Journal: Journal of Neuroinflammation

Article Title: A selective inhibitor of the NLRP3 inflammasome as a potential therapeutic approach for neuroprotection in a transgenic mouse model of Huntington’s disease

doi: 10.1186/s12974-022-02419-9

Figure Lengend Snippet: MCC950 markedly reduces cytotoxicity in striatal progenitor cells and BV2 microglial cells. A , B BV2 microglia were incubated for 4 h with LPS (1 μg/mL) followed by incubation with MCC950 (1 μM) for 2 h. The cells were then incubated with ATP (1 mM, 24 h). Total lysates of BV2 microglial cells were assessed by Western blot analysis to determine the levels of the NLRP3 and actin proteins. The molecular mass is indicated in kilodaltons. C , D BV2 microglia were incubated for 4 h with LPS (1 μg/mL) followed by incubation with MCC950 (1 μM) for 2 h. The cells were then incubated with ATP (1 mM for 24 h). Cell survival ( C ) and IL-1β expression levels ( D ) were measured using the CCK-8 assay and ELISA, respectively. The values of the indicated cells were normalized to those of untreated BV2 cells. * P < 0.05 compared to LPS/ATP treated cells ( n = 3). E , F ST Hdh Q109 cells were incubated for 24 h with MCC950 (1 μM). Total lysates of ST Hdh Q7 and ST Hdh Q109 cells were assessed using Western blot analysis. G ST Hdh Q7 and ST Hdh Q109 cells were incubated for 24 h with MCC950 (1 μM). Cell death was quantified using the CCK-8 assay; the values of the indicated cells were normalized to those of untreated ST Hdh Q7 cells. The data are presented as the mean ± SEM from three independent experiments. * P < 0.05, ST Hdh Q7 vs. ST Hdh Q109 cells; # P < 0.05 vs. untreated ST Hdh Q109 cells. H BV2 cells were incubated with LPS (1 µg/mL for 4 h) with or without MCC950 for 2 h before stimulation with ATP (1 mM for 24 h). The BV2 medium was then collected and used to culture the ST Hdh Q109 cells for an additional 24 h. ST Hdh Q7 and ST Hdh Q109 cell viability was determined by CCK-8 assay. The data are presented as the mean ± SEM from three independent experiments. * P < 0.05, ST Hdh Q7 vs. ST Hdh Q109 cells; # P < 0.05 vs. untreated ST Hdh Q109 cells. I BV2 cells were treated with or without MCC950 (1 μM) and 3-NP (5, 10, and 20 mM) for 24 h. The cell viability was determined using the CCK-8 assay. Data are presented as the mean ± SEM from three independent experiments. * P < 0.05 compared with controls ( n = 3)

Article Snippet: BV2 cells (mouse, C57BL/6; brain, microglial cells) were purchased from the American Type Culture Collection (Rockville, MD).

Techniques: Incubation, Western Blot, Expressing, CCK-8 Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Fungi

Article Title: T-2 Toxin Induces Apoptotic Cell Death and Protective Autophagy in Mouse Microglia BV2 Cells

doi: 10.3390/jof8080761

Figure Lengend Snippet: Cytotoxic effects of T-2 toxin on BV2 cells. BV2 cells were treated with T-2 toxin at the final doses of 0.625, 1.25, 2.5, 5, and 10 ng/mL for 6, 12, and 24 h, respectively; cell viabilities were measured. All data are shown as the mean ± SD from three independent experiments ( n = 3). ** p < 0.01, compared to the control group.

Article Snippet: Mouse BV2 microglia were purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Control

Journal: Journal of Fungi

Article Title: T-2 Toxin Induces Apoptotic Cell Death and Protective Autophagy in Mouse Microglia BV2 Cells

doi: 10.3390/jof8080761

Figure Lengend Snippet: T-2 toxin induces apoptotic cell death in BV2 cells. BV2 cells were treated with T-2 toxin at final concentrations of 1.25, 2.5, and 5 ng/mL for 24 h. Apoptotic rates of T-2 toxin-treated BV2 cells were analyzed by flow cytometry following annexin V-FITV/PI staining. Q1, necrosis cells; Q2, later apoptotic cells; Q3, early apoptotic cells; Q4, live cells. The results are presented as the mean ± SD from three independent experiments ( n = 3). * p < 0.05 and ** p < 0.01, compared to the control group.

