Full length articleCD73 regulates hepatic stellate cells activation and proliferation through Wnt/β-catenin signaling pathway
Graphical abstract
Introduction
Alcoholic liver fibrosis (ALF), a sustained scarring response, caused by long-term alcohol consumption, is characterized by the accumulation of excessive amounts of extracellular matrix (ECM). Persisting liver fibrogenesis may progress to liver cirrhosis or hepatocellular carcinoma (Liu et al., 2019; Xu et al., 2019). Hepatic stellate cells (HSCs) activation, the critical event of hepatic fibrogenesis, leads to the abundant synthesis and secretion of ECM and exerts an important role in the up-regulation of α-smooth muscle actin (α-SMA) (Zhang et al., 2018). Given the important role of HSCs in liver fibrogenesis, promoting the apoptosis of activated HSCs is a critical step in fibrosis reversion (Ding et al., 2019; Peverill et al., 2014). Accumulating evidence illustrates that the development of ALF in alcoholics is mainly due to the metabolism of ethanol to the highly reactive compound acetaldehyde (Mello et al., 2008; Wang et al., 2015). Acetaldehyde, acting on HSCs in a paracrine manner, can directly increase the expression of Col1α1 and deposition of ECM by activating multiple signaling pathways and transcription factors (Gao and Bataller, 2011; Setshedi et al., 2010). Recent years have yielded a wealth of data implicating that the activation of HSCs stimulated with acetaldehyde in vitro closely mimics the activation of HSCs in vivo and ultimately results in ALF (Wang et al., 2014; Yang et al., 2016).
Ecto-5′-nucleotidase (CD73), a membrane-bound nucleotide that breakdowns extracellular ATP into adenosine, has emerged as an important regulator of pathophysiologic responses related to inflammation, tissue fibrosis, and cancer (Beavis et al., 2012; Buchheiser et al., 2011). Previous studies have identified high expression of CD73 in a multitude of cancer cell types including hepatocellular carcinoma and the favorable role of tumor growth and metastasis (Ma et al., 2019; Zhang, 2010, 2014). Furthermore, blocking CD73 can effectively alleviate suppression of the adaptive immune system and inhibit the growth and metastasis of tumors (Koszałka et al., 2015). In prior work, we found that blockade of adenosine A1 and A2A receptors prevented the development of ALF, indicating that extracellular adenosine and its receptors play a vital role in the pathophysiology of ALF with complex mechanisms (Wang et al., 2020; Yang et al., 2015). Moreover, CD73 catalyzes the terminal step of ATP formation to extracellular adenosine and functions in modulating adenosine signaling (Zimmermann et al., 2012). Therefore, it is meaningful for the study to explore the role of CD73 in the progression of ALF.
Numerous reports have confirmed that Wnt/β-catenin pathway is a pivotal regulator of cell growth and proliferation and is activated in many human organ fibrosis (Guo et al., 2012; Nishikawa et al., 2018). Other studies have demonstrated that Wnt/β-catenin pathway is closely related to the proliferation of HSCs and liver fibrosis (Yu et al., 2019). Mechanically, previous studies identified that CD73 could be involved in adenosine metabolism via interacting with Wnt/β-catenin pathway (Spychala, 2000; Spychala and Kitajewski, 2004). Therefore, whether CD73 regulates the proliferation and activation of HSCs through Wnt/β-catenin pathway has captured the attention of our team.
Section snippets
Mouse model of ALF
C57BL/6 J mice (male, 8–10 weeks of age, 21–24 g) were purchased from the Animal Experiment Center of Anhui Medical University. All experiments were conducted as required of the Ethics Committee and Animal Experimental Committee at Anhui Medical University. We replicated the mouse model of ALF based on the study by B. Gao et al. (Kwon et al., 2014) and used APCP, an inhibitor of CD73, to knockdown CD73 expression in vivo. The mice were randomly divided into six groups of 12 animals: vehicle
Upregulation of CD73 expression in HSCs may be associated with ethanol plus CCl4-induced liver fibrosis
To define whether CD73 expression is associated with ALF, ethanol plus CCl4-induced mouse liver fibrosis model was developed for histopathological examination. First, Liver injury in liver fibrosis mice was evaluated by H&E staining. Masson and Sirius red staining were used to indicate the collagen area, and the activated HSCs were highlighted by α-SMA positivity (Fig. 1A). Moreover, Serum levels of AST and ALT were higher (P < 0.01) in ethanol plus CCl4-induced liver fibrosis mice compared
Discussion
With the striking increase in alcohol consumption, ALF has been widely recognized as a vital global health issue and the research on its pathogenesis and therapeutic targets has become the hotspot of global focus (Bala and Szabo, 2012; Huang et al., 2018). Fortunately, ALF is a reversible process that strikes a balance between the synthesis and degradation of ECM that leads to abnormal deposition of fibrous connective tissue in response to various liver injuries (Huang et al., 2020). Activated
Data availability
All data included in this study are available upon request by contact with the corresponding author.
CRediT authorship contribution statement
Wen-qian Jia: designed the research and wrote the main manuscript text. Tao-cheng Zhou: directed . Jing-wen Dai: directed . Zhen-ni Liu: directed . Ya-fei Zhang: directed the data analysis, All authors read and approved the manuscript. Dan-dan Zang: designed the research and wrote the main manuscript text, directed the data analysis, All authors read and approved the manuscript.
Declaration of competing interest
The authors declare no conflicts of interest.
Acknowledgments
This study was supported by the National Natural Science Foundation of China (Grant No. 81270498 and Grant No. 81970518). The authors thank the Center for Scientific Research of Anhui Medical University for valuable help in our experiment.
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