Journal: Neuropsychopharmacology

Article Title: The Modulatory Role of the Lateral Septum on Neuroendocrine and Behavioral Stress Responses

doi: 10.1038/npp.2010.213

Figure Lengend Snippet: Effects of intraseptal administration of the glucocorticoid receptor antagonist mifepristone (20 or 100 ng/side; black arrow) on neuroendocrine and behavioral stress response. Administration of mifepristone had no effect on either basal or stress-induced increase of plasma ACTH (a) or corticosterone (CORT, b) levels because there was no significant difference between mifepristone-treated animals and controls. (c) Mifepristone-injected animals showed an increased active coping style indicated by an enhancement of struggling and swimming and a reduction of floating behavior during the forced swim exposure. Data are expressed as means+SEM. ** p <0.01 vs basal values; ++ p <0.001 vs vehicle-treated controls (Newman–Keuls post hoc test or Student's t -test).

Article Snippet: Injection cannulae were connected to a syringe mounted on a microinfusion pump (TSE-Systems, Bad Homburg, Germany) and filled either with artificial cerebrospinal fluid (aCSF; pH 7.2; 140 mM NaCl, 3.0 mM KCl, 1.25 mM CaCl 2 , 1.0 mM MgCl 2 , 1.2 mM Na 2 HPO 4 , 0.3 mM NaH 2 PO 4 , and 3.0 mM glucose) or aCSF containing the corticosteroid receptor antagonists mifepristone or RU-28318 (30 and 150 μM; Tocris Bioscience, Bristol, UK).

Techniques: Clinical Proteomics, Injection

Journal: Blood

Article Title: Rapid immunosuppressive effects of glucocorticoids mediated through Lck and Fyn.

doi: 10.1182/blood-2004-12-4790

Figure Lengend Snippet: Figure 6. DEX-induced inhibition of Lck and Fyn kinase activities is GR dependent. (A) GR-negative Jurkat cells (confirmed by Western blot analysis; not shown) were pretreated with DEX for 10 minutes and incubated with anti-CD3, anti-CD28 mAbs for 15 minutes. Whole-cell lysates were used for Western blot analysis. Because there are no Abs commercially available targeting phospho-Lck associated with increased activity, phospho-ZAP70 expression (a downstream target of Lck) was analyzed. DEX treatment did not affect phospho-Fyn expression, nor did it affect phospho-ZAP70 protein levels in GR-negative Jurkat cells. Three experi- ments were performed, and similar outcomes were obtained. (B) CD4 T cells were incubated for 1 hour with a GR mimetic (RU486; 50 M), which acts as a GR agonist at this concentration. Subsequently, cells were treated with DEX (10 minutes) and activated with anti-CD3, anti-CD28 Abs (15 minutes). Lck and Fyn immunoprecipi- tates were subjected to in vitro kinase assay, and phospho-SAM68 expression was studied on Western blot using PY20. Decreased phospo-SAM68 expression was seen in Lck and Fyn immunoprecipitates prepared from activated cells treated with DEX, and cells treated with RU486 together with DEX. Incubation with RU486 alone (at this high/agonistic concentration) also resulted in impaired SAM68 phosphoryla- tion. DEX indicates dexamethasone; IB, immunoblotting; IP, immunoprecipitation.

Article Snippet: RU486 (mifepristone) was obtained from LKT Laboratories (Minnesota, MN).

Techniques: Inhibition, Western Blot, Incubation, Activity Assay, Expressing, Concentration Assay, In Vitro, Kinase Assay, Immunoprecipitation

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Experimental groups

Article Snippet: C57BL/6 mice were randomly divided into five groups: (1) Control group, which underwent the surgical procedures but without TBI; (2) TBI group, which received saline for 3 days; (3) TBI + DEX group (Sigma–Aldrich, D4902), which was administered an intraperitoneal injection once per day for 3 days; (4) DEX+RU486 (Abcam, Cambridge, UK, Cat# ab120356), which received a GR antagonist, RU486, twice a day for 1 day before TBI and 3 days after surgery; administration of DEX was consistent with group (3); and (5) RU486 + ruxolitinib group (TargetMol, Wellesley Hills, MA, USA, Cat# 1,092,939-17-7), which received RU486 as in group (4) , while the Janus kinase (JAK) inhibitor, ruxolitinib, was administered twice daily for three days post-TBI (Lescoat et al., 2020; Chen et al., 2021).

Techniques: Control, Saline, Western Blot, TUNEL Assay, Staining, Immunofluorescence

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Reduction in M2 microglial expression by DEX is regulated by GR/JAK1/STAT3 pathway activation. (A) qRT-PCR detected M2 markers, Arg1, JAK1, STAT3, Bcl2, and Myc. (B) Western blotting measured activation levels of glucocorticoid receptors (GR) (83 kDa), and expression levels of Arg1 (35 kDa), JAK1 (133 kDa), p-STAT3 (98 kDa), Bcl2 (26 kDa), c-Myc (57 kDa) and GAPDH (36 kDa). Data are expressed as mean ± SD, n = 3. * P < 0.05, vs . Control; # P < 0.05, vs. TBI (one-way analysis of variance with the least significant difference test). DEX: Dexamethasone; TBI: traumatic brain injury.