Article Snippet: Mouse BV2 microglia were purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Flow Cytometry, Staining, Control

Journal: Journal of Fungi

Article Title: T-2 Toxin Induces Apoptotic Cell Death and Protective Autophagy in Mouse Microglia BV2 Cells

doi: 10.3390/jof8080761

Figure Lengend Snippet: T-2 toxin treatment induces the production of ROS in BV2 cells. ( A ) Cells were treated with T-2 toxin at the final concentrations of 0, 1.25, 2.5, and 5 ng/mL for 24 h; the levels of intracellular ROS were measured by staining with DCFH-DA. The representative images (on the left) were selected (on the left) and quantitative analysis (on the right) was performed. Bar = 20 μm. ( B ) The levels of MDA. ( C ) The levels of CAT activities. ( D ) The levels of SOD activities. ( E ) Cells were pre-treated with NAC at 2.5 mM for 2 h, then co-treated with or without T-2 toxin at 2.5 ng/mL for 24 h; intracellular ROS levels were detected using flow cytometer. ( F ) The effects of NAC pretreatment on T-2 toxin-induced loss of cell viability. Cell treatment is same as in ( E ). All results are presented as mean ± SD from three independent experiments ( n = 3). * p < 0.05 and ** p < 0.01, compared to the control.

Article Snippet: Mouse BV2 microglia were purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Staining, Flow Cytometry, Control

Journal: Journal of Fungi

Article Title: T-2 Toxin Induces Apoptotic Cell Death and Protective Autophagy in Mouse Microglia BV2 Cells

doi: 10.3390/jof8080761

Figure Lengend Snippet: T-2 toxin induces loss of mitochondrial membrane potential in BV2 cells. Cells were treated with T-2 toxin at the final concentrations of 1.25, 2.5, and 5 ng/mL for 24 h, then stained with Rh123 and observed by a fluorescence microscope. The representative images (on the left) were selected and quantitative analysis (on the right) was performed. Bar = 20 μm. Results are presented as mean ± SD from three independent experiments ( n = 3). ** p < 0.01, compared to the control.

Article Snippet: Mouse BV2 microglia were purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Membrane, Staining, Fluorescence, Microscopy, Control

Journal: Journal of Fungi

Article Title: T-2 Toxin Induces Apoptotic Cell Death and Protective Autophagy in Mouse Microglia BV2 Cells

doi: 10.3390/jof8080761

Figure Lengend Snippet: T-2 toxin treatment activates cell autophagy and autophagy play a protective role. ( A ) BV2 cells were treated with T-2 toxin at 1.25, 2.5, and 5 ng/mL for 24 h; the expressions of Beclin1 and LC3II proteins were measured by Western blot. The representative bands (on the left) and quantitative analysis (on the right) are shown. ( B ) BV2 cells were pre-treated with chloroquine (CQ) at 5 μM for 1 h, then co-treated with T-2 toxin at 2.5 ng/mL for an additional 24 h; the expression of LC3II protein was measured. ( C ) Autophagy flux was monitored by the mRFP-GFP-LC3 plasmid transfection method. Cells were treated with Q at 5 μM, T-2 toxin at 2.5 ng/mL, or co-treatment for 12 h; autophagosomes and autolysosomes were observed and photographed by a laser scanning confocal microscope. Bar = 10 μm. ( D – F ) Same treatments in BV2 cells as in B; cell viabilities ( D ), caspase-3 activities ( E ), and cell apoptotic rates ( F ) were measured. The results are shown as the means ± SD from three independent experiments ( n = 3). * p < 0.05 and ** p < 0.01, compared to the control; # p < 0.05, and ## p < 0.01, compared to the T-2 alone treatment group.

Article Snippet: Mouse BV2 microglia were purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Western Blot, Expressing, Plasmid Preparation, Transfection, Microscopy, Control

Journal: Journal of Fungi

Article Title: T-2 Toxin Induces Apoptotic Cell Death and Protective Autophagy in Mouse Microglia BV2 Cells

doi: 10.3390/jof8080761

Figure Lengend Snippet: Schematic diagram of the proposed mechanisms of T-2 toxin-induced apoptosis in mouse microglia BV2 cells. ROS, reactive oxygen species; MMP, mitochondrial membrane potential; PARP-1, poly (ADP-ribose) polymerase-1; MDA, malondialdehyde; CAT, catalase; SOD, dismutase; HO-1, heme oxygenase-1.

