂肪变性,但在微管蛋白和纤维化方面缺乏显著变化。本研究提供了证据,证明微管不稳定导致细胞骨架重排,促进了小鼠WD模型中肝星状细胞(HSC)的激活和纤维化。关键信息:肝豆状核变性诱导肝细胞骨架系统。肝微管乙酰化在小鼠威尔逊病中失调。肝豆状核变性患者的微管不稳定与肝纤维化呈正相关。伴随纤维形成的微管不稳定加剧了WD的进展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Hepatic microtubule destabilization facilitates liver fibrosis in the mouse model of Wilson disease.

Wilson disease (WD) is a potentially fatal metabolic disorder caused by the inactivation of the copper (Cu) transporter ATP7B, resulting in systemic Cu overload and fibroinflammatory liver disease. The molecular mechanism and effects of elevated Cu on cytoskeletal dynamics in liver fibrogenesis are not clear. Here, we tested the regulation of hepatic cytoskeleton and fibrogenesis with respect to Cu overload in WD. Atp7b-/- (knockout) mice with established liver disease, hepatocyte-specific Atp7b△Hep knockout mice without fibroinflammatory disease, and the age-and sex-matched controls were compared using Western blotting, real-time quantitative reverse transcription PCR (qRT-PCR), immunohistochemical (IHC) staining and transcriptomics (RNA-sequencing) analysis. In Atp7b-/- mice with developed liver disease, there is a significant increase in cytoskeletal protein expression with a reduction in α-tubulin acetylation. In these mice before the onset of liver pathology, no significant changes in cytoskeletal nor hepatic stellate cell activation are observed. As hepatic copper levels rise, an increase in cytoskeletal proteins with a decrease in acetylated-α-tubulin/α-tubulin ratio occurs. RNA-sequencing, qRT-PCR, and immunostaining confirm that the tubulin is upregulated at the transcriptional level and hepatocytes are the primary source of early tubulin increases before fibrosis. An increase in α-tubulin with a decrease in α-tubulin acetylation via Hdac6 and Sirt2 induction facilitates fibrosis as reflected by concomitant increases in desmin and α-SMA immunostaining in Atp7b-/- mice at 20 weeks. Moreover, strongly positive correlations between α-tubulin and α-tubulin deacetylase with the expression of liver fibrosis markers are observed in animal and human WD. Hepatocyte-specific Atp7b△Hep mice lack significant changes in tubulin as well as fibrosis despite hepatic steatosis. This study provides evidence that microtubule destabilization causes cytoskeletal rearrangement and facilitates hepatic stellate cell (HSC) activation and fibrosis in the murine model of WD. KEY MESSAGES: Hepatic cytoskeleton system is induced in Wilson disease. Hepatic microtubules acetylation is dysregulated in murine Wilson disease. Microtubules destabilization is positively associated with liver fibrosis in Wilson disease. Microtubules destabilization concomitant with fibrogenesis exacerbates WD progression.

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来源期刊
Journal of Molecular Medicine-Jmm
Journal of Molecular Medicine-Jmm 医学-医学:研究与实验
CiteScore
9.30
自引率
0.00%
发文量
100
审稿时长
1.3 months
期刊介绍: The Journal of Molecular Medicine publishes original research articles and review articles that range from basic findings in mechanisms of disease pathogenesis to therapy. The focus includes all human diseases, including but not limited to: Aging, angiogenesis, autoimmune diseases as well as other inflammatory diseases, cancer, cardiovascular diseases, development and differentiation, endocrinology, gastrointestinal diseases and hepatology, genetics and epigenetics, hematology, hypoxia research, immunology, infectious diseases, metabolic disorders, neuroscience of diseases, -omics based disease research, regenerative medicine, and stem cell research. Studies solely based on cell lines will not be considered. Studies that are based on model organisms will be considered as long as they are directly relevant to human disease.
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