scholarly journals Silybin Modulates Collagen Turnover in an In Vitro Model of NASH

Molecules ◽  
2019 ◽  
Vol 24 (7) ◽  
pp. 1280 ◽  
Author(s):  
Beatrice Anfuso ◽  
Pablo Giraudi ◽  
Claudio Tiribelli ◽  
Natalia Rosso
Keyword(s):  
Cell Viability ◽  
In Vitro Model ◽  
Early Stage ◽  
Antioxidant Properties ◽  
Ros Generation ◽  
Driving Mechanism ◽  
Beneficial Effects ◽  
Huh7 Cells ◽  
Vitro Model

Silybin has been proposed as a treatment for nonalcoholic steatohepatitis (NASH). In this study, we assessed the effect of Silybin in a well-established in vitro coculture model of early-stage NASH. LX2 and Huh7 cells were exposed to free fatty acid (FFA) and Silybin as mono- or coculture (SCC). Cell viability, LX2 activation, collagen deposition, metalloproteinase 2 and 9 (MMP2-9) activity, and ROS generation were determined at 24, 96, and 144 h. Exposure to FFA induced the activation of LX2 as shown by the increase in cell viability and upregulation of collagen biosynthesis. Interestingly, while cotreatment with Silybin did not affect collagen production in LX2, a significant reduction was observed in SCC. MMP2-9 activity was reduced in FFA-treated Huh7 and SCC and cotreatment with Silybin induced a dose-dependent increase, while no effect was observed in LX2. Silybin also showed antioxidant properties by reducing the FFA-induced production of ROS in all the cell systems. Based on these data, Silybin exerts its beneficial effects by reducing LX2 proliferation and ROS generation. Moreover, MMP2-9 modulation in hepatocytes represents the driving mechanism for the net reduction of collagen in this NASH in vitro model, highlighting the importance of hepatic cells interplay in NASH development and resolution.

scholarly journals Establishment of an In Vitro Model of Persistent Chicken Anemia Virus Infection

Pathogens ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 842
Author(s):  
Hieu Van Dong ◽  
Giang Thi Huong Tran ◽  
Dai Quang Trinh ◽  
Yohei Takeda ◽  
Haruko Ogawa ◽  
...  
Keyword(s):  
Cell Viability ◽  
Persistent Infection ◽  
In Vitro Model ◽  
Neutralizing Antibody ◽  
Chicken Anemia Virus ◽  
Viral Gene ◽  
Vitro Model ◽  
Anemia Virus ◽  
Cells Cultured

Persistent infection of chicken anemia virus (CAV) in chickens has been suspected to result in immunosuppression and exogenous virus contamination within vaccine production. However, no direct evidence for persistent CAV infection has thus far been obtained. In this study, we aimed to establish an in vitro model of persistent CAV infection. CAV-infected MDCC-MSB1 (MSB1) cells, a Marek’s disease virus-transformed continuous cell line, were cultured in the presence of both CAV and CAV neutralizing antibody (NA). Cell viability, expression of viral antigens, viral DNA, and recovery of CAV were examined by acridine orange/propidium iodide staining, immunofluorescence measurement, real-time PCR, and viral isolation, respectively. The results indicated that CAV was maintained and possibly replicated in CAV-infected cells cultured in the presence of NA, without affecting host cell viability. It was also shown that persistently infectious CAV induced cell death again after removing NA. The persistent infection of CAV in MSB1 cells was not related to viral gene mutation. In summary, we have herein established a novel model of persistent CAV infection in MSB1 cells cultured in the presence of NA.

Photobiomodulation increases cell viability via AKT activation in an in vitro model of diabetes induced by glucose neurotoxicity

2019 ◽  
Vol 35 (1) ◽  
pp. 149-156 ◽  
Author(s):  
Victória Regina da Silva Oliveira ◽  
Rosangela Aparecida Santos-Eichler ◽  
Camila Squarzoni Dale
Keyword(s):  
Cell Viability ◽  
In Vitro Model ◽  
Akt Activation ◽  
Vitro Model

Development of an in vitro model of the early-stage bovine tuberculous granuloma using Mycobacterium bovis-BCG

2015 ◽  
Vol 168 (3-4) ◽  
pp. 249-257 ◽  
Author(s):  
Laura Garza-Cuartero ◽  
Elaine McCarthy ◽  
Joseph Brady ◽  
Joseph Cassidy ◽  
Clare Hamilton ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
In Vitro Model ◽  
Early Stage ◽  
Mycobacterium Bovis Bcg ◽  
Tuberculous Granuloma ◽  
Vitro Model

scholarly journals Hypoxia induces stress fiber formation in adipocytes in the early stage of obesity

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Golnaz Anvari ◽  
Evangelia Bellas
Keyword(s):  
In Vitro Model ◽  
Nuclear Translocation ◽  
Early Stage ◽  
Coiled Coil ◽  
Stress Fiber ◽  
Fiber Formation ◽  
Actin Stress Fiber ◽  
Stress Fiber Formation ◽  
Vitro Model

