Ethanol Induction of Innate Immune Signals Across BV2 Microglia and SH-SY5Y Neuroblastoma Involves Induction of IL-4 and IL-13

Lawrimore, Colleen J.; Coleman, Leon G.; Zou, Jian; Crews, Fulton T.

doi:10.3390/brainsci9090228

Cited by 10 publications

(6 citation statements)

References 71 publications

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“…BV2 cells were seeded (5 × 10 5 cells per well) in the lower layer of six-well plates (723101, NEST, China). After precipitation for 30 min, APPswe cells (2.5 × 10 5 cells per well) or Aβ 25–35 -supplemented SH-SY5Y cells (2.5 × 10 5 cells per well) were plated on the upper layer of the Transwell (Zujovic and Taupin, 2003 ; Chan et al, 2019 ; Lawrimore et al, 2019 ). Then, the cells were cocultured for 24 h for follow-up experiments.…”

Section: Methodsmentioning

confidence: 99%

Enhanced Autolysosomal Function Ameliorates the Inflammatory Response Mediated by the NLRP3 Inflammasome in Alzheimer’s Disease

Zhou

Deng

Zhao

et al. 2021

Front. Aging Neurosci.

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The pathogenesis of Alzheimer’s disease (AD) involves activation of many NLRP3 inflammatory bodies, which may be related to amyloid β peptide and aggregation of misfolded proteins. Autophagy is an important regulator of inflammatory bodies. However, autophagy shows dynamic changes in the development of AD, and its role in inflammation remains controversial. In this study, the key link between autophagic disorders and the NLRP3 inflammasome in AD was investigated. APP/PS1 double transgenic mice and C57 mice with Aβ25–35 injected into the lateral ventricle were used as two animal models of AD. Immunofluorescence staining and Western blot analysis showed that NLRP3 inflammasome-related proteins and inflammatory cytokines, such as IL-1α, IL-1β, IL-6, IL-12, and TNF-α, were increased and microglia were activated in the brains of both AD animal models. Endogenous overexpression of the APPswe gene and exogenous addition of Aβ25–35 increased the expression of NLRP3 inflammasome-related proteins, while exogenous Aβ25–35 intervention more significantly activated inflammation. Furthermore, LC3 was increased in the AD animal and cell models, and the level of Lamp1 decreased. After overexpression of the primary regulator of lysosomal biogenesis, TFEB, the lysosome protein Lamp1 was increased, and LC3 and inflammatory protein expression were decreased. These results suggest that the NLRP3 inflammasome-mediated inflammatory response is activated in AD animal and cell models, which may be related to the decline in autolysosome function. Overexpression of the TFEB protein can reduce the inflammatory response by improving autolysosome function in AD model cells.

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Section: Methodsmentioning

confidence: 99%

Enhanced Autolysosomal Function Ameliorates the Inflammatory Response Mediated by the NLRP3 Inflammasome in Alzheimer’s Disease

Zhou

Deng

Zhao

et al. 2021

Front. Aging Neurosci.

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“…BV2 microglia (from ICLC #ATL03001; Genoa, Italy) were maintained in culture in standard cell culture conditions as we and others have described previously (Coleman, Zou, Qin, et al, 2017;Lawrimore et al, 2019). Briefly, 3 × 10 5 cells were plated per well in 6-well culture plates in DMEM with 10% FBS, 1× glutaMAX, and 1× penicillin/ streptomycin antibiotic (Life Technologies).…”

Section: Bv2 Microglial Culturesmentioning

confidence: 99%

Extracellular microvesicles promote microglia‐mediated pro‐inflammatory responses to ethanol