Article Snippet: Mouse BV2 microglia were purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Membrane

Journal: Journal of Neuroinflammation

Article Title: TREM-2 - p38 MAPK signaling regulates neuroinflammation during chronic cerebral hypoperfusion combined with diabetes mellitus

doi: 10.1186/s12974-019-1688-9

Figure Lengend Snippet: Characterization of TREM2 expression in different groups of BV2 cells. BV2 cells were transfected with TREM2 lentiviral particles or TREM2-specific siRNAs. The relative levels of TREM2 expression in different groups were determined by quantitative RT-PCR and Western blot. a Quantitative RT-PCR analysis of TREM2-specific siRNA mRNA transcripts. b Western blot analysis of TREM2 expression in BV2 cells transfected with TREM2-specific siRNAs. c Quantitative RT-PCR analysis of TREM2 mRNA transcripts. d Western blot analysis of TREM2 expression in BV2 cells transfected with TREM2 lentiviral particles. ** P < 0.01; *** P < 0.001 vs. the BV2 cells

Article Snippet: Immortalized mouse BV2 microglial cell lines were purchased from ScienCell (CA, USA) and cultured in MEM medium with 10% FBS and 1% penicillin/streptomycin.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot

Journal: Journal of Neuroinflammation

Article Title: TREM-2 - p38 MAPK signaling regulates neuroinflammation during chronic cerebral hypoperfusion combined with diabetes mellitus

doi: 10.1186/s12974-019-1688-9

Figure Lengend Snippet: TREM-2 knockdown enhanced while TREM-2 overexpression inhibited pro-inflammatory cytokines in high glucose-hypoxia-stimulated microglia by targeting p38 MAPK. a Representative bands of Western blot data. b Quantitative analysis of the Western blot bands. c , d The mRNA expression of pro-inflammatory cytokines (IL-1β, TNF-α, iNOS, and COX2) was detected by RT-PCR. ** P < 0.01; *** P < 0.001 vs. the BV2 cells (Normal); # P < 0.05; ## P < 0.01; ### P < 0.001 vs. the BV2 cells (hypoxia and high glucose)

Article Snippet: Immortalized mouse BV2 microglial cell lines were purchased from ScienCell (CA, USA) and cultured in MEM medium with 10% FBS and 1% penicillin/streptomycin.

Techniques: Knockdown, Over Expression, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Aging Cell

Article Title: Human Umbilical Cord Mesenchymal Stem Cells Ameliorate Cognitive Decline by Restoring Senescent Microglial Function via NF ‐ κB ‐ SREBP1 Pathway Inhibition

doi: 10.1111/acel.70259

Figure Lengend Snippet: hUC‐MSCs attenuate H 2 O 2 ‐induced microglial senescence, reduce LD accumulation and restore phagocytic function. (a) Schematics of BV2 cells treatment. (b) Representative image of SA‐β‐gal staining in BV2 cells and statistical analysis of the proportion of positive cells, n = 3 per group. (c) The protein levels of p21 and γ‐H2AX were determined by western blot and quantified using ImageJ software. n = 3 per group. (d‐e) p16 and SASP mRNA levels were determined by qPCR and quantified using ImageJ software. n = 3 per group. (f) Representative image of BODIPY (green) in BV2 cells and (i) the mean fluorescence intensity of BODIPY was quantified by ImageJ software. n = 3 per group. (g) Representative image of PLIN2 (green) in BV2 cells and (j) the mean fluorescence intensity of PLIN2 was quantified by ImageJ software. n = 3 per group. (h) Representative image of zymosan (green) in BV2 cells and (k) the mean fluorescence intensity of zymosan was quantified by ImageJ software. n = 3 per group. Data are presented as means ± SD. Statistical significance was determined by One‐way ANOVA, following normality and lognormality tests. ns, no significant, * p < 0.05, ** p < 0.01.

Article Snippet: BV2 cells (a mouse microglia cell line) and HT22 cells (a mouse neuron cell line) were purchased from Procell Life Science & Technology Co. Ltd. and cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco) supplemented with 10% Fetal Bovine Serum (FBS, Invitrogen) in an atmosphere of 5% CO 2 at 37°C. hUC‐MSCs were supplied by the National Engineering Research Center of Human Stem Cells and the Guangxiu Tech Biotechnology Co.