AbstractIn obese adipose tissue (AT), hypertrophic expansion of adipocytes is not matched by new vessel formation, leading to AT hypoxia. As a result, hypoxia inducible factor-1⍺ (HIF-1⍺) accumulates in adipocytes inducing a transcriptional program that upregulates profibrotic genes and biosynthetic enzymes such as lysyl oxidase (LOX) synthesis. This excess synthesis and crosslinking of extracellular matrix (ECM) components cause AT fibrosis. Although fibrosis is a hallmark of obese AT, the role of fibroblasts, cells known to regulate fibrosis in other fibrosis-prone tissues, is not well studied. Here we have developed an in vitro model of AT to study adipocyte-fibroblast crosstalk in a hypoxic environment. Further, this in vitro model was used to investigate the effect of hypoxia on adipocyte mechanical properties via ras homolog gene family member A (RhoA)/Rho-associated coiled-coil kinases (ROCK) signaling pathways. We confirmed that hypoxia creates a diseased phenotype by inhibiting adipocyte maturation and inducing actin stress fiber formation facilitated by myocardin-related transcription factor A (MRTF-A/MKL1) nuclear translocation. This work presents new potential therapeutic targets for obesity by improving adipocyte maturation and limiting mechanical stress in obese AT.

Beneficial effects of curcumin on GFAP filament organization and down-regulation of GFAP expression in an in vitro model of Alexander disease

2012 ◽  
Vol 318 (15) ◽  
pp. 1844-1854 ◽  
Author(s):  
Tiziana Bachetti ◽  
Eleonora Di Zanni ◽  
Pietro Balbi ◽  
Roberto Ravazzolo ◽  
GianPietro Sechi ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Alexander Disease ◽  
Down Regulation ◽  
Beneficial Effects ◽  
Gfap Expression ◽  
Vitro Model

scholarly journals A novel methylated analogue of L-Mimosine exerts its therapeutic potency through ROS production and ceramide-induced apoptosis in malignant melanoma

2021 ◽  
Author(s):  
Sotiris Kyriakou ◽  
William Cheung ◽  
Theodora Mantso ◽  
Melina Mitsiogianni ◽  
Ioannis Anestopoulos ◽  
...  
Keyword(s):  
Malignant Melanoma ◽  
In Vitro Model ◽  
Ros Generation ◽  
Intrinsic Apoptosis ◽  
Membrane Depolarisation ◽  
Vitro Model ◽  
Induced Apoptosis ◽  
Related Proteins ◽  
Metabolomic Approach

SummaryMelanoma is an aggressive and highly metastatic type of skin cancer where the design of new therapies is of utmost importance for the clinical management of the disease. Thus, we have aimed to investigate the mode of action by which a novel methylated analogue of L-Mimosine (e.g., L-SK-4) exerts its therapeutic potency in an in vitro model of malignant melanoma. Cytotoxicity was assessed by the Alamar Blue assay, oxidative stress by commercially available kits, ROS generation, caspase 3/7 activation and mitochondrial membrane depolarisation by flow cytometry, expression of apoptosis-related proteins by western immunoblotting and profiling of lipid biosynthesis by a metabolomic approach. Overall, higher levels of ROS, sphingolipids and apoptosis were induced by L-SK-4 suggesting that the compound’s therapeutic potency is mediated through elevated ROS levels which promote the upregulation of sphingolipid (ceramide) biosynthesis thus leading to the activation of both extrinsic and intrinsic apoptosis, in an experimental model of malignant melanoma.

scholarly journals Trehalose Modulates Autophagy Process to Counteract Gliadin Cytotoxicity in an In Vitro Celiac Disease Model

Cells ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 348 ◽  
Author(s):  
Federico Manai ◽  
Alberto Azzalin ◽  
Martina Morandi ◽  
Veronica Riccardi ◽  
Lisa Zanoletti ◽  
...  
Keyword(s):  
Celiac Disease ◽  
In Vitro Model ◽  
Wheat Gluten ◽  
Disease Model ◽  
Autoimmune Disorder ◽  
Beneficial Effects ◽  
Gliadin Peptides ◽  
Autophagy Pathway ◽  
Vitro Model

Celiac disease (CD) is a chronic systemic autoimmune disorder that is triggered by the ingestion of gliadin peptides, the alcohol-soluble fraction of wheat gluten. These peptides, which play a key role in the immune response that underlies CD, spontaneously form aggregates and exert a direct toxic action on cells due to the increase in the reactive oxygen species (ROS) levels. Furthermore, peptic-tryptic digested gliadin peptides (PT-gliadin) lead to an impairment in the autophagy pathway in an in vitro model based on Caco-2 cells. Considering these premises, in this study we have analyzed different mTOR-independent inducers, reporting that the disaccharide trehalose, a mTOR-independent autophagy activator, rescued the autophagy flux in Caco-2 cells treated with digested gliadin, as well as improved cell viability. Moreover, trehalose administration to Caco-2 cells in presence of digested gliadin reduced the intracellular levels of these toxic peptides. Altogether, these results showed the beneficial effects of trehalose in a CD in vitro model as well as underlining autophagy as a molecular pathway whose modulation might be promising in counteracting PT-gliadin cytotoxicity.

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