scholarly journals TWIST2 and the PPAR signaling pathway are important in the progression of nonalcoholic steatohepatitis

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Yanmei Zhang ◽  
Xiaoxiao Ge ◽  
Yongqing Li ◽  
Bingyang Zhang ◽  
Peijun Wang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Fatty Liver ◽  
Rna Sequencing ◽  
Nonalcoholic Steatohepatitis ◽  
In Vitro Model ◽  
Bhlh Transcription Factor ◽  
Protein Levels ◽  
Ppar Signaling ◽  
Vitro Model

Abstract Background To investigate the roles of the transcription factors twist family bHLH transcription factor 1 (TWIST1), twist family bHLH transcription factor 2 (TWIST2), and peroxisome proliferator activated receptor gamma (PPARγ) in the progression of nonalcoholic steatohepatitis. Methods The protein levels of TWIST1, TWIST2 and PPARγ were determined in the serum of nonalcoholic fatty liver disease (NAFLD) patients and healthy controls by enzyme-linked immunosorbent assay (ELISA). An in vivo model for fatty liver was established by feeding C57BL/6 J mice a high-fat diet (HFD). An in vitro model of steatosis was established by treating LO-2 cells with oleic acid (OA). RNA sequencing was performed on untreated and OA-treated LO-2 cells followed by TWIST1, TWIST2 and PPARγ gene mRNA levels analysis, Gene Ontology (GO) enrichment and pathway analysis. Results The TWIST2 serum protein levels decreased significantly in all fatty liver groups (P < 0.05), while TWIST1 varied. TWIST2 tended to be lower in mice fed an HFD and was significantly lower at 3 months. Similarly, in the in vitro model, the TWIST2 protein level was downregulated significantly at 48 and 72 h after OA treatment. RNA sequencing of LO-2 cells showed an approximately 2.3-fold decrease in TWIST2, with no obvious change in TWIST1 and PPARγ. The PPAR signaling pathway was enriched, with 4 genes upregulated in OA-treated cells (P = 0.0018). The interleukin (IL)-17 and tumor necrosis factor (TNF) signaling pathways were enriched in OA-treated cells. Conclusions The results provide evidence that the TWIST2 and PPAR signaling pathways are important in NAFLD and shed light on a potential mechanism of steatosis.

scholarly journals Investigating the Role of Transcription Factors TWIST1, TWIST2 and PPARγ in the Progression of Nonalcoholic Steatohepatitis

2020 ◽  
Author(s):  
Yanmei Zhang ◽  
Xiaoxiao Ge ◽  
Yongqing Li ◽  
Sumei Lu ◽  
Wanshan Ma
Keyword(s):  
Transcription Factors ◽  
Fatty Liver ◽  
Rna Sequencing ◽  
Nonalcoholic Steatohepatitis ◽  
Mrna Levels ◽  
Protein Levels ◽  
Ppar Signaling ◽  
Vitro Model

Abstract Background: To investigate the role that transcription factors TWIST1, TWIST2 and PPARγ play in nonalcoholic steatohepatitis progression. Methods: The protein levels of TWIST1, TWIST2 and PPARγ were determined in the serum of NAFLD patients and healthy controls by ELISA. An in vivo model for fatty liver was established by feeding C57BL/6J mice a high fat diet (HFD). An in vitro model of steatosis was established by treating LO-2 cells with oleic acid (OA). RNA sequencing was performed on the untreated and OA treated LO-2 cells followed by TWIST1, TWIST2 and PPARγ gene mRNA levels analysis, Gene Ontology (GO) enrichment and pathway analysis. Results: The TWIST2 serum protein levels decreased significantly in all fatty liver groups (P<0.05) while TWIST1 varied. TWIST2 tended to be lower in mice fed a HFD and was significantly lower at 3 months. Similarly, in our in vitro model, TWIST2 protein level was down significantly at 48 and 72 hours after OA treatment. RNA sequencing of the LO-2 cells showed an approximately 2.3-fold decrease in TWIST2, with TWIST1 and PPARγ no obvious change. The PPAR signaling pathway was enriched with 4 genes up regulated in OA treated cells (P=0.0018). IL-17 and TNF signaling pathways were enriched in OA treated cells. Conclusions: Our results provide evidence that TWIST2 and PPARγ are important in NAFLD and shed light on a potential mechanism of steatosis.

scholarly journals Effects of tacrolimus on autophagy protein LC3 in puromycin-damaged mouse podocytes

