抑制糖原合成酶激酶3β（GSK3β）的活性可通过抑制多种程序性细胞死亡来减轻急性肝衰竭。
Inhibition of GSK3β activity alleviates acute liver failure via suppressing multiple programmed cell death.
作者信息
Zhang Danmei, Shi Chunxia, Zhang Qingqi, Wang Yukun, Guo Jin, Gong Zuojiong
机构信息
Department of Infectious Diseases, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuhan, Hubei province, 430060, China.
出版信息
J Inflamm (Lond). 2023 Jul 13;20(1):24. doi: 10.1186/s12950-023-00350-1.
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.
背景
急性肝衰竭（ALF）是成人中最常见的危及生命的疾病之一，患者既往无肝病。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并促进自噬，从而发挥保护作用。在本研究中，我们旨在探讨抑制GSK3β后对急性肝衰竭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）在体外和体内诱导急性肝衰竭。然后使用GSK3β抑制剂TDZD-8来探究其对急性肝衰竭的保护作用。TDZD-8处理后，分别采用TUNEL染色和流式技术检测肝组织和细胞中的凋亡比例，同时进行蛋白质免疫印迹法和免疫荧光测定以检测组织和细胞中凋亡、焦亡和坏死性凋亡相关蛋白的表达水平。此外，进行蛋白质免疫印迹法以探究GSK3β抑制后肝保护作用的具体机制，单独抑制TRAF6和HDAC3后检测TAK1、TRAF6和HDAC3的表达水平。通过免疫荧光检测TRAF6和HDAC3在体外的共定位，同时通过免疫沉淀试验检测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均可显著缓解急性肝衰竭的进展。抑制GSK3β活性可显著降低肝细胞凋亡、焦亡、坏死性凋亡水平，并改善肝功能障碍和组织损伤。此外，我们发现急性肝衰竭时肝细胞TAK1和TRAF6水平降低，HDAC3水平升高，而抑制GSK3β可上调TAK1和TRAF6水平并降低HDAC3表达。
结论
GSK3β抑制剂TDZD-8可预防急性肝衰竭的进展，其作用可能涉及TRAF6/HDAC3/TAK1通路。
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