Proteomic Changes of Activated Hepatic Stellate Cells

Hepatic stellate cells (HSC) are the major cellular drivers of liver fibrosis. Upon liver inflammation caused by a broad range of insults including non-alcoholic fatty liver, HSC transform from a quiescent into a proliferating, fibrotic phenotype. Although much is known about the pathophysiology of this process, exact cellular processes which occur in HSC and enable this transformation remain yet to be elucidated. In order to investigate this HSC transformation, we employed a simple, yet reliable model of HSC activation via an increase in growth medium serum concentration (serum activation). For that purpose, immortalized human LX-2 HSC were exposed to either 1% or 10% fetal bovine serum (FBS). Resulting quiescent (1% FBS) and activated (10% FBS) LX-2 cells were then subjected to in-depth mass spectrometry-based proteomics analysis as well as comprehensive phenotyping. Protein network analysis of activated LX-2 cells revealed an increase in the production of ribosomal proteins and proteins related to cell cycle control and migration, resulting in higher proliferation and faster migration phenotypes. Interestingly, we also observed a decrease in the expression of cholesterol and fatty acid biosynthesis proteins in accordance with a concomitant loss of cytosolic lipid droplets during activation. Overall, this work provides an update on HSC activation characteristics using contemporary proteomic and bioinformatic analyses and presents an accessible model for HSC activation. Data are available via ProteomeXchange with identifier PXD029121.

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Network Pharmacological Analysis and Experimental Validation of the Mechanisms of Action of Si-Ni-San Against Liver Fibrosis

Frontiers in Pharmacology ◽

10.3389/fphar.2021.656115 ◽

2021 ◽

Vol 12 ◽

Author(s):

Siliang Wang ◽

Cheng Tang ◽

Heng Zhao ◽

Peiliang Shen ◽

Chao Lin ◽

...

Keyword(s):

Liver Fibrosis ◽

Hepatic Stellate Cells ◽

Stellate Cells ◽

Mechanisms Of Action ◽

Network Pharmacology ◽

Related Factor ◽

Alcoholic Fatty Liver ◽

Ppar Γ ◽

Liver Tissues

Background: Si-Ni-San (SNS), a commonly used traditional Chinese medicine (TCM) formula, has potency against liver diseases, such as hepatitis and non-alcoholic fatty liver disease (NAFLD). However, the therapeutic efficacy and pharmacological mechanisms of action of SNS against liver fibrosis remain largely unclear.Methods: A carbon tetrachloride (CCl4)-induced liver fibrosis mouse model was adopted for the first time to investigate the beneficial effects of SNS on liver fibrosis. The potential mechanisms of action of SNS were explored using the network pharmacology-based strategy and validated with the aid of diverse assays.Results: SNS treatment reduced collagen and ECM deposition, downregulated fibrosis-related factor (hyaluronic acid and laminin) contents in serum, maintained the morphological structure of liver tissue, and improved liver function in the liver fibrosis model. Based on network pharmacology results, apoptosis, inflammation and angiogenesis, together with the associated pathways (including VEGF, TNF, caspase, PPAR-γ and NF-κB), were identified as the mechanisms underlying the effects of SNS on liver fibrosis. Further in vivo experiments validated the significant mitigatory effects of SNS on inflammatory infiltration and pro-inflammatory cytokine contents (IFNγ, IL-1β and TGF-β1) in liver tissues of mice with liver fibrosis. SNS suppressed pathologic neovascularization as well as levels of VEGFR1, VEGF and VEGFR2 in liver tissues. SNS treatment additionally inhibited hepatic parenchyma cell apoptosis in liver tissues of mice with liver fibrosis and regulated apoptin expression while protecting L02 cells against apoptosis induced by TNF-α and Act D in vitro. Activation of hepatic stellate cells was suppressed and the balance between MMP13 and TIMP1 maintained in vitro by SNS. These activities may be associated with SNS-induced NF-κB suppression and PPAR-γ activation.Conclusion: SNS effectively impedes liver fibrosis progression through alleviating inflammation, ECM accumulation, aberrant angiogenesis and apoptosis of hepatic parenchymal cells along with inhibiting activation of hepatic stellate cells through effects on multiple targets and may thus serve as a novel therapeutic regimen for this condition.

