Regulation of Microglia Activity by Glaucocalyxin-A: Attenuation of Lipopolysaccharide-Stimulated Neuroinflammation through NF-κB and p38 MAPK Signaling Pathways
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{"title"=>"Regulation of Microglia Activity by Glaucocalyxin-A: Attenuation of Lipopolysaccharide-Stimulated Neuroinflammation through NF-κB and p38 MAPK Signaling Pathways", "type"=>"journal", "authors"=>[{"first_name"=>"Byung Wook", "last_name"=>"Kim", "scopus_author_id"=>"55584522200"}, {"first_name"=>"Sushruta", "last_name"=>"Koppula", "scopus_author_id"=>"8839545500"}, {"first_name"=>"Seong Su", "last_name"=>"Hong", "scopus_author_id"=>"35074089600"}, {"first_name"=>"Sae Bom", "last_name"=>"Jeon", "scopus_author_id"=>"7203005758"}, {"first_name"=>"Ji Hye", "last_name"=>"Kwon", "scopus_author_id"=>"55584278100"}, {"first_name"=>"Bang Yeon", "last_name"=>"Hwang", "scopus_author_id"=>"56421347600"}, {"first_name"=>"Eun Jung", "last_name"=>"Park", "scopus_author_id"=>"17233933700"}, {"first_name"=>"Dong Kug", "last_name"=>"Choi", "scopus_author_id"=>"7401643837"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84873519871", "doi"=>"10.1371/journal.pone.0055792", "pui"=>"368294422", "pmid"=>"23393601", "scopus"=>"2-s2.0-84873519871", "issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\r1932-6203 (Linking)"}, "id"=>"5ebe4d55-3760-345e-a733-0c084fa40798", "abstract"=>"Microglial cells are the resident macrophages and intrinsic arm of the central nervous system innate immune defense. Microglial cells become activated in response to injury, infection, environmental toxins, and other stimuli that threaten neuronal survival. Therefore, regulating microglial activation may have therapeutic benefits that lead to alleviating the progression of inflammatory-mediated neurodegeneration. In the present study, we investigated the effect of glaucocalyxin A (GLA) isolated from Rabdosia japonica on the production of pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated primary microglia and BV-2 cells. GLA significantly inhibited LPS-induced production of nitric oxide and reversed the morphological changes in primary microglia. Further, GLA suppressed expression of inducible nitric oxide synthase and cyclooxygenase-2 dose-dependently at the mRNA and protein levels. The production of proinflammatory cytokines such as tumor necrosis factor-α, interleukin-1β (IL)-1β, and IL-6 were inhibited by suppressing their transcriptional activity. Furthermore, GLA suppressed nuclear factor-κB activation by blocking degradation of IκB-α and inhibited the induction of lipocalin-2 expression in LPS-stimulated BV-2 cells. Mechanistic study revealed that the inhibitory effects of GLA were accompanied by blocking the p38 mitogen activated protein kinase signaling pathway in activated microglia. In conclusion, given that microglial activation contributes to the pathogenesis of neurodegenerative diseases, GLA could be developed as a potential therapeutic agent for treating microglia-mediated neuroinflammatory diseases.", "link"=>"http://www.mendeley.com/research/regulation-microglia-activity-glaucocalyxina-attenuation-lipopolysaccharidestimulated-neuroinflammat", "reader_count"=>18, "reader_count_by_academic_status"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Ph. D. Student"=>2, "Other"=>3, "Student > Master"=>3, "Student > Bachelor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Ph. D. Student"=>2, "Other"=>3, "Student > Master"=>3, "Student > Bachelor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>7, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>3, "Medicine and Dentistry"=>2, "Neuroscience"=>2, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Neuroscience"=>{"Neuroscience"=>2}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>3}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>7}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/496353"], "description"=>"<p>A: Cells were pre-treated with the indicated doses of GLA for 1 h before LPS (100 ng/mL) treatment. The levels of TNF-α, IL-1β, and IL-6 mRNA were determined by RT-PCR. Representative densitometry analysis of B: TNF-α, C: IL-1β, and D: IL-6 compared with GAPDH mRNA, respectively. Data are mean ± S.E.M. (n = 3) for three independent experiments. <sup>#</sup>P<0.001 compared with the control group; **P<0.01 and ***P<0.001 compared with the LPS-treated group by One-Way analysis of variance (ANOVA) followed by <i>Bonferroni’s</i> multiple comparison test.