BET选择性抑制剂38号化合物对急性肝损伤小鼠保护作用研究*

doi:10.3969/j.issn.1672-5069.2022.05.004

摘要/Abstract

摘要： 目的探讨溴域和末端外结构域(BET)选择性抑制剂38号化合物对CCl₄诱导的小鼠急性肝损伤(ALI)的保护作用及其机制。方法使用脂多糖(LPS)刺激Raw264.7巨噬细胞,诱导急性细胞炎症模型,设对照组、LPS处理组和LPS与38号化合物共培养组,提取细胞RNA,采用RT-qPCR法检测炎症相关细胞因子mRNA水平。构建CCl₄诱导的ALI模型,将24只小鼠随机分为对照组、模型组和药物干预组。采用免疫组化法检测肝组织抗F4/80 和抗Ly-6G阳性细胞。结果 LPS刺激细胞模型组IL-1βmRNA水平为(23246.0±1185.0),显著高于对照组【(1.1±0.1),P＜0.05】,细胞IL-6 mRNA水平为(7740.0±322.2),显著高于对照组【(1.1±0.4),P＜0.05】,TNF-αmRNA水平为(132.2±2.7),显著高于对照组【(1.0±0),P＜0.05】,而38号化合物干预处理后细胞IL-1β、IL-6和TNF-αmRNA水平均显著降低,在80 nM处理组分别为(2409.0±147.6)、(66.0±2.1)和(36.7±1),在40 nM处理组分别为(4790.0±206.4)、(314.1±10.7)和(47.0±1.5),在20 nM处理组分别为(10079.0±500.3)、(1112.0±22.0)和(73.0±1.8),在10 nM处理组分别为[(13794.0±1025.0)、(2576.0±46.3)和(96.8±7.2),P＜0.05],呈现浓度依赖性;ALI小鼠血清ALT和AST水平分别为(8281.0±2710.0)U/L和(5330.0±2435.0)U/L,显著高于对照组[分别为(51.5±7.0)U/L和(215.3±12.4)U/L,P＜0.05],而干预组分别为(3634.0±713.9.0) U/L和(2876.0±667.1)U/L,较模型组显著降低(P＜0.05);ALI小鼠模型肝组织结构紊乱,炎症反应明显,肝细胞大片坏死,大量的炎症细胞聚集,而药物干预组上述病理学变化均较模型组减轻。结论 BET选择性抑制剂38号化合物能减轻由CCl₄诱导的小鼠急性肝损伤,改善肝功能和肝组织结构的破坏,对小鼠肝损伤具有保护作用,其机制可能与抑制了炎症相关的细胞因子表达,从而减少了炎症细胞的聚集有关。

关键词: 急性肝损伤, 溴域和末端外结构域抑制剂, 38号化合物, Raw264.7巨噬细胞, 小鼠

Abstract: Objective The aim of this study was to investigate the protective effect of compound 38, a selective bromodomain and extra-terminal (BET) inhibitor, on CCl₄-induced acute liver injury (ALI) in mice. Methods The Raw264.7 cells was stimulated by LPS with or without compound 38 co-culture, and the Raw264.7 cells with no stimulation served as control. The cell RNA was extracted by Trizol, and the cell inflammation-related genes were detected by RT-qPCR. Twenty-four mice were divided randomly into control, model and compound 38-intervened group, and the model was established by CCl₄administration intraperitoneally. Serum ALT and AST level was measured, and the pathological changes of liver tissues were observed. The intrahepatic neutrophil and macrophage was detected by immunohistochemistry. Results In model cells, the IL-1βmRNA, IL-6 mRNA and TNF-αmRNA levels were (23246.0±1185.0), (7740.0±322.2) and (132.2±2.7), significantly higher than [(1.1±0.1), (1.1±0.4) and (1.0±0), P＜0.05], while in the compound 38-intervened cells were all greatly dose-dependently decreased as compared to those in the model (all P＜0.05); serum ALT and AST levels in mice with ALI were (8281.0±2710.0)U/L and (5330.0±2435.0)U/L, significantly higher than [(51.5±7.0)U/L) and (215.3±12.4)U/L, respectively, P＜0.05] in the control, while in the compound 38-intervended cells were (3634.0±713.9.0) U/L and (2876.0±667.1)U/L, greatly decreased compared to in the model (P＜0.05); the histopathological examination showed that the liver tissue structure was disordered with obvious inflammatory response, such as hepatocyte necrosis, destruction of normal lobule structure and aggregation of a large number of inflammatory cells in the model group, while the above pathological changes were alleviated in the intervention group. Conclusion The copound 38, a selective BET inhibitor, could alleviate CCl₄-induced acute liver injury in mice, and the mechanism might be related to the inhibition of inflammation-related cytokine expression and decreased infiltration of macrophages.

Key words: Acute liver injures, Bromodomain and extra-terminal inhibitor, Compound 38, Raw264.7 macrophages, Mice

付荣, 吴康卉, 石翠翠, 范建高, 李光明. BET选择性抑制剂38号化合物对急性肝损伤小鼠保护作用研究^*[J]. 实用肝脏病杂志, 2022, 25(5): 620-623.