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Extension of Long Cellular Processes of Hepatic Stellate Cells Cultured on Extracellular Type I Collagen Gel by Microtubule Assembly: Observation Utilizing Time-lapse Video-microscopy.

Cell Structure and Function ◽

10.1247/csf.22.487 ◽

1997 ◽

Vol 22 (4) ◽

pp. 487-492 ◽

Cited By ~ 13

Author(s):

Mitsutaka Miura ◽

Mitsuru Sato ◽

Itaru Toyoshima ◽

Haruki Senoo

Keyword(s):

Hepatic Stellate Cells ◽

Type I Collagen ◽

Collagen Gel ◽

Stellate Cells ◽

Time Lapse ◽

Microtubule Assembly ◽

Type I ◽

Cellular Processes ◽

Type I Collagen Gel ◽

Time Lapse Video

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Nitrovasodilators inhibit platelet-derived growth factor-induced proliferation and migration of activated human hepatic stellate cells

Gastroenterology ◽

10.1053/gast.2000.9354 ◽

2000 ◽

Vol 119 (2) ◽

pp. 479-492 ◽

Cited By ~ 80

Author(s):

Paola Failli ◽

Raffaella M.S. DeFranco ◽

Alessandra Caligiuri ◽

Alessandra Gentilini ◽

Roberto Giulio Romanelli ◽

...

Keyword(s):

Growth Factor ◽

Hepatic Stellate Cells ◽

Stellate Cells ◽

Platelet Derived Growth Factor ◽

And Migration ◽

Proliferation And Migration

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Metabolic Hallmarks of Hepatic Stellate Cells in Liver Fibrosis

Cells ◽

10.3390/cells9010024 ◽

2019 ◽

Vol 9 (1) ◽

pp. 24 ◽

Cited By ~ 7

Author(s):

Olga Khomich ◽

Alexander V. Ivanov ◽

Birke Bartosch

Keyword(s):

Liver Disease ◽

Liver Fibrosis ◽

Hepatic Stellate Cells ◽

Current Knowledge ◽

Stellate Cells ◽

Central Carbon Metabolism ◽

Regenerative Process ◽

Therapeutic Interventions ◽

Alcoholic Fatty Liver ◽

Liver Functions

Liver fibrosis is a regenerative process that occurs after injury. It is characterized by the deposition of connective tissue by specialized fibroblasts and concomitant proliferative responses. Chronic damage that stimulates fibrogenic processes in the long-term may result in the deposition of excess matrix tissue and impairment of liver functions. End-stage fibrosis is referred to as cirrhosis and predisposes strongly to the loss of liver functions (decompensation) and hepatocellular carcinoma. Liver fibrosis is a pathology common to a number of different chronic liver diseases, including alcoholic liver disease, non-alcoholic fatty liver disease, and viral hepatitis. The predominant cell type responsible for fibrogenesis is hepatic stellate cells (HSCs). In response to inflammatory stimuli or hepatocyte death, HSCs undergo trans-differentiation to myofibroblast-like cells. Recent evidence shows that metabolic alterations in HSCs are important for the trans-differentiation process and thus offer new possibilities for therapeutic interventions. The aim of this review is to summarize current knowledge of the metabolic changes that occur during HSC activation with a particular focus on the retinol and lipid metabolism, the central carbon metabolism, and associated redox or stress-related signaling pathways.

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MiR-9a-5p regulates proliferation and migration of hepatic stellate cells under pressure through inhibition of Sirt1

World Journal of Gastroenterology ◽

10.3748/wjg.v21.i34.9900 ◽

2015 ◽

Vol 21 (34) ◽

pp. 9900 ◽

Cited By ~ 17

Author(s):

Feng Qi

Keyword(s):

Hepatic Stellate Cells ◽

Stellate Cells ◽

And Migration ◽

Proliferation And Migration

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GJA1 promotes hepatocellular carcinoma progression by mediating TGF-β-induced activation and the epithelial–mesenchymal transition of hepatic stellate cells

Open Medicine ◽

10.1515/med-2021-0344 ◽

2021 ◽

Vol 16 (1) ◽

pp. 1459-1471

Author(s):