Crews

Zou

Coleman

2021

J of Neuroscience Research

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Alcohol use disorder (AUD) pathology features pro-inflammatory gene induction and microglial activation. The underlying cellular processes that promote this activation remain unclear. Previously considered cellular debris, extracellular vesicles (EVs) have emerged as mediators of inflammatory signaling in several disease states.We investigated the role of microvesicles (MVs, 50 nm-100 µm diameter EVs) in pro-inflammatory and microglial functional gene expression using primary organotypic brain slice culture (OBSC). Ethanol caused a unique immune gene signature that featured: temporal induction of pro-inflammatory TNF-α and IL-1β, reduction of homeostatic microglia state gene Tmem119, progressive increases in purinergic receptor P2RY12 and the microglial inhibitory fractalkine receptor CX3CR1, an increase in the microglial presynaptic gene C1q, and a reduction in the phagocytic gene TREM2. MV signaling was implicated in this response as reduction of MV secretion by imipramine blocked pro-inflammatory TNF-α and IL-1β induction by ethanol, and ethanol-conditioned MVs (EtOH-MVs) reproduced the ethanol-associated immune gene signature in naïve OBSC slices. Depletion of microglia prior to ethanol treatment prevented pro-inflammatory activity of EtOH-MVs, as did incubation of EtOH-MVs with the HMGB1 inhibitor glycyrrhizin. Ethanol caused HMGB1 secretion from cultured BV2 microglia in MVs through activation of PI3 kinase. In summary, these studies find MVs modulate pro-inflammatory gene induction and microglial activation changes associated with ethanol. Thus, MVs may represent a novel therapeutic target to reduce neuroinflammation in the setting of alcohol abuse or other diseases that feature a neuroimmune component. [Correction added on 5 April 2021, after first online publication: The copyright line was changed.] K E Y W O R D S alcohol use disorder, extracellular vesicles, inflammation, microglia, neuroimmuneThis is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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“…As introduced above, exposure to high doses of alcohol can induce ROS generation and oxidative stress, but it is unknown whether alcohol-induced oxidative stress can induce cell death in microglial cells. Therefore, we investigated the effects of exposure to EtOH for 24 h at concentrations (10–300 mM) which have been commonly used for in vitro studies [ 34 , 35 , 37 , 38 , 39 ]. As shown in Figure 3 a,b, exposure to EtOH induced concentration-dependent microglial cell death, with the cell death level significantly increased following exposure to high concentrations (100 and 300 mM).…”

Section: Resultsmentioning

confidence: 99%

“…Recent studies from our group and other authors using pharmacological and/or genetic interventions have shown an important role for the TRPM2 channel in signaling mechanisms mediating the responses of microglial cells, including cell death, to ROS and pathological factors, such as amyloid β-peptide, MPP + , and Zn 2+ , which are known to be able to stimulate ROS generation [ 25 , 26 , 31 ]. In this study, we are interested in whether alcohol-induced oxidative stress results in microglial cell death and, if this is true, whether and how the TRPM2 channel is involved in such alcohol-induced cell death, using murine BV2 microglial cells, a widely-used cell model for in vitro studies of microglial cell-mediated mechanisms, including those induced by ethanol [ 34 , 35 ]. We show that exposure to high doses of EtOH induces microglial cell death and provide evidence to show that EtOH causes microglial cell death via induction of NOX-mediated ROS generation and subsequent activation of PARP and the TRPM2 channel.…”

Section: Introductionmentioning

confidence: 99%

Ethanol Induces Microglial Cell Death via the NOX/ROS/PARP/TRPM2 Signalling Pathway

et al. 2020

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Microglial cells are the primary immune cell resident in the brain. Growing evidence indicates that microglial cells play a prominent role in alcohol-induced brain pathologies. However, alcohol-induced effects on microglial cells and the underlying mechanisms are not fully understood, and evidence exists to support generation of oxidative stress due to NADPH oxidases (NOX_-mediated production of reactive oxygen species (ROS). Here, we investigated the role of the oxidative stress-sensitive Ca2+-permeable transient receptor potential melastatin-related 2 (TRPM2) channel in ethanol (EtOH)-induced microglial cell death using BV2 microglial cells. Like H2O2, exposure to EtOH induced concentration-dependent cell death, assessed using a propidium iodide assay. H2O2/EtOH-induced cell death was inhibited by treatment with TRPM2 channel inhibitors and also treatment with poly(ADP-ribose) polymerase (PARP) inhibitors, demonstrating the critical role of PARP and the TRPM2 channel in EtOH-induced cell death. Exposure to EtOH, as expected, led to an increase in ROS production, shown using imaging of 2’,7’-dichlorofluorescein fluorescence. Consistently, EtOH-induced microglial cell death was suppressed by inhibition of NADPH oxidase (NOX) as well as inhibition of protein kinase C. Taken together, our results suggest that exposure to high doses of ethanol can induce microglial cell death via the NOX/ROS/PARP/TRPM2 signaling pathway, providing novel and potentially important insights into alcohol-induced brain pathologies.

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Ethanol Induction of Innate Immune Signals Across BV2 Microglia and SH-SY5Y Neuroblastoma Involves Induction of IL-4 and IL-13

Cited by 10 publications

References 71 publications

Enhanced Autolysosomal Function Ameliorates the Inflammatory Response Mediated by the NLRP3 Inflammasome in Alzheimer’s Disease

Enhanced Autolysosomal Function Ameliorates the Inflammatory Response Mediated by the NLRP3 Inflammasome in Alzheimer’s Disease

Extracellular microvesicles promote microglia‐mediated pro‐inflammatory responses to ethanol

Ethanol Induces Microglial Cell Death via the NOX/ROS/PARP/TRPM2 Signalling Pathway

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