Techniques: Staining, Western Blot, Software, Fluorescence

Journal: Aging Cell

Article Title: Human Umbilical Cord Mesenchymal Stem Cells Ameliorate Cognitive Decline by Restoring Senescent Microglial Function via NF ‐ κB ‐ SREBP1 Pathway Inhibition

doi: 10.1111/acel.70259

Figure Lengend Snippet: hUC‐MSCs alleviate senescent microglia‐induced damage to aging neurons. (a) Schematics of HT22 cells treatment. (b) qPCR analysis of p21 mRNA levels ( n = 5 per group). (c, d) Representative SA‐β‐gal staining image of HT22 cells and quantification of SA‐β‐gal positive HT22 cells ( n = 3 per group). (e) qPCR analysis of tnf‐α ( n = 4 per group) and il‐6 ( n = 3 per group) mRNA levels. (f) Representative flow cytometry plots and quantification of HT22 cell apoptosis ( n = 3 per group). Data are presented as means ± SD. Statistical significance was determined by one‐way ANOVA, following normality and lognormality tests. ns = no significant, * p < 0.05, ** p < 0.01. B‐H group: Senescent microglia–neuron co‐culture group (BV2‐H 2 O 2 /HT22‐H 2 O 2 ): BV2 cells were treated with H 2 O 2 for 24 h, then co‐cultured with H 2 O 2 ‐induced senescent HT22 neurons for another 24 h. B‐M group: hUC‐MSC‐preconditioned senescent microglia–neuron co‐culture group (BV2‐H 2 O 2 ‐MSC/HT22‐H 2 O 2 ): Senescent BV2 cells (H 2 O 2 ‐treated) were co‐cultured with hUC‐MSCs for 24 h, then co‐cultured with senescent HT22 neurons for an additional 24 h.

Article Snippet: BV2 cells (a mouse microglia cell line) and HT22 cells (a mouse neuron cell line) were purchased from Procell Life Science & Technology Co. Ltd. and cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco) supplemented with 10% Fetal Bovine Serum (FBS, Invitrogen) in an atmosphere of 5% CO 2 at 37°C. hUC‐MSCs were supplied by the National Engineering Research Center of Human Stem Cells and the Guangxiu Tech Biotechnology Co.

Techniques: Staining, Flow Cytometry, Co-Culture Assay, Cell Culture

Journal: Aging Cell

Article Title: Human Umbilical Cord Mesenchymal Stem Cells Ameliorate Cognitive Decline by Restoring Senescent Microglial Function via NF ‐ κB ‐ SREBP1 Pathway Inhibition

doi: 10.1111/acel.70259

Figure Lengend Snippet: JSH‐23 reduces LD accumulation, enhances phagocytic function, and attenuates H 2 O 2 ‐induced senescence in microglia. (a) Schematics of BV2 cells treatment. (b) Representative image of p65 (green) in BV2 cells and (c) statistical analysis of p65 nuclear‐positive cells. n = 3 per group. (d) The protein levels of p65, p‐p65 and SREBP1 were determined by western blot and quantified using ImageJ software. n = 3 per group. (e) Representative image of SA‐β‐gal in BV2 cells and (f) statistical analysis of positive cells. (g) The SASP mRNA levels were determined and quantified by qPCR. n = 3 per group. (h) Representative flow cytometry plots of BODIPY in BV2 cells. n = 3 per group. (i) Representative image of PLIN2 (green) in BV2 cells and mean fluorescence intensity quantified using ImageJ software. n = 3 per group. (j) Representative image of zymosan (green) in BV2 cells and mean fluorescence intensity quantified using ImageJ software. n = 3 per group. Data are presented as means±SD. Statistical significance was determined by One‐way ANOVA, following normality and lognormality tests. ns, no significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BV2 cells (a mouse microglia cell line) and HT22 cells (a mouse neuron cell line) were purchased from Procell Life Science & Technology Co. Ltd. and cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco) supplemented with 10% Fetal Bovine Serum (FBS, Invitrogen) in an atmosphere of 5% CO 2 at 37°C. hUC‐MSCs were supplied by the National Engineering Research Center of Human Stem Cells and the Guangxiu Tech Biotechnology Co.

Techniques: Western Blot, Software, Flow Cytometry, Fluorescence

Journal: Neuropathology and Applied Neurobiology

Article Title: Microglia induce an interferon‐stimulated gene expression profile in glioblastoma and increase glioblastoma resistance to temozolomide

