Taurine Chloramine decreases cell viability and cytokine production in blood and spleen lymphocytes from septic rats

AbstractEmerging evidence suggests that microglia can support neurogenesis. Little is known about the mechanisms by which microglia regulate the cortical environment and stimulate cortical neurogenesis. We used an in vitro co-culture model system to investigate the hypothesis that microglia respond to soluble signals from cortical cells, particularly following mechanical injury, to alter the cortical environment and promote cortical cell proliferation, differentiation, and survival. Analyses of cortical cell proliferation, cell death, neurogenic protein expression, and intracellular signaling were performed on uninjured and injured cortical cells in co-culture with microglial cell lines. Microglia soluble cues enhanced cortical cell viability and proliferation cortical cells. Co-culture of injured cortical cells with microglia significantly reduced cell death of cortical cells. Microglial co-culture significantly increased Nestin + and α-internexin + cortical cells. Multiplex ELISA and RT-PCR showed decreased pro-inflammatory cytokine production by microglia co-cultured with injured cortical cells. Inhibition of AKT phosphorylation in cortical cells blocked microglial-enhanced cortical cell viability and expression of neurogenic markers in vitro. This in vitro model system allows for assessment of the effect of microglial-derived soluble signals on cortical cell viability, proliferation, and stages of differentiation during homeostasis or following mechanical injury. These data suggest that microglia cells can downregulate inflammatory cytokine production following activation by mechanical injury to enhance proliferation of new cells capable of neurogenesis via activation of AKT intracellular signaling. Increasing our understanding of the mechanisms that drive microglial-enhanced cortical neurogenesis during homeostasis and following injury in vitro will provide useful information for future primary cell and in vivo studies.

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Evaluation of cell damage and modulation of cytokines TNF-α, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy

Brazilian Journal of Biology ◽

10.1590/1519-6984.193748 ◽

2020 ◽

Vol 80 (3) ◽

pp. 497-505

Author(s):

R. D. R. Tiveron ◽

D. A. Costa ◽

M. D. I. Leite ◽

C. B. S. Vaz ◽

M. Sousa ◽

...

Keyword(s):

Photodynamic Therapy ◽

Immune Responses ◽

Cell Viability ◽

Cytokine Production ◽

Cell Damage ◽

Protoporphyrin Ix ◽

Fold Increase ◽

Dna Degradation ◽

Tnf Α

Abstract Little is known regarding whether photodynamic therapy (PDT)-induced cell death can substantially compromise macrophages (MΦ), which are important cells in PDT-induced immune responses. Here, parameters of PDT-mediated MΦ cytotoxicity and cytokine production in response to protoporphyrin IX (PpIX) were evaluated. Peritoneal MΦ from BALB/c mice were stimulated in vitro with PDT, light, PpIX, or lipopolysaccharide (LPS). After that, cell viability, lipid peroxidation, Nitric Oxide (NO), DNA damage, TNF-α, IL-6 and IL-10 were evaluated. Short PDT exposure reduced cell viability by 10–30%. There was a two-fold increase in NO and DNA degradation, despite the non-increase in lipoperoxidation. PDT increased TNF-α and IL-10, particularly in the presence of LPS, and decreased the production of IL-6 to 10-fold. PDT causes cellular stress, induces NO radicals and leads to DNA degradation, generating a cytotoxic microenvironment. Furthermore, PDT modulates pro- and anti-inflammatory cytokines in MΦ.

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Impact of cell culture methods on the outcomes of the in vitro inflammatory response in nasal polyps

Rhinology Journal ◽

10.4193/rhino11.036 ◽

2011 ◽

Vol 49 (5) ◽

pp. 562-569

Author(s):

L. Fernández-Bertolín ◽

J. Mullol ◽

I. Alobid ◽

J. Roca-Ferrer ◽

C. Picado ◽

...

Keyword(s):

Cell Viability ◽

Nasal Mucosa ◽

Nasal Polyp ◽

Cytokine Production ◽

Culture Conditions ◽

Culture Methods ◽

Gm Csf ◽

Research Outcomes ◽

Cytokine Levels

Background: In vitro culture of nasal polyp cells is frequently used in the investigation of inflammatory mechanisms and effect of treatments in nasal polyposis. Research outcomes may, however, be influenced by the culture methodology used. Methods: Nasal polyp and nasal mucosa in vitro fibroblast cultures were pre-treated with foetal bovine serum (FBS)-free culture medium or medium supplemented with either FBS or charcoal-stripped (cs) FBS. Cells were then stimulated with FBS or csFBS, with or without different doses of dexamethasone for 4 and 24h. IL-6, IL-8, GM-CSF and VEGF release and cell viability were measured. Results: The highest cytokine levels were found in growth-arrested cells stimulated with 10% FBS. csFBS poorly stimulated cytokine release. Nasal polyp released larger IL-8 amounts than nasal mucosa fibroblasts. Dexamethasone decreased cytokine production dose- and time-dependently in both nasal mucosa and nasal polyp fibroblasts. The IC25 of IL-8 inhibition by dexamethasone was higher in nasal polyp than in nasal mucosa fibroblasts. Cell viability did not differ among treatments. Conclusions: Cytokine production by in vitro cultured nasal fibroblasts is affected by the culture conditions used and is inhibited by dexamethasone in both fibroblast types. Our results highlight the importance of culture methodology on nasal polyp research outcomes.

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Ginsenoside Rb1 alleviates lipopolysaccharide-induced inflammatory injury by downregulating miR-222 in WI-38 cells

10.21203/rs.2.24163/v1 ◽

2020 ◽

Author(s):

Erhu Wei ◽

Xiao Fang ◽

Peisheng Jia ◽

Mingxia Li ◽

Peina Jin ◽

...

Keyword(s):

Cell Viability ◽

Inflammatory Cytokine ◽

Molecular Mechanisms ◽

Cytokine Production ◽

Panax Notoginseng ◽

Lung Fibroblasts ◽

Ginsenoside Rb1 ◽

Inflammatory Cytokine Production ◽

Inflammatory Injury ◽

Factor Α

Abstract Background Pneumonia is a serious respiratory tract infection disease in children, which threatens to the health or life of children patients. Ginsenoside Rb1 (Rb1) is a principle active ingredient extracted from the root of Panax notoginseng (Burk.) F.H.Chen with anti-inflammatory effect. Our study aimed to determine the effects and molecular mechanisms of Rb1 on lipopolysaccharide (LPS)-induced inflammatory injury of lung fibroblasts WI-38 cells. Methods Cell viability and apoptosis were evaluated by CCK-8 and flow cytometry analysis, respectively. The production of inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin (IL)-lβ, and IL-6 were measured by ELISA and RT-qPCR. miR-222 expression was examined by RT-qPCR. The expression levels of the nuclear factor-kappa B (NF-κB) p65 and phosphorylated p65 were detected by western blot. Results LPS stimulation induced WI-38 cell inflammatory injury by inhibiting cell viability, and inducing apoptosis and inflammatory cytokine production, while treatment with Rb1 significantly attenuated LPS-induced inflammatory injury in WI-38 cells. Additionally, Rb1 decreased LPS-induced upregulation of miR-222 and activation of the NF-κB pathway in WI-38 cells. Overexpression of miR-222 abolished the inhibitory effects of Rb1 on LPS-induced viability reduction, apoptosis, inflammatory cytokine production and activation of the NF-κB pathway. Conclusion Rb1 alleviated LPS-induced inflammatory injury in WI-38 cells via downregulating miR-222 and inactivation of the NF-κB pathway, contributing to our understanding of the effects and molecular mechanism of Rb1 in pneumonia.

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