2020 ◽  
Vol 48 (12) ◽  
pp. 030006052097142
Author(s):  
Xiao-qing Yang ◽  
Sheng-you Yu ◽  
Li Yu ◽  
Lin Ge ◽  
Yao Zhang ◽  
...  
Keyword(s):  
Fluorescence Intensity ◽  
Light Chain ◽  
In Vitro Model ◽  
Group Versus ◽  
Podocyte Injury ◽  
Apoptosis Rate ◽  
Protein Levels ◽  
Vitro Model ◽  
Intercellular Connections

Objective To investigate the mechanism through which tacrolimus, often used to treat refractory nephropathy, protects against puromycin-induced podocyte injury. Methods An in vitro model of puromycin-induced podocyte injury was established by dividing podocytes into three groups: controls, puromycin only (PAN group), and puromycin plus tacrolimus (FK506 group). Podocyte morphology, number, apoptosis rate and microtubule associated protein 1 light chain 3 alpha ( LC3) expression were compared. Results Puromycin caused podocyte cell body shrinkage and loose intercellular connections, but podocyte morphology in the FK506 group was similar to controls. The apoptosis rate was lower in the FK506 group versus PAN group. The low level of LC3 mRNA observed in untreated podocytes was decreased by puromycin treatment; however, levels of LC3 mRNA were higher in the FK506 group versus PAN group. Although LC3-I and LC3-II protein levels were decreased by puromycin, levels in the FK506 group were higher than the PAN group. Fewer podocyte autophagosomes were observed in the control and FK506 groups versus the PAN group. Cytoplasmic LC3-related fluorescence intensity was stronger in control and FK506 podocytes versus the PAN group. Conclusions Tacrolimus inhibited puromycin-induced mouse podocyte damage by regulating LC3 expression and enhancing autophagy.

scholarly journals Induction of Immunoglobulin Heavy-Chain Transcription through the Transcription Factor Bright Requires TFII-I

2006 ◽  
Vol 26 (12) ◽  
pp. 4758-4768 ◽  
Author(s):  
Jaya Rajaiya ◽  
Jamee C. Nixon ◽  
Neil Ayers ◽  
Zana P. Desgranges ◽  
Ananda L. Roy ◽  
...  
Keyword(s):  
Transcription Factor ◽  
B Lymphocytes ◽  
Heavy Chain ◽  
In Vitro Model ◽  
Model System ◽  
Interaction Domain ◽  
Tyrosine Residues ◽  
Immunoglobulin Locus ◽  
Vitro Model

ABSTRACT Bright/ARID3a/Dril1, a member of the ARID family of transcription factors, is expressed in a highly regulated fashion in B lymphocytes, where it enhances immunoglobulin transcription three- to sixfold. Recent publications from our lab indicated that functional, but not kinase-inactive, Bruton's tyrosine kinase (Btk) is critical for Bright activity in an in vitro model system, yet Bright itself is not appreciably tyrosine phosphorylated. These data suggested that a third protein, and Btk substrate, must contribute to Bright-enhanced immunoglobulin transcription. The ubiquitously expressed transcription factor TFII-I was identified as a substrate for Btk several years ago. In this work, we show that TFII-I directly interacts with human Bright through amino acids in Bright's protein interaction domain and that specific tyrosine residues of TFII-I are essential for Bright-induced activity of an immunoglobulin reporter gene. Moreover, inhibition of TFII-I function in a B-cell line resulted in decreased heavy-chain transcript levels. These data suggest that Bright functions as a three-component protein complex in the immunoglobulin locus and tie together previous data indicating important roles for Btk and TFII-I in B lymphocytes.

1250 HUMAN IN VITRO MODEL OF FATTY LIVER AND THE INFLUENCE OF ANTI-LIPIDEMIC DRUGS ON METABOLISM AND SIGNAL PATHWAYS IN STEATOTIC HEPATOCYTES

2013 ◽  
Vol 58 ◽  
pp. S506-S507
Author(s):  
G. Damm ◽  
A. Krüger ◽  
S. Lünse ◽  
S. Sperling ◽  
V. Kegel ◽  
...  
Keyword(s):  
Fatty Liver ◽  
In Vitro Model ◽  
Signal Pathways ◽  
Vitro Model

scholarly journals Three-dimensional perfused human in vitro model of non-alcoholic fatty liver disease

2017 ◽  
Vol 23 (2) ◽  
pp. 204 ◽  
Author(s):  
Tomasz Kostrzewski ◽  
Terri Cornforth ◽  
Sophie A Snow ◽  
Larissa Ouro-Gnao ◽  
Cliff Rowe ◽  
...  
Keyword(s):  
Liver Disease ◽  
Fatty Liver ◽  
Fatty Liver Disease ◽  
In Vitro Model ◽  
Three Dimensional ◽  
Alcoholic Fatty Liver ◽  
Vitro Model ◽  
Alcoholic Fatty Liver Disease

scholarly journals SUN-550 CARF Through Its Lipid Lowering Effect May Play a Pivotal Role in the Development of Non-Alcoholic Fatty Liver Diseases

