Systemic Simvastatin Rescues Retinal Ganglion Cells from Optic Nerve Injury Possibly through Suppression of Astroglial NF-κB Activation
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{"title"=>"Systemic simvastatin rescues retinal ganglion cells from optic nerve injury possibly through suppression of astroglial NF-κB activation", "type"=>"journal", "authors"=>[{"first_name"=>"Seita", "last_name"=>"Morishita", "scopus_author_id"=>"56548279100"}, {"first_name"=>"Hidehiro", "last_name"=>"Oku", "scopus_author_id"=>"7102887179"}, {"first_name"=>"Taeko", "last_name"=>"Horie", "scopus_author_id"=>"43361494800"}, {"first_name"=>"Masahiro", "last_name"=>"Tonari", "scopus_author_id"=>"23393787400"}, {"first_name"=>"Teruyo", "last_name"=>"Kida", "scopus_author_id"=>"7102598529"}, {"first_name"=>"Akiko", "last_name"=>"Okubo", "scopus_author_id"=>"56021514100"}, {"first_name"=>"Tetsuya", "last_name"=>"Sugiyama", "scopus_author_id"=>"7402751345"}, {"first_name"=>"Shinji", "last_name"=>"Takai", "scopus_author_id"=>"7201470915"}, {"first_name"=>"Hideaki", "last_name"=>"Hara", "scopus_author_id"=>"56799750000"}, {"first_name"=>"Tsunehiko", "last_name"=>"Ikeda", "scopus_author_id"=>"7404132784"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84894268358", "sgr"=>"84894268358", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0084387", "pmid"=>"24392131", "pui"=>"372426302"}, "id"=>"1f87b69b-8199-30b3-b39d-16f3ee6abfd8", "abstract"=>"Neuroinflammation is involved in the death of retinal ganglion cells (RGCs) after optic nerve injury. The purpose of this study was to determine whether systemic simvastatin can suppress neuroinflammation in the optic nerve and rescue RGCs after the optic nerve is crushed. Simvastatin or its vehicle was given through an osmotic minipump beginning one week prior to the crushing. Immunohistochemistry and real-time PCR were used to determine the degree of neuroinflammation on day 3 after the crushing. The density of RGCs was determined in Tuj-1 stained retinal flat mounts on day 7. The effect of simvastain on the TNF-α-induced NF-κB activation was determined in cultured optic nerve astrocytes. On day 3, CD68-positive cells, most likely microglia/macrophages, were accumulated at the crushed site. Phosphorylated NF-κB was detected in some astrocytes at the border of the lesion where the immunoreactivity to MCP-1 was intensified. There was an increase in the mRNA levels of the CD68 (11.4-fold), MCP-1 (22.6-fold), ET-1 (2.3-fold), GFAP (1.6-fold), TNF-α (7.0-fold), and iNOS (14.8-fold) genes on day 3. Systemic simvastatin significantly reduced these changes. The mean ± SD number of RGCs was 1816.3±232.6/mm(2) (n = 6) in the sham controls which was significantly reduced to 831.4±202.5/mm(2) (n = 9) on day 7 after the optic nerve was crushed. This reduction was significantly suppressed to 1169.2±201.3/mm(2) (P = 0.01, Scheffe; n = 9) after systemic simvastatin. Simvastatin (1.0 µM) significantly reduced the TNF-α-induced NF-κB activation in cultured optic nerve astrocytes. We conclude that systemic simvastatin can reduce the death of RGCs induced by crushing the optic nerve possibly by suppressing astroglial NF-κB activation.", "link"=>"http://www.mendeley.com/research/systemic-simvastatin-rescues-retinal-ganglion-cells-optic-nerve-injury-possibly-through-suppression", "reader_count"=>11, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Student > Ph. D. Student"=>5, "Student > Postgraduate"=>2, "Student > Master"=>2, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Student > Ph. D. Student"=>5, "Student > Postgraduate"=>2, "Student > Master"=>2, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>4, "Medicine and Dentistry"=>4, "Neuroscience"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Neuroscience"=>{"Neuroscience"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>4}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}}, "reader_count_by_country"=>{"Brazil"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1337101"], "description"=>"<p>RNA was extracted from 4(placebo). Systemic simvastatin suppresses the increase significantly (n = 6 to 8 for each condition; 22 rats were used). Data are shown as the fold changes (mean ± SD) to the sham control in the mRNA expressions.