title: A20 Attenuates Liver Fibrosis in NAFLD and Inhibits Inflammation Responses.
abstract: We previously reported A20 was able to inhibit lipid accumulation in nonalcoholic steatohepatitis. We want to investigate whether A20 influences liver fibrosis in this study. Liver tissues from patients with hepatic fibrosis (n = 9) and healthy individuals (n = 7) were studied for A20 protein level by immunohistochemistry. A20 messenger RNA (mRNA) and protein level were also analyzed in two murine hepatic fibrosis models: methionine- and choline-deficient (MCD) diet and extrahepatic bile duct ligation (BDL) operation by real-time PCR and western blot. In vitro, the LX-2 human hepatic stellate cell line was treated by LPS at 0, 0.001, 0.01, 0.1, and 1 μg/mL for 6 h or at the concentration of 0.1 μg/mL for 0, 6, 12, and 24 h, then A20 expression levels were detected by western blot and PCR. The mRNA level of α-SMA, collagen I, collagen III, TGF-β, IL-6, MCP-1, and TLR4 was also examined by PCR. We then overexpressed A20 in LX-2 cells using adenovirus technique. Levels of α-SMA, collagen I, collagen III, TGF-β, IL-6, MCP-1, and TLR4 were examined in A20-overexpression LX-2 cells. Patients with hepatic fibrosis showed significantly higher A20 protein level compared with healthy controls. A20 mRNA and protein levels were also increased in livers from MCD feeding or BDL operation mice in comparison to normal controls. In LX-2 cells, LPS induced A20 protein in a concentration-dependent manner. The mRNA levels of α-SMA, collagen I, collagen III, TGF-β, IL-6, MCP-1, and TLR4 were increased after LPS treatment. Overexpression of A20 in LX-2 cells inhibited α-SMA deposition and collagen I, collagen III secretion. TGF-β, IL-6, MCP-1, and TLR4 mRNA levels were also reduced in A20-overexpression LX-2 cells in response to LPS stimulation. A20 overexpression inhibits hepatic stellate cell activation, which could be the mechanism for high A20 expression protected livers from fibrosis. Enhancement of A20 expression seems to be rational therapeutic strategies for liver fibrosis.
authors.affiliation: Department of Health Manage Center, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, No.160, Pujian Road, Shanghai, 200127, China.|Department of Gastroenterology and Hepatology, The First Hospital of Jiaxing, 1882 Central-South Road, Jiaxing, Zhejiang Province, China.
authors.forename: Xiaohan
authors.identifier: http://orcid.org/0000-0002-2779-5566
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Health Manage Center, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, No.160, Pujian Road, Shanghai, 200127, China.|Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Digestive Disease, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Luoyan
authors.initials: L
authors.lastname: Ai
authors.affiliation: Department of Health Manage Center, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, No.160, Pujian Road, Shanghai, 200127, China.|Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Digestive Disease, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Xu
authors.affiliation: Department of Health Manage Center, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, No.160, Pujian Road, Shanghai, 200127, China.|Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Digestive Disease, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Changwei
authors.initials: C
authors.lastname: Wu
authors.affiliation: Department of Health Manage Center, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, No.160, Pujian Road, Shanghai, 200127, China.|Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Digestive Disease, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Health Manage Center, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, No.160, Pujian Road, Shanghai, 200127, China.|Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Digestive Disease, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Dazhi
authors.initials: D
authors.lastname: Su
authors.affiliation: Department of Health Manage Center, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, No.160, Pujian Road, Shanghai, 200127, China.|Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Institute of Digestive Disease, 145 Middle Shandong Road, Shanghai, 200001, China.
authors.forename: Xiaoke
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Department of Health Manage Center, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, No.160, Pujian Road, Shanghai, 200127, China. zhuping_fan@163.com.
authors.forename: Zhuping
authors.initials: Z
authors.lastname: Fan
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D016022:Case-Control Studies; D024042:Collagen Type I / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D055166:Hepatic Stellate Cells; D006801:Humans; D007249:Inflammation / Q000517:prevention & control*; D008070:Lipopolysaccharides; D008103:Liver Cirrhosis / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D065626:Non-alcoholic Fatty Liver Disease / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3 / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D024042:Collagen Type I; D016207:Cytokines; D008070:Lipopolysaccharides; D012333:RNA, Messenger; C544013:alpha-smooth muscle actin, mouse; C076529:TNFAIP3 protein, human; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3; C513261:Tnfaip3 protein, mouse
keywords: A20; LX-2 cells; inflammation; liver fibrosis