Fu Rong, Wu Kanghui, Shi Cuicui, et al.. Protective effect of BET selective inhibitor, compound 38 on acute liver injury in mice[J]. Journal of Practical Hepatology, 2022, 25(5): 620-623.

参考文献

[1]Kim S J, Lee S M. NLRP3 inflammasome activation in D-galactosamine and lipopolysaccharide-induced acute liver failure: role of heme oxygenase-1. Free Radic Biol Med, 2013, 65:997-1004.
[2]Filippakopoulos P, Knapp S. Targeting bromodomains: epigenetic readers of lysine acetylation. Nat Rev Drug Discov, 2014, 13(5): 337-356.
[3]Arrowsmith C H, Bountra C, Fish P V, et al. Epigenetic protein families: a new frontier for drug discovery. Nat Rev Drug Discov, 2012, 11(5): 384-400.
[4]Shi J, Vakoc C R. The mechanisms behind the therapeutic activity of BET bromodomain inhibition. Mol Cell, 2014, 54(5): 728-736.
[5]Filippakopoulos P, Picaud S, Mangos M, et al. Histone recognition and large-scale structural analysis of the human bromodomain family. Cell, 2012, 149(1): 214-231.
[6]Borck P C, Guo L-W, Plutzky J. BET epigenetic reader proteins in cardiovascular transcriptional programs. Circ Res, 2020, 126(9): 1190-1208.
[7]Duan Q, Wu P, Liu Z, et al. BET bromodomain inhibition suppresses adipogenesis in mice. Endocrine, 2020, 67(1): 264-267.
[8]Kulikowski E, Rakai B D, Wong N C W. Inhibitors of bromodomain and extra-terminal proteins for treating multiple human diseases. Med Res Rev, 2021, 41(1): 223-245.
[9]Shu S, Lin C Y, He H H, et al. Response and resistance to BET bromodomain inhibitors in triple-negative breast cancer. Nature, 2016, 529(7586): 413-417.
[10] Farzanegi P, Dana A, Ebrahimpoor Z, et al. Mechanisms of beneficial effects of exercise training on non-alcoholic fatty liver disease (NAFLD): Roles of oxidative stress and inflammation. Eur J Sport Sci, 2019, 19(7):994-1003.
[11] Matyas C, Haskó G, Liaudet L, et al. Interplay of cardiovascular mediators, oxidative stress and inflammation in liver disease and its complications. Nat Rev Cardiol, 2021, 18(2): 117-135.
[12] Mansouri A, Gattolliat C H, Asselah T. Mitochondrial dysfunction and signaling in chronic liver diseases. Gastroenterology, 2018, 155(3): 629-647.
[13] Li Z, Xiao S, Yang Y, et al. Discovery of 8-methyl-pyrrolo[1,2-]pyrazin-1(2)-one derivatives as highly potent and selective bromodomain and extra-terminal (BET) bromodomain inhibitors. J Med Chem, 2020, 63(8): 3956-3975.
[14] Dai C, Xiao X, Li D, et al. Chloroquine ameliorates carbon tetrachloride-induced acute liver injury in mice via the concomitant inhibition of inflammation and induction of apoptosis. Cell Death Dis, 2018, 9(12): 1-13.
[15] Peng J, Li J, Huang J, et al. p300/CBP inhibitor A-485 alleviates acute liver injury by regulating macrophage activation and polarization. Theranostics, 2019, 9(26): 8344-8361.
[16] Liu Y, Li J, Liao L, et al. Cyclin-dependent kinase inhibitor roscovitine attenuates liver inflammation and fibrosis by influencing initiating steps of liver injury. Clin Sci (Lond), 2021, 135(7): 925-941.
[17] Mukhopadhyay P, Rajesh M, Cao Z, et al. Poly (ADP-ribose) polymerase-1 is a key mediator of liver inflammation and fibrosis. Hepatology, 2014, 59(5): 1998-2009.
[18] Doherty D G. Immunity, tolerance and autoimmunity in the liver: A comprehensive review. J Autoimmun, 2016, 66: 60-75.
[19] Heymann F, Tacke F. Immunology in the liver-from homeostasis to disease. Nat Rev Gastroenterol Hepatol, 2016, 13(2):88-110.
[20] Robinson M W, Harmon C, O'farrelly C. Liver immunology and its role in inflammation and homeostasis. Cell Mol Immunol, 2016, 13(3): 267-276.
[21] Macpherson A J, Heikenwalder M, Ganal-Vonarburg S C. The liver at the nexus of host-microbial interactions. Cell Host Microbe, 2016, 20(5): 561-571.
[22] Bai P, Ye H, Xie M, et al. A synthetic biology-based device prevents liver injury in mice. J Hepatol, 2016, 65(1): 84-94.

BET选择性抑制剂38号化合物对急性肝损伤小鼠保护作用研究^*

Protective effect of BET selective inhibitor, compound 38 on acute liver injury in mice

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