</p>", "links"=>[], "tags"=>["immunology", "immunity", "inflammation", "microbiology", "Model organisms", "Animal models", "rat", "Molecular cell biology", "Cell death", "neuroscience", "Neurobiology of disease and regeneration", "Drugs and devices", "Drug research and development", "neurology", "Neuro-ophthalmology", "Neuropharmacology", "ophthalmology", "glaucoma", "mrna", "genes", "optic", "crushing"], "article_id"=>892877, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Seita Morishita", "Hidehiro Oku", "Taeko Horie", "Masahiro Tonari", "Teruyo Kida", "Akiko Okubo", "Tetsuya Sugiyama", "Shinji Takai", "Hideaki Hara", "Tsunehiko Ikeda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084387.g005", "stats"=>{"downloads"=>2, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Changes_in_the_mRNA_levels_of_the_CD68_MCP_1_ET_1_GFAP_TNF_945_and_iNOS_genes_in_the_optic_nerve_on_day_3_after_crushing_the_optic_nerves_/892877", "title"=>"Changes in the mRNA levels of the <i>CD68</i>, <i>MCP-1</i>, <i>ET-1</i>, <i>GFAP</i>, <i>TNF-α</i>, and <i>iNOS</i> genes in the optic nerve on day 3 after crushing the optic nerves.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-02 03:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1337091"], "description"=>"<p>Representative photographs from 3 independent samples for each condition are presented. In each picture, 45 images were stacked using Z-scan at 0.5 µm intervals. CD68-positive cells are not visible in the sham control (A), while many CD68-positive cells (green) are present at the crushed site (arrows) in the optic nerve with placebo treatment (B). Immunoreactivity to GFAP (red) was relatively weak at the crush site, and it was intensified in the area surrounding the lesion. This relationship suggests that chemoattractant molecules are expressed in reactive astrocytes at the border of the crush site. The accumulation of CD68-positive cells and immunoreactivity to GFAP is reduced by systemic simvastatin treatment (C). CD68 staining: mouse monoclonal anti-CD68 (primary) and Alexa 488-conjugated goat anti-mouse IgG (secondary antibodies); GFAP staining: rabbit polyclonal anti-GFAP (primary) and Alexa 594-conjugated goat anti-rabbit IgG (secondary antibodies). Bar = 100 µm. A total of 9 rats were used in these analyses.</p>", "links"=>[], "tags"=>["immunology", "immunity", "inflammation", "microbiology", "Model organisms", "Animal models", "rat", "Molecular cell biology", "Cell death", "neuroscience", "Neurobiology of disease and regeneration", "Drugs and devices", "Drug research and development", "neurology", "Neuro-ophthalmology", "Neuropharmacology", "ophthalmology", "glaucoma", "cd68", "gfap", "crushed", "optic"], "article_id"=>892867, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Seita Morishita", "Hidehiro Oku", "Taeko Horie", "Masahiro Tonari", "Teruyo Kida", "Akiko Okubo", "Tetsuya Sugiyama", "Shinji Takai", "Hideaki Hara", "Tsunehiko Ikeda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084387.g002", "stats"=>{"downloads"=>8, "page_views"=>348, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemistry_for_CD68_and_GFAP_expression_in_the_crushed_optic_nerve_on_day_3_/892867", "title"=>"Immunohistochemistry for CD68 and GFAP expression in the crushed optic nerve on day 3.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-02 03:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1337103"], "description"=>"<p>A. Characterization of cultured astrocytes isolated from optic nerves. Left panels; GFAP immunostaining of cultured astrocytes with DAPI nuclear counter staining. All cells are stained positively with antibody to GFAP (green). These cells are also positively stained with TNFR1 antibody (red). Bar = 100 µm. Right panel; FACS analysis of GFAP expression on our cultured astrocytes. Over 96% of the cells express GFAP. B. Representative western blot analysis of phosphorylated NF- κB p65 and total NF- κB p65. Tubulin was used as an internal control. C. Phosphorylation of NF- κB was quantified by the ratio of