Various strategies have been employed to improve the reliability of 2D, 3D, and co-culture in vitro models of nonalcoholic fatty liver disease, including using extracellular matrix proteins such as collagen I to promote cell adhesion. While studies have demonstrated the significant benefits of culturing cells on collagen I, its effects on the HepG2 cell line after exposure to palmitate (PA) have not been investigated. Therefore, this study aimed to assess the effects of PA-induced lipotoxicity in HepG2 cultured in the absence or presence of collagen I. HepG2 cultured in the absence or presence of collagen I was exposed to PA, followed by analyses that assessed cell proliferation, viability, adhesion, cell death, mitochondrial respiration, reactive oxygen species production, gene and protein expression, and triacylglycerol accumulation. Culturing HepG2 on collagen I was associated with increased cell proliferation, adhesion, and expression of integrin receptors, and improved cellular spreading compared to culturing them in the absence of collagen I. However, PA-induced lipotoxicity was greater in collagen I-cultured HepG2 than in those cultured in the absence of collagen I and was associated with increased α2β1 receptors. In summary, the present study demonstrated for the first time that collagen I-cultured HepG2 exhibited exacerbated cell death following exposure to PA through integrin-mediated death. The findings from this study may serve as a caution to those using 2D models or 3D scaffold-based models of HepG2 in the presence of collagen I.

• MeSH
• buněčná adheze * účinky léků   MeSH
• buněčná smrt účinky léků   MeSH
• buňky Hep G2 MeSH
• integrin alfa2beta1 metabolismus   MeSH
• integriny metabolismus   genetika   MeSH
• kolagen typu I * metabolismus   genetika   MeSH
• lidé MeSH
• nealkoholová steatóza jater metabolismus   patologie   MeSH
• palmitany toxicita   farmakologie   MeSH
• proliferace buněk * účinky léků   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• viabilita buněk * účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• diabetes mellitus 2. typu genetika   MeSH
• fibrinogen genetika   MeSH
• genetické markery genetika   MeSH
• infarkt myokardu genetika   MeSH
• integrin alfa2beta1 genetika   MeSH
• lidé MeSH
• polymorfismus genetický genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• srovnávací studie MeSH

Platelets of full-term newborns and those of healthy adult donors were compared for constitutive expression of surface glycoproteins (GP) Ia-IIa, GP Ib, GP IIb-IIIa, and GP IV and for their activation responses to an agonist by detection of surface expression of activation markers P-selectin and CD63. Resting neonatal platelets showed significantly lower expression of GP Ia-IIa, GP Ib, and GP IIb-IIIa. In contrast, the expression of GP IV was significantly higher compared with platelets of adults. The expression of activation markers P-selectin and CD63 was assessed after in vitro activating of platelets with 0-15 microM human thrombin receptor-activating peptide. At low concentrations of thrombin receptor-activating peptide, the extent of surface expression of activation markers did not differ significantly between adult and neonatal platelets. However, after activation with 15 microM thrombin receptor-activating peptide, the extent of surface expression of P-selectin and CD63 was significantly lower in neonatal platelets. Because of ethical reasons, our study was conducted on neonates with a moderate neonatal hyperbilirubinemia. The remote possibility that hyperbilirubinemia could influence the expression of platelet surface receptors and the reactivity of neonatal platelet cannot be excluded. The role of higher expression of GP IV on neonatal platelets, also seen in certain hematologic malignancies in adults, remains to be elucidated. The lower expression of platelet adhesive receptors and the limited ability to up-regulate granular glycoproteins may play a role in the impairment of function of neonatal platelets.

• MeSH
• aktivace trombocytů účinky léků   MeSH
• antigeny CD63 MeSH
• CD antigeny krev   MeSH
• dospělí MeSH
• glykoproteiny membrány trombocytů * metabolismus   MeSH
• integrin alfa2beta1 krev   MeSH
• lidé MeSH
• novorozenec MeSH
• P-selektin krev   MeSH
• peptidové fragmenty * farmakologie   MeSH
• techniky in vitro MeSH
• trombocytový glykoproteinový komplex Ib-IX metabolismus   MeSH
• trombocytový glykoproteinový komplex IIb-IIIa metabolismus   MeSH
• trombocyty * metabolismus   účinky léků   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• novorozenec MeSH
• Publikační typ
• práce podpořená grantem MeSH
• srovnávací studie MeSH