Cellular and molecular effects of silymarin on the transdifferentiation processes of LX-2 cells and its connection with lipid metabolism

Silva, Caio Mateus da; Ferrari, Gustavo Duarte; Alberici, Luciane C.; Malaspina, Osmar; Moraes, Karen C. M.

doi:10.1007/s11010-020-03717-7

Cited by 9 publications

(6 citation statements)

References 100 publications

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“…The results of DMSO application were found to be dependent on substrate stiffness; DMSO significantly ( Figure 5 B) or insignificantly ( Figure 7 B) decreased the collagen content on the soft gel, but had no clear effects on the stiff gel ( Figure 5 D). The effects of DMSO on collagen gene expression or this protein content have been reported previously [ 36 , 37 ].…”

Section: Discussionmentioning

confidence: 65%

The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases

Gałdyszyńska

Radwańska

Szymański

et al. 2021

Cells

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Information about mechanical strain in the extracellular space is conducted along collagen fibers connected with integrins and then transmitted within cells. An aim of the study is to verify the hypothesis that the stiffness of cardiac human fibroblast substrates exerts a regulatory effect on collagen metabolism via integrin α2β1 and downstream signaling. The experiments were performed on human cardiac fibroblasts cultured on stiff or soft polyacrylamide gels. Extracellular and intracellular collagen content, metalloproteinase-1 (MMP-1), metalloproteinase-9 (MMP-9) and expression of the α1 chain of the procollagen type I gene (Col1A1) were elevated in cultures settled on soft substrate. The substrate stiffness did not modify tissue inhibitors of matrix metalloproteinase capacity (TIMPs 1–4). Integrin α2β1 inhibition (TC-I 15) or α2 subunit silencing resulted in augmentation of collagen content within the culture. Expression of Col1A1 and Col3A1 genes was increased in TC-I 15-treated fibroblasts. Total and phosphorylated levels of both FAK and Src kinases were elevated in fibroblasts cultured on stiff substrate. Inhibition of FAK (FAK kinase inhibitor 14) or Src kinase (AZM 47527) increased collagen content within the culture. The substrate stiffness exerted a regulatory influence on collagen metabolism via integrin α2β1 and its downstream signaling (FAK and Src kinases) in cardiac fibroblasts.

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Section: Discussionmentioning

confidence: 65%

The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases

Gałdyszyńska

Radwańska

Szymański

et al. 2021

Cells

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“…Even though serum activation of human LX-2 cells has been used a number of times before [ 18 , 19 , 32 , 33 , 34 , 35 ] it has not yet been characterized in depth with regard to changes in gene and protein expression compared to rat HSC serum activation [ 36 ]. Therefore, we first confirmed that the addition of FBS to growth media is both an easy and reliable way to activate LX-2 cells.…”

Section: Discussionmentioning

confidence: 99%

Proteomic Changes of Activated Hepatic Stellate Cells

Schinagl

Tomin

Gindlhuber

et al. 2021

IJMS

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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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“…Fluorescent microscopy analyses for fatty acid tracking inside the cells were performed according to Rambold et al [ 33 ] and Silva et al [ 34 ] For the assays, 3 × 10 4 cells were cultivated on circular coverslips in 24‐well plates, and cells were treated as previously described (see Section 2.2). To the culture medium containing or not fipronil, 1 µM of BODIPY™ 558/568 C 12 (Bodipy‐C 12 ) (4,4‐difluoro‐5‐(2‐thienyl)‐4‐bora‐3a,4a‐diaza‐s‐indacene‐3‐dodecanoic acid) (Thermo Fisher Scientific) was added to different groups of cells.…”

Section: Methodsmentioning

confidence: 99%

“…It was performed through the LD labeling with 200 ng/mL of BODIPY™ 493/503 (4,4‐difluoro‐1,3,5,7,8‐pentamethyl‐4‐Bora‐3a,4a‐diaza‐sIndacene) (Thermo Fisher Scientific) for 20 min, followed by consecutive washes in PBS. Cell nuclei were stained with DAPI following the protocol described in Da Silva et al [ 34 ] The images were taken as previously described (see Section 2.4) and quantified using the Image‐J software (http://rsbweb.nih.gov) with the “JACoP” plug‐in [ 35 ] to obtain a Pearson's coefficient between Bodipy‐C 12 overlap and lipid droplets.…”

Section: Methodsmentioning

confidence: 99%

“…The effects of fipronil on β-oxidation of fatty acid (FA) in HepG2 cells were investigated using microscopy analyses of the pulse-chase method to track BODIPY-C12. [33,34] The analyses investigated the cellular capacity to maintain C-12 in lipid droplets inside the cells in different cultures during the investigated time. The cells exposed to the concentration of 25 μM of etomoxir were set as the experimental control of the assay.…”

Section: Effects Of Fipronil On β-Oxidation and Glucose Metabolism In...mentioning

confidence: 99%

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Cellular and molecular effects of fipronil in lipid metabolism of HepG2 and its possible connection to non‐alcoholic fatty liver disease

Pansa,

Molica,

de Oliveira Júnior

et al. 2023

J Biochem & Molecular Tox

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Nonalcoholic fatty liver disease (NAFLD) is a global public health problem that affects more than a quarter of the population. The development of this disease is correlated with metabolic dysfunctions that lead to lipid accumulation in the liver. Pesticides are one of etiologies that support NAFLD establishment. Therefore, the effects of the insecticide fipronil on the lipid metabolism of the human hepatic cell line, HepG2, was investigated, considering its widespread use in field crops and even to control domestic pests. To address the goals of the study, biochemical, cellular, and molecular analyses of different concentrations of fipronil in cell cultures were investigated, after 24 h of incubation. Relevant metabolites such as triglycerides, glucose levels, β‐oxidation processes, and gene expression of relevant elements correlated with lipid and metabolism of xenobiotics were investigated. The results suggested that at 20 μM, the pesticide increased the accumulation of triglycerides and neutral lipids by reducing fatty acid oxidation and increasing de novo lipogenesis. In addition, changes were observed in genes that control oxidative stress and the xenobiotic metabolism. Together, the results suggest that the metabolic changes caused by the insecticide fipronil may be deleterious if persistent, favoring the establishment of hepatic steatosis.

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Cellular and molecular effects of silymarin on the transdifferentiation processes of LX-2 cells and its connection with lipid metabolism

Cited by 9 publications

References 100 publications

The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases

The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases

Proteomic Changes of Activated Hepatic Stellate Cells

Cellular and molecular effects of fipronil in lipid metabolism of HepG2 and its possible connection to non‐alcoholic fatty liver disease

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