phosphorylated NF- κB to the total NF- κB levels. Data are shown as fold changes (mean ± SD, n = 4 for each) to the control. TNF-α induced a 1.4-fold increase of phosphorylated NF-κB from the control level, which was significantly (<i>P</i> = 0.001, Scheffe) suppressed by simvastatin. Addition of mevalonate (100 µM) significantly (<i>P</i> = 0.04, Scheffe) restored the increase. Incubation with NF- κB activation inhibitor, QNZ completely suppressed the TNF-α-induced phosphorylation of NF-κB. D. Representative EMSA analysis of NF-κB DNA binding activity. NF-κB DNA–binding activity in the nuclei of optic nerve astrocytes in the control medium (Lane 1), exposed to TNF-α (50 ng/ml) (Lane 2), exposed to TNF-α (50 ng/ml) and simvastatin (1.0 µM) (Lane 3), incubated with simvastatin (1.0 µM) alone (lane 4), NF-κB DNA–binding activity by competition EMSA with a 100-fold excess of cold NF-κB oligonucleotides (Lane 5). E. Quantification of NF-κB DNA–binding activity in the nuclei of optic nerve astrocytes cultured under different conditions. Fold changes (mean ± SD, n = 4 for each) from the control are plotted. Exposed to TNF-α (50 ng/ml) caused a 3.8 fold increase of NF-κB DNA–binding activity, which was significantly (<i>P</i><0.001, Scheffe) suppressed by co-incubation with simvastatin (1.0 µM).</p>", "links"=>[], "tags"=>["immunology", "immunity", "inflammation", "microbiology", "Model organisms", "Animal models", "rat", "Molecular cell biology", "Cell death", "neuroscience", "Neurobiology of disease and regeneration", "Drugs and devices", "Drug research and development", "neurology", "Neuro-ophthalmology", "Neuropharmacology", "ophthalmology", "glaucoma", "simvastatin", "activation", "optic"], "article_id"=>892879, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Seita Morishita", "Hidehiro Oku", "Taeko Horie", "Masahiro Tonari", "Teruyo Kida", "Akiko Okubo", "Tetsuya Sugiyama", "Shinji Takai", "Hideaki Hara", "Tsunehiko Ikeda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084387.g006", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_simvastatin_on_the_TNF_945_induced_NF_954_B_activation_of_optic_nerve_astrocytes_/892879", "title"=>"Effects of simvastatin on the TNF-α-induced NF-κB activation of optic nerve astrocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-02 03:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1337099"], "description"=>"<p>Representative photographs from 3 independent samples for each condition are presented. A. Phosphorylated NF-κB p65 can be detected at the border of the crushed site (arrowheads, crush placebo). Systemic simvastatin appears to decrease the number of immunopositive cells to phosphorylated NF-κB p65 (arrowheads, crush statin). There are no apparent positive cells in the sham control (sham). Images of negative control without primary antibodies were prepared from animals that underwent crushing of the optic nerve. Arrows indicate crushed site. Bar = 100 µm. B. Marged images of phosphorylated NF-κB p65, DAPI and GFAP at the margin of the crushed site (crush, placebo). Phosphorylation of NF-κB, which indicates NF-κB activation, can be detected in some nuclei (left panel). Immunoreactivity to phosphorylated NF-κB is colocalized with some GFAP positive cells at the border of the crushed site (right panel). Phosphorylated NF-κB p65 staining: rabbit polyclonal anti- phosphorylated NF-κB p65 (primary) and alexa 594-conjugated goat anti-rabbit IgG (secondary antibodies); GFAP staining: mouse monoclonal anti-GFAP (primary) and Alexa 488-conjugated goat anti-mouse IgG (secondary antibodies). Bar = 100 µm. A total of 12 rats were used in these analyses.