Gengming Niu ◽

Xiaotian Zhang ◽

Runqi Hong ◽

Ximin Yang ◽

Jiawei Gu ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Conditioned Medium ◽

Hepatic Stellate Cells ◽

Hepatic Cirrhosis ◽

Epithelial Mesenchymal Transition ◽

Stellate Cells ◽

Gene Set Enrichment Analysis ◽

Mesenchymal Transition ◽

And Migration ◽

Proliferation And Migration

Abstract Introduction Gap junction protein, alpha 1 (GJA1), which is correlated with recurrences and unfavorable prognoses in hepatocellular carcinomas (HCCs), is one of the specific proteins expressed by activated hepatic stellate cells (HSCs). Methods Expression of GJA1 was compared between HCCs and nontumor tissues (NTs), between hepatic cirrhosis and NTs, and between primary and metastatic HCCs using transcriptomic datasets from the Gene Expression Omnibus and the Integrative Molecular Database of Hepatocellular Carcinoma. The in vitro activities of GJA1 were investigated in cultured HSCs and HCC cells. The underlying mechanism was characterized using Gene Set Enrichment Analysis and validated by western blotting. Results The expression of GJA1 was significantly increased in HCCs and hepatic cirrhosis compared to that in NTs. GJA1 was also overexpressed in pulmonary metastases from HCCs when compared with HCCs without metastasis. Overexpression of GJA1 promoted while knockdown of GJA1 inhibited proliferation and transforming growth factor (TGF)-β-mediated activation and migration of cultured HSCs. Overexpression of GJA1 by lentivirus infection promoted proliferation and migration, while conditioned medium from HSCs overexpressing GJA1 promoted migration but inhibited proliferation of Hep3B and PLC-PRF-5 cells. Lentivirus infection with shGJA1 or conditioned medium from shGJA1-infected HSCs inhibited the proliferation and migration of HCCLM3 cells that had a high propensity toward lung metastasis. Mechanistically, GJA1 induced the epithelial–mesenchymal transition (EMT) in HSCs and HCCLM3 cells. Conclusion GJA1 promoted HCC progression by inducing HSC activation and the EMT in HSCs. GJA1 is potentially regulated by TGF-β and thus may be a therapeutic target to inhibit HCC progression.

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MiR-571 affects the development and progression of liver fibrosis by regulating the Notch3 pathway

10.21203/rs.3.rs-360390/v1 ◽

2021 ◽

Author(s):

Shuo Cong ◽

Yongmei Liu ◽

Yi Li ◽

Yu Chen ◽

Rui Chen ◽

...

Keyword(s):

Liver Fibrosis ◽

Western Blot ◽

Signaling Pathway ◽

Hepatic Stellate Cells ◽

Stellate Cells ◽

Rt Pcr ◽

Promote Cell Proliferation ◽

Notch 3 ◽

And Migration ◽

Set Up

Abstract Exploring the expression of miR-571 in patients with liver fibrosis and its role in the progression of liver fibrosis. A total of 74 patients with chronic hepatitis and cirrhosis accompanied by liver fibrosis in our institution from September to December 2018 were collected for study, and the expression of miR-571 in patients with different progressions of liver fibrosis was determined by RT-PCR and Western blot analysis. Set up Notch3 up group and Notch3 down regulated group, RT-PCR and Western blot were used to determine the effect of Notch signaling on the expression of fibrogenic α-SMA, collagen I. CCK-8, cell scratch assays, Transwell assays, flow cytometry were used to determine the effect of miR-571 on LX-2 proliferation, migration, apoptosis in human stem stellate cells, and RT-PCR, Western blot assays were performed to determine the effect of miR-571 on the Notch3 signaling pathway and the expression of profibrogenic factors. miR-571 is up-regulated in patients with liver fibrosis and is associated with the progression of liver fibrosis. Notch3 signaling pathway can promote the expression of fibroblast in human hepatic stellate cells; miR-571 can inhibit the apoptosis of human hepatic stellate cells, promote cell proliferation and migration; up regulation of miR-571 can promote the expression of Notch3 and Jagged 1; up regulation of miR-571 can also promote the expression of fibroblast. miR-571 can promote the activation of human stem stellate cells and the expression of fibroblasts through Notch 3 signaling pathway.

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