Effect of spleen on the ability of hepatic macrophages to activate hepatic stellate cells in the progression of liver fibrosis

Shaoying Zhang; Dan Wan; Xi Deng; Xiao Liang; Fanfan Liang; Chongyu Zhang; Jiazhen Zhu; Yang Zhao; Zongfang LI

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Effect of spleen on the ability of hepatic macrophages to activate hepatic stellate cells in the progression of liver fibrosis

VernacularTitle:肝纤维化进展中脾脏对肝脏巨噬细胞活化肝星状细胞作用的影响
Author: Shaoying ZHANG ¹ ; Dan WAN ¹ ; Xi DENG ² ; Xiao LIANG ¹ ; Fanfan LIANG ¹ ; Chongyu ZHANG ¹ ; Jiazhen ZHU ³ ; Yang ZHAO ¹ ; Zongfang LI ^{1
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Author Information

1. National & Local Joint Engineering Research Center of Biodiagnosis & Biotherapy, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004, China
2. Department of Geriatric General Surgery, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004, China
3. Shaanxi Provincial Clinical Medical Research Center for Liver and Spleen Diseases, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004, China
4. Shaanxi Provincial Clinical Medical Research Center for Liver and Spleen Diseases, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004, China
5. Department of Geriatric General Surgery, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710004, China
Publication Type:Journal Article
Keywords: spleen; hepatic macrophage; inflammatory cytokine; hepatic stellate cell; glutamine metabolism
From: Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(4):575-581
CountryChina
Language:Chinese
Abstract: 【Objective】 To investigate the effect of spleen on hepatic macrophages mediated activation of hepatic stellate cells (HSCs) in mice with liver fibrosis. 【Methods】 Eighteen male C57BL/6 mice were randomly divided into three groups. Mice in Group A and Group B were injected intraperitoneally with CCl4 to establish liver fibrosis mouse model, while those in Group C were injected with corn oil as normal control. Four weeks later, mice with liver fibrosis received splenectomy (Spx) or sham operation (Sham), respectively. After continuous injection for 2 weeks, liver homogenates (L-Homo) were prepared and liver cells were isolated from the three groups. Expressions of IL-1β, IL-13, TGF-β, TNF-α, PDGF-β and VEGF in the liver homogenates of the three groups were detected by Luminex multifactor analysis. The expressions of these cytokines in liver macrophages (L-Mφ) and other non-parenchymal cells of Sham and Spx mice were analyzed by Real-time quantitative PCR (RT-qPCR) and flow cytometry. Macrophage cell line RAW264.7 or bone marrow-derived macrophages (BMDMs) were treated with liver homogenates from the Sham and Spx groups. Then the differently treated RAW264.7 cells were analyzed for mRNA expressions of cytokines and glutamine metabolism-related molecules by RT-qPCR, or transwell co-cultured with hepatic stellate cell line JS1. After co-culture, the survival and extracellular matrix expression of JS1 cells were analyzed. For comparison, Student’s t test (between two groups) or one-way analysis of variance (among multiple groups) were used. 【Results】 Compared with normal control group, the concentrations of IL-1β, IL-13, TGF-β and TNF-α in the L-Homo of model group were significantly increased and showed higher levels in Sham group than in Spx group. Moreover, the hepatic macrophages were indicated as the major source of these cytokines. Consistently, macrophages treated with liver homogenate of Sham mice had increased expressions of IL-1β, TGF-β and TNF-α and glutaminase (GLS). After co-culture with macrophages treated with liver homogenate of Sham group rather than Spx group, JS1 expressed higher expressions of α-SMA and collagens. 【Conclusion】 The spleen is involved in regulating the secretion of cytokines by hepatic macrophages and enhancing their ability to activate hepatic stellate cells.