</p>", "links"=>[], "tags"=>["immunology", "immunity", "inflammation", "microbiology", "Model organisms", "Animal models", "rat", "Molecular cell biology", "Cell death", "neuroscience", "Neurobiology of disease and regeneration", "Drugs and devices", "Drug research and development", "neurology", "Neuro-ophthalmology", "Neuropharmacology", "ophthalmology", "glaucoma", "phosphorylated", "p65", "gfap", "crushed", "optic", "sham"], "article_id"=>892875, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Seita Morishita", "Hidehiro Oku", "Taeko Horie", "Masahiro Tonari", "Teruyo Kida", "Akiko Okubo", "Tetsuya Sugiyama", "Shinji Takai", "Hideaki Hara", "Tsunehiko Ikeda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084387.g004", "stats"=>{"downloads"=>5, "page_views"=>37, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemistry_for_phosphorylated_NF_954_B_p65_and_GFAP_at_the_margin_of_the_crushed_site_of_the_optic_nerve_from_sham_control_and_from_experimental_animals_/892875", "title"=>"Immunohistochemistry for phosphorylated NF-κB p65 and GFAP at the margin of the crushed site of the optic nerve from sham control and from experimental animals.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-02 03:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1337092"], "description"=>"<p>Representative photographs from 3 independent samples for each condition are presented. A. Consistent with the findings shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084387#pone-0084387-g002\" target=\"_blank\">Figure 2</a>, immunoreactivity to GFAP (green) was weak at the crush site, and immunoreactivity to MCP-1 (red) is also weak (crush placebo). Crushing the optic nerve intensified the immunoreactivity to MCP-1 in the area surrounding the crush site, where immunoreactivity to GFAP was increased (crush placebo) compared to the sham control. Systemic simvastatin decreases the immunoreactivities to the MCP-1 and GFAP in the area surrounding the crush site (crush statin). Images of negative control without primary antibodies were prepared from animals that underwent crushing the optic nerve. Arrows indicate crushed sites. Bar = 100 µm. B. Confocal images at the margin of the crushed site from vehicle-treated rats (square area). Immunoreactivity to MCP-1 is well co-localized with that to GFAP. MCP-1 staining: rabbit polyclonal anti-MCP-1 (primary) and Alexa 594-conjugated goat anti-rabbit IgG (secondary antibodies); GFAP staining: mouse monoclonal anti-GFAP (primary) and Alexa 488-conjugated goat anti-mouse IgG (secondary antibodies). Bar = 100 µm. A total of 12 rats were used in these analyses.</p>", "links"=>[], "tags"=>["immunology", "immunity", "inflammation", "microbiology", "Model organisms", "Animal models", "rat", "Molecular cell biology", "Cell death", "neuroscience", "Neurobiology of disease and regeneration", "Drugs and devices", "Drug research and development", "neurology", "Neuro-ophthalmology", "Neuropharmacology", "ophthalmology", "glaucoma", "mcp-1", "gfap", "optic", "sham", "animals", "crushing"], "article_id"=>892868, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Seita Morishita", "Hidehiro Oku", "Taeko Horie", "Masahiro Tonari", "Teruyo Kida", "Akiko Okubo", "Tetsuya Sugiyama", "Shinji Takai", "Hideaki Hara", "Tsunehiko Ikeda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084387.g003", "stats"=>{"downloads"=>14, "page_views"=>405, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunohistochemistry_for_MCP_1_and_GFAP_in_the_optic_nerve_from_a_sham_control_and_from_experimental_animals_after_crushing_the_optic_nerves_/892868", "title"=>"Immunohistochemistry for MCP-1 and GFAP in the optic nerve from a sham control and from experimental animals after crushing the optic nerves.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-02 03:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1337106", "https://ndownloader.figshare.com/files/1337107"], "description"=>"<div><p>Neuroinflammation is involved in the death of retinal ganglion cells (RGCs) after optic nerve injury. The purpose of this study was to determine whether systemic simvastatin can suppress neuroinflammation in the optic nerve and rescue RGCs after the optic nerve is crushed. Simvastatin or its vehicle was given through an osmotic minipump beginning one week prior to the crushing. Immunohistochemistry and real-time PCR were used to determine the degree of neuroinflammation on day 3 after the crushing. The density of RGCs was determined in Tuj-1 stained retinal flat mounts on day 7. The effect of simvastain on the TNF-α-induced NF-κB activation was determined in cultured optic nerve astrocytes. On day 