Inhibition of GSK3β activity alleviates acute liver failure via suppressing multiple programmed cell death.

IF 4.4 3区医学 Q2 IMMUNOLOGY

Journal of Inflammation-London Pub Date : 2023-07-13 DOI:10.1186/s12950-023-00350-1

Danmei Zhang, Chunxia Shi, Qingqi Zhang, Yukun Wang, Jin Guo, Zuojiong Gong

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引用次数: 0

Abstract

Background: Acute liver failure (ALF) is one of the most common life-threatening diseases in adults without previous liver disease. Glycogen synthase kinase 3β (GSK3β) is a serine/threonine protein kinase that is widely distributed in the cells. Inhibition of its activity can inhibit cell death and promote autophagy through various pathways, thus providing a protective effect. In this study, we aimed to investigate the effect on ALF after inhibition of GSK3β and its potential mechanisms.

Methods: D- galactosamine(D-Gal) in combination with lipopolysaccharide(LPS) was used to induce ALF in vitro and in vivo. And then GSK3β inhibitor TDZD-8 was used to explore the protective effect against ALF. After TDZD-8 treatment TUNEL staining and flow techniques were used to detect the proportion of apoptosis in liver tissues and cells respectively, while western blotting and immunofluorescence assays were performed to detect the expression levels of apoptosis, pyroptosis and necroptosis-related proteins in tissues and cells. In addition, western blotting was performed to explore the specific mechanism of hepatoprotective effect after GSK3β inhibition to detect the expression levels of TAK1, TRAF6 and HDAC3 after TRAF6 and HDAC3 inhibition alone. The co-localization of TRAF6 and HDAC3 in vitro was detected by immunofluorescence, while the interaction between TRAF6 and HDAC3 was detected by immunoprecipitation assay.

Results: Both in vivo and in vitro experiments, GSK3β inhibitor TDZD-8 can significantly alleviate the progression of ALF. Inhibition of GSK3β activity could significantly reduce the level of hepatocyte apoptosis, pyroptosis, necroptosis and improve liver dysfunction and tissue damage. Furthermore, we found that hepatocyte TAK1 and TRAF6 levels decreased and HDAC3 levels increased in ALF, whereas inhibition of GSK3β upregulated TAK1 and TRAF6 levels and decreased HDAC3 expression.

Conclusion: GSK3β inhibitor TDZD-8 can prevent the progression of ALF, and its action may involve the TRAF6/HDAC3/TAK1 pathway.

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抑制 GSK3β 的活性可通过抑制多种程序性细胞死亡缓解急性肝功能衰竭。

背景：急性肝功能衰竭（ALF）是既往无肝病的成年人中最常见的危及生命的疾病之一。糖原合酶激酶 3β（GSK3β）是一种丝氨酸/苏氨酸蛋白激酶，广泛分布于细胞中。抑制其活性可通过多种途径抑制细胞死亡和促进自噬，从而起到保护作用。本研究旨在探讨抑制GSK3β后对ALF的影响及其潜在机制：方法：采用D-半乳糖胺（D-Gal）联合脂多糖（LPS）在体外和体内诱导ALF。然后用 GSK3β 抑制剂 TDZD-8 探讨其对 ALF 的保护作用。TDZD-8治疗后，采用TUNEL染色和流式技术分别检测肝组织和细胞中的凋亡比例，同时采用Western印迹和免疫荧光检测组织和细胞中凋亡、热凋亡和坏死相关蛋白的表达水平。此外，为了探索 GSK3β 抑制后保肝作用的具体机制，还进行了 Western 印迹分析，以检测 TRAF6 和 HDAC3 单独抑制后 TAK1、TRAF6 和 HDAC3 的表达水平。免疫荧光法检测了TRAF6和HDAC3在体外的共定位，免疫沉淀法检测了TRAF6和HDAC3之间的相互作用：结果：在体内和体外实验中，GSK3β抑制剂TDZD-8都能显著缓解ALF的进展。抑制 GSK3β 活性可明显降低肝细胞凋亡、热凋亡和坏死水平，改善肝功能异常和组织损伤。此外，我们还发现，ALF患者肝细胞TAK1和TRAF6水平降低，HDAC3水平升高，而抑制GSK3β可上调TAK1和TRAF6水平，降低HDAC3的表达：结论：GSK3β抑制剂TDZD-8可阻止ALF的进展，其作用可能涉及TRAF6/HDAC3/TAK1通路。

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 Inflammation-London IMMUNOLOGY-

CiteScore

7.90

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Journal of Inflammation welcomes research submissions on all aspects of inflammation. The five classical symptoms of inflammation, namely redness (rubor), swelling (tumour), heat (calor), pain (dolor) and loss of function (functio laesa), are only part of the story. The term inflammation is taken to include the full range of underlying cellular and molecular mechanisms involved, not only in the production of the inflammatory responses but, more importantly in clinical terms, in the healing process as well. Thus the journal covers molecular, cellular, animal and clinical studies, and related aspects of pharmacology, such as anti-inflammatory drug development, trials and therapeutic developments. It also considers publication of negative findings. Journal of Inflammation aims to become the leading online journal on inflammation and, as online journals replace printed ones over the next decade, the main open access inflammation journal. Open access guarantees a larger audience, and thus impact, than any restricted access equivalent, and increasingly so, as the escalating costs of printed journals puts them outside University budgets. The unrestricted access to research findings in inflammation aids in promoting dynamic and productive dialogue between industrial and academic members of the inflammation research community, which plays such an important part in the development of future generations of anti-inflammatory therapies.