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Alvo em GPR65 alivia a inflamação hepática e a fibrose suprimindo as vias JNK e NF-κB

  • role： First author第一作者
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

  • Introduction：Kun Zhang, Meng-Xia Zhang and Xiao-Xiang Meng contributed equally to this work
Kun Zhang1,  
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

  • Introduction：Kun Zhang, Meng-Xia Zhang and Xiao-Xiang Meng contributed equally to this work
Meng-Xia Zhang1,  
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

  • Introduction：Kun Zhang, Meng-Xia Zhang and Xiao-Xiang Meng contributed equally to this work
Xiao-Xiang Meng1,  
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

Jing Zhu1,  
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

Jia-Jun Wang1,  
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

Yi-Fan He1,  
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

Ye-Hua Li1,  
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

Si-Cong Zhao1,  
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

Zhe-Min Shi1,  
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

Li-Na Zheng1,  
  • role： Corresponding author通信作者
  • Affiliation：

    Department of Hepatology and Gastroenterology, Tianjin Union Medical Center, Tianjin Medical University, Tianjin Union Medical Center Affiliated to Nankai University, Tianjin 300000, China

  • Email：hantaomd@126.com
  • Introduction：*hantaomd@126.com;
Tao Han2*,  
  • role： Corresponding author通信作者
  • Affiliation：

    Department of Histology and Embryology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

  • Email：hongwei@tmu.edu.cn
  • Introduction：hongwei@tmu.edu.cn
Wei Hong1*

Resumo

Background:G-protein coupled receptors (GPCRs) are recognized as attractive targets for drug therapy. However, it remains poorly understood how GPCRs, except for a few chemokine receptors, regulate the progression of liver fibrosis. Here, we aimed to reveal the role of GPR65, a proton-sensing receptor, in liver fibrosis and to elucidate the underlying mechanism.Methods:The expression level of GPR65 was evaluated in both human and mouse fibrotic livers. Furthermore, Gpr65-deficient mice were treated with either bile duct ligation (BDL) for 21 d or carbon tetrachloride (CCl4) for 8 weeks to investigate the role of GPR65 in liver fibrosis. A combination of experimental approaches, including Western blotting, quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR), and enzyme-linked immunosorbent assay (ELISA), confocal microscopy and rescue studies, were used to explore the underlying mechanisms of GPR65’s action in liver fibrosis. Additionally, the therapeutic potential of GPR65 inhibitor in the development of liver fibrosis was investigated.Results:We found that hepatic macrophage (HM)-enriched GPR65 was upregulated in both human and mouse fibrotic livers. Moreover, knockout of Gpr65 significantly alleviated BDL- and CCl4-induced liver inflammation, injury and fibrosis in vivo, and mouse bone marrow transplantation (BMT) experiments further demonstrated that the protective effect of Gpr65 knockout is primarily mediated by bone marrow-derived macrophages (BMMs). Additionally, in vitro data demonstrated that Gpr65 silencing and GPR65 antagonist inhibited, while GPR65 overexpression and application of GPR65 endogenous and exogenous agonists enhanced the expression and release of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and transforming growth factor-β (TGF-β), all of which subsequently promoted the activation of hepatic stellate cells (HSCs) and the damage of hepatocytes (HCs). Mechanistically, GPR65 overexpression, the acidic pH and GPR65 exogenous agonist induced up-regulation of TNF-α and IL-6 via the Gαq-Ca2+-JNK/NF-κB pathways, while promoted the expression of TGF-β through the Gαq-Ca2+-MLK3-MKK7-JNK pathway. Notably, pharmacological GPR65 inhibition retarded the development of inflammation, HCs injury and fibrosis in vivo.Conclusions:GPR65 is a major regulator that modulates the progression of liver fibrosis. Thus, targeting GPR65 could be an effective therapeutic strategy for the prevention of liver fibrosis.

Palavras-chave

GPR65;Hepatic fibrosis;Hepatic macrophages;Inflammation;c-Jun N-terminal kinase;Nuclear factor κB

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