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Kamrul M Hasan ◽  
Meher Parveen ◽  
Alondra Pena ◽  
Amiya P Sinha-Hikim ◽  
Theodore C Friedman
Keyword(s):  
Lipid Metabolism ◽  
Fatty Liver ◽  
Hepg2 Cells ◽  
In Vitro Model ◽  
Lipid Lowering ◽  
Alcoholic Fatty Liver ◽  
Lowering Effect ◽  
Vitro Model ◽  
Lipid Lowering Effect

Abstract CARF (Collaborator of ARF), a member of ARF-MDM2-p53 pathway and an emerging multifunctional protein, regulates cellular fate in response to various stresses including oxidative DNA damage and replicative stresses. However, its role in metabolic syndrome (MS) and associated diseases has not been studied. This study, using our well established in vivo and in vitro model systems, examines the role of CARF in the development of non-alcoholic fatty liver disease (NAFLD). Indeed, we have found that, compared to control, CARF expression along with Sirt1, pAMPK and pACC (common biological markers of NAFLD) was significantly decreased in the nicotine and high-fat-diet (HFD) in combination or HFD alone induced fatty livers. Additionally, CARF expression was down regulated in palmitate (PA)-treated HepG2 cells, an in vitro model of steatosis, suggesting that CARF expression is negatively regulated in MS, such as NAFLD. Our study further revealed that shRNA mediated knockdown or lentiviral mediated over expression of CARF induced or reduced endogenous fat accumulation, respectively, in HepG2 cells. We also found that overexpression of CARF lowered the exogenous fat accumulation in PA treated HepG2 cells. RNA seq analysis after CARF knockdown in HEK-293T cells further revealed that genes associated with lipid metabolism and triglyceride (TG) synthesis such as diacylglycerol O-acyltransferase2 (DGAT2), acyl-CoA synthetase long-chain family member 4 and 6 (ACSL4, ACSL6) were upregulated in CARF-depleted cells. Likewise, we also found increased expression of DGAT2 in CARF-depleted HepG2 cells, which enhanced TG synthesis. Intriguingly, consistent with the lipid lowering effects of metformin, an antidiabetic drug, we further found that CARF expression along with pAMPK and Sirt1 were significantly increased in metformin-treated HepG2 cells. However, we also found increased pACC levels in CARF over-expressing cells which was further enhanced in metformin-treated cells, suggesting, for the first time, that CARF may contribute to lipid lowering effect of metformin by inhibiting lipogenesis. We conclude that CARF has a lipid lowering effect in hepatocytes and its down regulation in response to MS perturbs lipid metabolism that may lead to the development of NAFLD.

scholarly journals High-Fat Diet-Induced Fatty Liver Is Associated with Immunosuppressive Response during Sepsis in Mice

2021 ◽  
Vol 2021 ◽  
pp. 1-18
Author(s):  
Fangzhao Wang ◽  
Zhongran Cen ◽  
Zhanguo Liu ◽  
Jianwei Gan ◽  
Xianglong Zhang ◽  
...  
Keyword(s):  
Fatty Liver ◽  
High Fat Diet ◽  
In Vitro Model ◽  
High Fat ◽  
Metabolic Disturbances ◽  
Differential Signaling ◽  
Normal Chow ◽  
Severe Liver Damage ◽  
Vitro Model

High-fat diet-induced fatty liver is an indolent and chronic disease accompanied by immune dysfunction and metabolic disturbances involving numerous biological pathways. This study investigated how this abnormal metabolic disorder influences sepsis in mice. Mice were fed with normal chow (NC) or high-fat diet (HFD), and palmitic acid (PA) was used to treat hepatocytes to mimic fat accumulation in vitro. Lipopolysaccharide (LPS) was used to induce sepsis and related immune responses. Mice fed on a high-fat diet displayed higher mortality and more severe liver damage but compromised immunoreaction. The supernatant from PA-treated primary hepatocytes markedly diminished the inflammatory cytokine expression of macrophages after LPS stimulation, which showed a state of immunosuppression. Metabolomics analysis indicated the level of many key metabolites with possible roles in immunoreaction was altered in the HFD and PA groups compared with corresponding controls; specifically, β-hydroxybutyric acid (BHB) showed an immunosuppressive effect on Raw264.7 cells during the LPS stimulation. Transcriptomic analysis suggested that several differential signaling pathways may be associated with the alteration of immune function between the NC and HFD groups, as well as in the in vitro model. Our study suggests that the consumption of HFD may alter the hepatic metabolic profile, and that certain metabolites may remold the immune system to immunosuppressive state in the context of sepsis.

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