</p>", "links"=>[], "tags"=>["glaucocalyxin-a", "pro-inflammatory", "cytokines", "necrosis", "il-6", "lipopolysaccharide", "bv-2"], "article_id"=>166853, "categories"=>["Biochemistry", "Neuroscience", "Pharmacology", "Immunology"], "users"=>["Byung-Wook Kim", "Sushruta Koppula", "Seong-Su Hong", "Sae-Bom Jeon", "Ji-Hye Kwon", "Bang-Yeon Hwang", "Eun-Jung Park", "Dong-Kug Choi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055792.g005", "stats"=>{"downloads"=>2, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_glaucocalyxin_A_GLA_on_pro_inflammatory_cytokines_such_as_tumor_necrosis_factor_945_TNF_945_interleukin_1_946_IL_1_946_and_IL_6_in_lipopolysaccharide_LPS_stimulated_BV_2_microglia_/166853", "title"=>"Effect of glaucocalyxin-A (GLA) on pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 in lipopolysaccharide (LPS)-stimulated BV-2 microglia.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:03:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/495742"], "description"=>"<p>A: Isolation procedure. B: Chemical structure of GLA.</p>", "links"=>[], "tags"=>["glaucocalyxin-a"], "article_id"=>166249, "categories"=>["Biochemistry", "Neuroscience", "Pharmacology", "Immunology"], "users"=>["Byung-Wook Kim", "Sushruta Koppula", "Seong-Su Hong", "Sae-Bom Jeon", "Ji-Hye Kwon", "Bang-Yeon Hwang", "Eun-Jung Park", "Dong-Kug Choi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055792.g001", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Isolation_and_identification_of_glaucocalyxin_A_GLA_/166249", "title"=>"Isolation and identification of glaucocalyxin-A (GLA).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 15:59:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/496221"], "description"=>"<p>Cells were pre-treated with the indicated concentrations of GLA for 1 h before incubating with LPS (25 ng/mL or 100 ng/mL) for 18 h A: Primary Microglia, B: BV-2 cells. Results are expressed as a ratio of COX-2 to β-actin. Representative quantification data was shown in the lower panel. C: Cells were pre-treated with the indicated concentrations of GLA for 1 h and then stimulated with LPS (100 ng/mL) for 6 h. Total RNA was prepared and COX-2 mRNA level was determined by RT-PCR. Results are expressed as the ratio of COX-2 to GAPDH. Quantification data are shown in the lower panel. D: PGE<sub>2</sub> levels were analyzed with an enzyme immunoassay kit. Absorbance was measured at 420 nm spectrophotometrically. Data are mean ± S.E.M. (n = 3) for three independent experiments. <sup>#</sup>P<0.001 compared with control group; *P<0.05 and ***P<0.001 compared with LPS-treated group by One-Way analysis of variance (ANOVA) followed by <i>Bonferroni’s</i> multiple comparison test.</p>", "links"=>[], "tags"=>["glaucocalyxin-a", "cyclooxygenase-2", "prostaglandin", "lipopolysaccharide"], "article_id"=>166730, "categories"=>["Biochemistry", "Neuroscience", "Pharmacology", "Immunology"], "users"=>["Byung-Wook Kim", "Sushruta Koppula", "Seong-Su Hong", "Sae-Bom Jeon", "Ji-Hye Kwon", "Bang-Yeon Hwang", "Eun-Jung Park", "Dong-Kug Choi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055792.g004", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_glaucocalyxin_A_GLA_on_cyclooxygenase_2_COX_2_and_prostaglandin_E_2_PGE_2_expression_in_lipopolysaccharide_LPS_stimulated_microglia_/166730", "title"=>"Effect of glaucocalyxin-A (GLA) on cyclooxygenase-2 (COX-2) and prostaglandin E<sub>2</sub> (PGE<sub>2</sub>) expression in lipopolysaccharide (LPS)-stimulated microglia.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:02:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/496579"], "description"=>"<p>Cells were treated with the indicated dose of GLA for 1 h before LPS (100 ng/mL) treatment for 30 min. A: Primary Microglia, B: BV-2 cells C: BV-2 cells were pre-treated with 20 µM PDTC for 2 h before 5 µM GLA was added for 1 h and before incubating with LPS (100 ng/mL) for 30 min. Lysates were analyzed by immunoblotting with an anti p-IκB-α/IκB-α antibody. Representative densitometric analyses was shown in the lower panels. Results are expressed as a ratio of p-IκB-α to IκB-α. D: BV-2 microglial cells were pre-treated with 20 µM PDTC for 2 h before 5 µM GLA was added and 1 h before incubating with LPS (100 ng/mL) for 24 h. Data are mean ± S.E.M. (n = 3) for three independent experiments. <sup>#</sup>P<0.001 compared with control group; *P<0.05, **P<0.01, and ***P<0.001 compared with LPS-treated group by One-Way analysis of variance (ANOVA) followed by <i>Bonferroni’s</i> multiple comparison test.