doi: 10.1111/nan.13016

Figure Lengend Snippet: Mouse BV2 immortalised microglia reduce TMZ sensitivity of glioblastoma spheroids. ( A–C ) Illustration of culture set‐ups showing direct co‐culture of microglia and glioblastoma (GBM) spheroids (A), indirect co‐culture with GBM spheroids and microglia separated by a microporous membrane (B) and a set‐up where GBM spheroids are treated with conditioned medium derived from microglia cultures (C). ( D ) Images of T87 spheroids cultured without (top) and with (bottom) BV2 microglia in a direct co‐culture set‐up with 0, 60 and 120 μM TMZ. The left panels show light microscope images of the spheroids, and the right panels indicate fluorescent microscope images of their respective propidium iodide (PI) signals (outlined using ImageJ). ( E, F ) Two spheroid cultures (T87 and T78) treated with TMZ showed reduced cell death (reduced PI intensity) after direct co‐culture with BV2 microglia. ( G, H, I ) Three spheroid cultures (T87, T78 and T121) treated with TMZ showed reduced cell death after indirect co‐culture with BV2 microglia. ( J, K, L ) Three spheroid cultures (T87, T78 and T121) treated with TMZ and control medium (COM, with and without TMZ) or BV2 microglia‐conditioned medium (MCM, with and without TMZ). After 24 h (T87) or 48 h (T78 and T121) of culture, a significantly reduced TMZ‐induced cell death of GBM spheroids was observed in MCM cultures treated with 0 or 60 μM TMZ, but not 120 μM TMZ. In figures J–L the TMZ concentrations right below the x‐axis indicate TMZ in the culture, whereas the TMZ concentrations below the brackets indicate TMZ in the pre‐treated COM and MCM cultures. The level of significance is indicated with asterisks (* p < 0.05; ** p < 0.01; *** p < 0.005). The standard error of the mean (SEM) based on at least two biological replicates is indicated by the vertical lines. The scale bar in D is 100 μm.

Article Snippet: The mouse BV2 immortalised microglia cell line [ , ] (AcceGen Biotech, NJ, USA) was maintained in RPMI 1640 Medium with GlutaMAXTM (Gibco) supplemented with 5% foetal bovine serum (FBS) (Sigma‐Aldrich) and 1% penicillin/streptomycin.

Techniques: Co-Culture Assay, Membrane, Derivative Assay, Cell Culture, Light Microscopy, Microscopy, Control

Journal: Food Science & Nutrition

Article Title: A mixture of extracts from natural ingredients reduces the neurotoxic polarization of microglia via modulating NF‐κB / NF‐E2 ‐related factor 2 activation

doi: 10.1002/fsn3.4045

Figure Lengend Snippet: The mixture regulated the gene expression and NF‐κB/Nrf2 activation in BV2 microglia during exposure to Aβ and PgLPS. The mRNA expression of TNF‐α (a), IL‐1β (b) at 1 h after exposure to Aβ and PgLPS (AL) with or without pretreatment with propolis (P) or mixture (Mix). (c) Immunofluorescent CLMS images indicating the nuclear translocation of p65 (green) in BV2 cells with Hoechst‐stained nuclei (blue) after exposure to AL for 1 h. Scale bar, 10 μm. The mRNA expression of IL‐10 (d) and BDNF (e) at 1 h after exposure to AL with or without pretreatment with propolis (P) or mixture (Mix). (f) Immunofluorescent CLMS images indicating the nuclear translocation of Nrf2 (green) in BV2 cells with Hoechst‐stained nuclei (blue) after exposure to AL for 1 h. Scale bar, 10 μm. Each column and bar represents the mean ± SD ( n = 3, each). Asterisks indicate a statistically significant difference from the value in the control group (* p < .05, *** p < .001, one‐way ANOVA). Swords indicate a statistically significant difference from the value in the Aβ and PgLPS‐exposed group ( &&& p < .001, one‐way ANOVA). Hash marks indicate a statistically significant difference from the value in the propolis and mixture group ( ### p < .001, one‐way ANOVA).

Article Snippet: The c‐myc‐immortalized mouse microglial cell line MG6 (Cat: RCB2403) was purchased from RIKEN Cell Bank; BV2 cells (the immortalized mouse microglial cell line) were obtained from ACCEGEN (Cat: ABC‐TC212S).

Techniques: Gene Expression, Activation Assay, Expressing, Translocation Assay, Staining, Control

Journal: Antioxidants

Article Title: Enantioselective Synthesis and Pharmacological Evaluation of Aza-CGP37157–Lipoic Acid Hybrids for the Treatment of Alzheimer’s Disease

doi: 10.3390/antiox11010112

Figure Lengend Snippet: Anti-inflammatory activity of compounds 7a–d against LPS-induced activation of BV2 microglial cells.

Article Snippet: BV2 mouse microglial cells (ATCC; CRL-2469) were cultured in RPMI (SigmaAldrich, R6504), with 100 μg/mL streptomycin, 100 units/mL penicillin and 10% FBS.

Techniques: Activity Assay, Activation Assay