3, CD68-positive cells, most likely microglia/macrophages, were accumulated at the crushed site. Phosphorylated NF-κB was detected in some astrocytes at the border of the lesion where the immunoreactivity to MCP-1 was intensified. There was an increase in the mRNA levels of the <i>CD68</i> (11.4-fold), <i>MCP-1</i> (22.6-fold), <i>ET-1</i> (2.3-fold), <i>GFAP</i> (1.6-fold), <i>TNF-α</i> (7.0-fold), and <i>iNOS</i> (14.8-fold) genes on day 3. Systemic simvastatin significantly reduced these changes. The mean ± SD number of RGCs was 1816.3±232.6/mm<sup>2</sup> (n = 6) in the sham controls which was significantly reduced to 831.4±202.5/mm<sup>2</sup> (n = 9) on day 7 after the optic nerve was crushed. This reduction was significantly suppressed to 1169.2±201.3/mm<sup>2</sup> (<i>P</i> = 0.01, Scheffe; n = 9) after systemic simvastatin. Simvastatin (1.0 µM) significantly reduced the TNF-α-induced NF-κB activation in cultured optic nerve astrocytes. We conclude that systemic simvastatin can reduce the death of RGCs induced by crushing the optic nerve possibly by suppressing astroglial NF-κB activation.</p></div>", "links"=>[], "tags"=>["immunology", "immunity", "inflammation", "microbiology", "Model organisms", "Animal models", "rat", "Molecular cell biology", "Cell death", "neuroscience", "Neurobiology of disease and regeneration", "Drugs and devices", "Drug research and development", "neurology", "Neuro-ophthalmology", "Neuropharmacology", "ophthalmology", "glaucoma", "simvastatin", "rescues", "retinal", "ganglion", "cells", "optic", "suppression", "astroglial"], "article_id"=>892882, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Seita Morishita", "Hidehiro Oku", "Taeko Horie", "Masahiro Tonari", "Teruyo Kida", "Akiko Okubo", "Tetsuya Sugiyama", "Shinji Takai", "Hideaki Hara", "Tsunehiko Ikeda"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0084387.s001", "https://dx.doi.org/10.1371/journal.pone.0084387.s002"], "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Systemic_Simvastatin_Rescues_Retinal_Ganglion_Cells_from_Optic_Nerve_Injury_Possibly_through_Suppression_of_Astroglial_NF_954_B_Activation_/892882", "title"=>"Systemic Simvastatin Rescues Retinal Ganglion Cells from Optic Nerve Injury Possibly through Suppression of Astroglial NF-κB Activation", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-01-02 03:22:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/1337087"], "description"=>"<p>A. Retinas from sham control (left), crushed optic nerves treated with placebo (middle), crushed optic nerves treated with simvastatin (right panel). Pictures were taken 1.5 mm from the optic disc margin. Bar = 100 µm. B. The density of retinal ganglion cells (RGCs/mm<sup>2</sup>) is quantified. Systemic simvastatin had a significant (<i>P</i> = 0.01, Scheffe) protective effect on the RGCs.</p>", "links"=>[], "tags"=>["immunology", "immunity", "inflammation", "microbiology", "Model organisms", "Animal models", "rat", "Molecular cell biology", "Cell death", "neuroscience", "Neurobiology of disease and regeneration", "Drugs and devices", "Drug research and development", "neurology", "Neuro-ophthalmology", "Neuropharmacology", "ophthalmology", "glaucoma", "photomicrographs", "mounted", "retinas", "stained", "alexa", "488-conjugated", "tuj-1"], "article_id"=>892863, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Seita Morishita", "Hidehiro Oku", "Taeko Horie", "Masahiro Tonari", "Teruyo Kida", "Akiko Okubo", "Tetsuya Sugiyama", "Shinji Takai", "Hideaki Hara", "Tsunehiko Ikeda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084387.g001", "stats"=>{"downloads"=>12, "page_views"=>333, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Representative_photomicrographs_of_flat_mounted_retinas_stained_with_Alexa_488_conjugated_Tuj_1_antibody_/892863", "title"=>"Representative photomicrographs of flat mounted retinas stained with Alexa 488-conjugated Tuj-1 antibody.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-02 03:22:49"}

PMC Usage Stats | Further Information

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Relative Metric

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