</p>", "links"=>[], "tags"=>["glaucocalyxin-a", "phosphorylation", "degradation", "lipopolysaccharide", "microglia"], "article_id"=>167083, "categories"=>["Biochemistry", "Neuroscience", "Pharmacology", "Immunology"], "users"=>["Byung-Wook Kim", "Sushruta Koppula", "Seong-Su Hong", "Sae-Bom Jeon", "Ji-Hye Kwon", "Bang-Yeon Hwang", "Eun-Jung Park", "Dong-Kug Choi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055792.g007", "stats"=>{"downloads"=>2, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inhibitory_effect_of_glaucocalyxin_A_GLA_on_I_954_B_945_phosphorylation_and_degradation_in_lipopolysaccharide_LPS_stimulated_microglia_cells_/167083", "title"=>"Inhibitory effect of glaucocalyxin-A (GLA) on IκB-α phosphorylation and degradation in lipopolysaccharide (LPS)-stimulated microglia cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:04:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/495941"], "description"=>"<p>A: Rat primary microglial cells were pre-treated with the indicated concentrations (0.1 µM or 0.5 µM) of GLA for 1 h before incubating with LPS (25 ng/mL) for 24 h. Nitrite levels were measured in culture media by the Griess reaction. B: BV-2 microglia cells were pre-treated with the indicated concentrations of GLA (0.1, 1, or 5 µM) for 1 h before stimulating with LPS (100 ng/mL) for 24 h. C: Primary microglial cells were treated with GLA (0.1 µM) and LPS(25 ng/ml) for 18 h. Cells were stained with CD11b (OX-42) antibody and changes in morphology was observed using confocal microscopy (scale bar, 20 µm). Data are mean ± S.E.M. (n = 3) for three independent experiments. <sup>#</sup>P<0.001, compared with control group; *P<0.05, **P<0.01 and ***P<0.001 compared with LPS-treated group by One-Way analysis of variance (ANOVA) followed by <i>Bonferroni’s</i> multiple comparison test.</p>", "links"=>[], "tags"=>["glaucocalyxin-a", "nitric", "oxide", "lipopolysaccharide", "microglial"], "article_id"=>166444, "categories"=>["Biochemistry", "Neuroscience", "Pharmacology", "Immunology"], "users"=>["Byung-Wook Kim", "Sushruta Koppula", "Seong-Su Hong", "Sae-Bom Jeon", "Ji-Hye Kwon", "Bang-Yeon Hwang", "Eun-Jung Park", "Dong-Kug Choi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055792.g002", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_glaucocalyxin_A_GLA_on_nitric_oxide_NO_production_in_lipopolysaccharide_LPS_stimulated_microglial_cells_/166444", "title"=>"Effect of glaucocalyxin-A (GLA) on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated microglial cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:00:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/496096"], "description"=>"<p>Cells were pre-treated with the indicated concentrations of GLA for 1 h before incubating with LPS (25 ng/mL or 100 ng/mL) for 18 h A: Primary microglia, B: BV-2 microglia. Lysates were analyzed by immunoblotting with an anti-iNOS antibody. Representative quantification data were shown in the lower panel. Results are expressed as a ratio of iNOS to β-actin. C: BV-2 cells were pre-treated with the indicated concentrations of GLA for 1 h before incubating with LPS (100 ng/mL) for 6 h. The total RNA was isolated and iNOS mRNA level was determined by RT-PCR. Quantification data are shown in the lower panel. Results are expressed as a ratio of iNOS to GAPDH. D: BV-2 microglial cells were pre-treated with the indicated concentrations of GLA for 1 h before incubating with LPS (100 ng/mL) for 18 h. iNOS activity was analyzed with enzyme immunoassay kit. Data are mean ± S.E.M. (n = 3) for three independent experiments. <sup>#</sup>P<0.001 compared with control group; *P<0.05, **P<0.01 and ***P<0.001 compared with LPS-treated group by One-Way analysis of variance (ANOVA) followed by <i>Bonferroni’s</i> multiple comparison test.</p>", "links"=>[], "tags"=>["glaucocalyxin-a", "inducible", "nitric", "oxide", "synthase", "enzyme", "lipopolysaccharide"], "article_id"=>166606, "categories"=>["Biochemistry", "Neuroscience", "Pharmacology", "Immunology"], "users"=>["Byung-Wook Kim", "Sushruta Koppula", "Seong-Su Hong", "Sae-Bom Jeon", "Ji-Hye Kwon", "Bang-Yeon Hwang", "Eun-Jung Park", "Dong-Kug Choi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055792.g003", "stats"=>{"downloads"=>2, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_glaucocalyxin_A_GLA_on_inducible_nitric_oxide_synthase_iNOS_expression_and_enzyme_activity_in_lipopolysaccharide_LPS_stimulated_microglia_/166606", "title"=>"Effect of glaucocalyxin-A (GLA) on inducible nitric oxide synthase (iNOS) expression and enzyme activity in lipopolysaccharide (LPS)-stimulated microglia.