Hexa Histidine-Tagged Recombinant Human Cytoglobin Deactivates Hepatic Stellate Cells and Inhibits Liver Fibrosis by Scavenging Reactive Oxygen Species.
Hexa Histidine-Tagged Recombinant Human Cytoglobin Deactivates Hepatic Stellate Cells and Inhibits Liver Fibrosis by Scavenging Reactive Oxygen Species.
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六组氨酸标记的重组人细胞球蛋白通过清除活性氧使肝星状细胞失活并抑制肝纤维化。
DOI：
10.1002/hep.31752
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发表时间：
2021-06
期刊：
影响因子：
--
通讯作者：
Kawada N
中科院分区：
文献类型：
--
作者：
Dat NQ;Thuy LTT;Hieu VN;Hai H;Hoang DV;Thi Thanh Hai N;Thuy TTV;Komiya T;Rombouts K;Dong MP;Hanh NV;Hoang TH;Sato-Matsubara M;Daikoku A;Kadono C;Oikawa D;Yoshizato K;Tokunaga F;Pinzani M;Kawada N
抗纤维化治疗仍然是人类慢性肝病中未满足的医疗需求。我们报道了细胞珠蛋白（CYGB）的抗纤维化特性，CYGB是一种在肝星状细胞（HSC）中表达的呼吸蛋白，HSC是参与肝纤维化的主要细胞类型。 患有胆管结扎诱导的肝胆汁淤积或胆碱缺乏的氨基酸限定饮食诱导的脂肪性肝炎的Cygb缺陷小鼠显著加重了肝损伤、纤维化和活性氧（ROS）形成。所有这些表现在Cygb过表达小鼠中减弱。我们生产了六组氨酸标记的重组人CYGB（His‐CYGB），追踪其生物分布，并使用硫代乙酰胺（TAA）或3，5-二乙氧羰基-1，4-二氢可力丁（DDC）评估其在HSC或晚期肝硬化小鼠中的功能。在培养的HSC中，细胞外的His‐CYGB通过网格蛋白介导的途径被内吞并积聚在核内体中。His-CYGB显著阻碍HSC中ROS自发形成或在存在ROS诱导剂的情况下形成，从而导致1型胶原α 1产生和α-平滑肌肌动蛋白表达减弱。用钴替代血红素组的铁中心使His‐CYGB的作用无效。此外，His-CYGB诱导HSC分泌干扰素-β，这部分有助于其抗纤维化功能。Momelotinib不完全逆转His‐CYGB的作用。在给予TAA或DDC的小鼠中，静脉注射His‐CYGB显著抑制肝脏炎症、纤维化和氧化性细胞损伤，而无不良反应。RNA测序分析揭示了细胞培养物和肝组织中炎症和纤维化相关基因的下调以及抗氧化基因的上调。注射的His-CYGB主要定位于HSC，但不定位于巨噬细胞，表明特异性靶向作用。His-CYGB在具有人源化肝脏的嵌合小鼠中未表现出毒性。His-CYGB可能对人类慢性肝病具有抗纤维化临床应用。
Antifibrotic therapy remains an unmet medical need in human chronic liver disease. We report the antifibrotic properties of cytoglobin (CYGB), a respiratory protein expressed in hepatic stellate cells (HSCs), the main cell type involved in liver fibrosis. Cygb‐deficient mice that had bile duct ligation–induced liver cholestasis or choline‐deficient amino acid–defined diet–induced steatohepatitis significantly exacerbated liver damage, fibrosis, and reactive oxygen species (ROS) formation. All of these manifestations were attenuated in Cygb‐overexpressing mice. We produced hexa histidine–tagged recombinant human CYGB (His‐CYGB), traced its biodistribution, and assessed its function in HSCs or in mice with advanced liver cirrhosis using thioacetamide (TAA) or 3,5‐diethoxycarbonyl‐1,4‐dihydrocollidine (DDC). In cultured HSCs, extracellular His‐CYGB was endocytosed and accumulated in endosomes through a clathrin‐mediated pathway. His‐CYGB significantly impeded ROS formation spontaneously or in the presence of ROS inducers in HSCs, thus leading to the attenuation of collagen type 1 alpha 1 production and α‐smooth muscle actin expression. Replacement the iron center of the heme group with cobalt nullified the effect of His‐CYGB. In addition, His‐CYGB induced interferon‐β secretion by HSCs that partly contributed to its antifibrotic function. Momelotinib incompletely reversed the effect of His‐CYGB. Intravenously injected His‐CYGB markedly suppressed liver inflammation, fibrosis, and oxidative cell damage in mice administered TAA or DDC mice without adverse effects. RNA‐sequencing analysis revealed the down‐regulation of inflammation‐ and fibrosis‐related genes and the up‐regulation of antioxidant genes in both cell culture and liver tissues. The injected His‐CYGB predominantly localized to HSCs but not to macrophages, suggesting specific targeting effects. His‐CYGB exhibited no toxicity in chimeric mice with humanized livers. His‐CYGB could have antifibrotic clinical applications for human chronic liver diseases.
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影响因子：
82.9
作者：
Friedman SL;Neuschwander-Tetri BA;Rinella M;Sanyal AJ
通讯作者：
Sanyal AJ
影响因子：
4.6
作者：
Thuy le TT;Van Thuy TT;Matsumoto Y;Hai H;Ikura Y;Yoshizato K;Kawada N
通讯作者：
Kawada N
影响因子：
7.6
作者：
Hasegawa, T;Yoneda, M;Terano, A
通讯作者：
Terano, A
影响因子：
3.2
作者：
ANDERSON, WA
通讯作者：
ANDERSON, WA
影响因子：
29.4
作者：
GUALDI, R;CASALGRANDI, G;PIETRANGELO, A
通讯作者：
PIETRANGELO, A