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:01:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/496780"], "description"=>"<p>Cells were pre-treated with indicated doses of GLA for 1 h before LPS treatment (100 ng/mL) for 30 min. A: Primary Microglia, B: BV-2 microglial cells. Total protein was subjected to 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting using anti-p38 MAPKs. Representative densitometric analysis of the p38 bands were shown in the lower panel. Results are expressed as a ratio of p-p38 to total p38. C: Cells were pre-treated with the indicated doses of GLA for 1 h before LPS (100 ng/mL) treatment. LCN-2 mRNA levels were determined by RT-PCR. GAPDH was used as an internal control for the RT-PCR analysis. Representative quantification data was shown in the lower panel. Results are expressed as a ratio of LCN-2 to GAPDH. Data are mean ± S.E.M. (n = 3) for three independent experiments. <sup>#</sup>P<0.001 compared with control group; *P<0.05, **P<0.01, and ***P<0.001 compared with LPS-treated group by One-Way analysis of variance (ANOVA) followed by <i>Bonferroni’s</i> multiple comparison test.</p>", "links"=>[], "tags"=>["glaucocalyxin-a", "phosphorylation", "p38", "mitogen", "activated", "kinase", "lipocalin-2", "lipopolysaccharide", "microglia"], "article_id"=>167283, "categories"=>["Biochemistry", "Neuroscience", "Pharmacology", "Immunology"], "users"=>["Byung-Wook Kim", "Sushruta Koppula", "Seong-Su Hong", "Sae-Bom Jeon", "Ji-Hye Kwon", "Bang-Yeon Hwang", "Eun-Jung Park", "Dong-Kug Choi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055792.g008", "stats"=>{"downloads"=>3, "page_views"=>42, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inhibitory_effects_of_glaucocalyxin_A_GLA_on_phosphorylation_of_p38_mitogen_activated_protein_kinase_MAPK_and_expression_of_lipocalin_2_LCN_2_in_lipopolysaccharide_LPS_stimulated_microglia_cells_/167283", "title"=>"Inhibitory effects of glaucocalyxin-A (GLA) on phosphorylation of p38 mitogen activated protein kinase (MAPK) and expression of lipocalin-2 (LCN-2) in lipopolysaccharide (LPS)-stimulated microglia cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:05:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/496458"], "description"=>"<p>A: BV-2 microglia cells were seeded at a density of 5×10<sup>4</sup>cells/well on 24-well plates. Cells were stimulated with LPS (100 ng/mL) in the absence or presence of GLA (5 µM) added 1 h before stimulation. At 30 min after adding the LPS, the sub-cellular location of the NF-κB p65 subunit was determined by immunofluorescence assay. B: Cells were treated with the indicated dose of GLA 30 min before LPS (100 ng/mL) treatment. Total nuclear protein was subjected to 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting using anti-NF-κB p65. Densitometry analysis of NF-κB p65 is shown in the lower panel. Results are expressed as a ratio of NF-κB p65 to nucleolin. Data are mean ± S.E.M. (n = 3) for three independent experiments. <sup>#</sup> P<0.001 compared with control group; *P<0.05, **P<0.01, and ***P<0.001 compared with LPS-treated group by One-Way analysis of variance (ANOVA) followed by <i>Bonferroni’s</i> multiple comparison test.</p>", "links"=>[], "tags"=>["glaucocalyxin-a", "lipopolysaccharide", "bv-2"], "article_id"=>166971, "categories"=>["Biochemistry", "Neuroscience", "Pharmacology", "Immunology"], "users"=>["Byung-Wook Kim", "Sushruta Koppula", "Seong-Su Hong", "Sae-Bom Jeon", "Ji-Hye Kwon", "Bang-Yeon Hwang", "Eun-Jung Park", "Dong-Kug Choi"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055792.g006", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inhibitory_effect_of_glaucocalyxin_A_GLA_on_nuclear_factor_NF_954_B_activity_in_lipopolysaccharide_LPS_stimulated_BV_2_microglia_/166971", "title"=>"Inhibitory effect of glaucocalyxin-A (GLA) on nuclear factor (NF)-κB activity in lipopolysaccharide (LPS)-stimulated BV-2 microglia.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:03:37"}

PMC Usage Stats | Further Information

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Relative Metric

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