Article Snippet: C57BL/6 mice were randomly divided into five groups: (1) Control group, which underwent the surgical procedures but without TBI; (2) TBI group, which received saline for 3 days; (3) TBI + DEX group (Sigma–Aldrich, D4902), which was administered an intraperitoneal injection once per day for 3 days; (4) DEX+RU486 (Abcam, Cambridge, UK, Cat# ab120356), which received a GR antagonist, RU486, twice a day for 1 day before TBI and 3 days after surgery; administration of DEX was consistent with group (3); and (5) RU486 + ruxolitinib group (TargetMol, Wellesley Hills, MA, USA, Cat# 1,092,939-17-7), which received RU486 as in group (4) , while the Janus kinase (JAK) inhibitor, ruxolitinib, was administered twice daily for three days post-TBI (Lescoat et al., 2020; Chen et al., 2021).

Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Western Blot, Control, Analysis, Gene Expression, Real-time Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Expression patterns of Arg1 following TBI. (A, B) Representative immunofluorescence staining (A) and quantification (B) showing expression levels of Arg1 in the peri-injured cortex and hippocampus after TBI. Inhibition of glucocorticoid receptor activation reversed M2 microglial marker expression. Red represents Arg1, and blue represents DAPI. Scale bars: 20 μm. Data are expressed as mean ± SD, n = 3. * P < 0.05, vs . Control; # P < 0.05, vs. TBI (one-way analysis of variance with the least significant difference test). DAPI: 4′,6-Diamidino-2-phenylindole; DEX: dexamethasone; TBI: traumatic brain injury.

Article Snippet: C57BL/6 mice were randomly divided into five groups: (1) Control group, which underwent the surgical procedures but without TBI; (2) TBI group, which received saline for 3 days; (3) TBI + DEX group (Sigma–Aldrich, D4902), which was administered an intraperitoneal injection once per day for 3 days; (4) DEX+RU486 (Abcam, Cambridge, UK, Cat# ab120356), which received a GR antagonist, RU486, twice a day for 1 day before TBI and 3 days after surgery; administration of DEX was consistent with group (3); and (5) RU486 + ruxolitinib group (TargetMol, Wellesley Hills, MA, USA, Cat# 1,092,939-17-7), which received RU486 as in group (4) , while the Janus kinase (JAK) inhibitor, ruxolitinib, was administered twice daily for three days post-TBI (Lescoat et al., 2020; Chen et al., 2021).

Techniques: Expressing, Immunofluorescence, Staining, Inhibition, Activation Assay, Marker, Control, Analysis

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: RU486 and ruxolitinib treatment decreases DEX-induced apoptosis following TBI. (A, B) Representative photomicrographs (A) and quantification (B) of terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining. Apoptotic cells were measured in the ipsilateral hippocampus and cortex surrounding the injury site. Green represents TUNEL-positive cells. Blue represents DAPI. Scale bars: 20 μm. Data are expressed as mean ± SD, n = 3. * P < 0.05, vs . Control; # P < 0.05, vs. traumatic brain injury (TBI) (one-way analysis of variance with the least significant difference test). DAPI: 4′,6-Diamidino-2-phenylindole; DEX: dexamethasone; TBI: traumatic brain injury.

Article Snippet: C57BL/6 mice were randomly divided into five groups: (1) Control group, which underwent the surgical procedures but without TBI; (2) TBI group, which received saline for 3 days; (3) TBI + DEX group (Sigma–Aldrich, D4902), which was administered an intraperitoneal injection once per day for 3 days; (4) DEX+RU486 (Abcam, Cambridge, UK, Cat# ab120356), which received a GR antagonist, RU486, twice a day for 1 day before TBI and 3 days after surgery; administration of DEX was consistent with group (3); and (5) RU486 + ruxolitinib group (TargetMol, Wellesley Hills, MA, USA, Cat# 1,092,939-17-7), which received RU486 as in group (4) , while the Janus kinase (JAK) inhibitor, ruxolitinib, was administered twice daily for three days post-TBI (Lescoat et al., 2020; Chen et al., 2021).

Techniques: TUNEL Assay, Staining, Control, Analysis, Immunofluorescence

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Inhibition of GR attenuates neuronal death. (A) Representative image of Nissl staining in the peri-injured cortex and hippocampus. (B) Quantification analysis showed that DEX induced a significant decrease in Nissl-positive cells post-TBI. RU486 application alleviated neuronal loss. Scale bars: 20 μm. Data are expressed as mean ± SD, n = 3. * P < 0.05, vs . Control; # P < 0.05, vs. TBI (one-way analysis of variance with the least significant difference test). DEX: Dexamethasone; TBI: traumatic brain injury.

Article Snippet: C57BL/6 mice were randomly divided into five groups: (1) Control group, which underwent the surgical procedures but without TBI; (2) TBI group, which received saline for 3 days; (3) TBI + DEX group (Sigma–Aldrich, D4902), which was administered an intraperitoneal injection once per day for 3 days; (4) DEX+RU486 (Abcam, Cambridge, UK, Cat# ab120356), which received a GR antagonist, RU486, twice a day for 1 day before TBI and 3 days after surgery; administration of DEX was consistent with group (3); and (5) RU486 + ruxolitinib group (TargetMol, Wellesley Hills, MA, USA, Cat# 1,092,939-17-7), which received RU486 as in group (4) , while the Janus kinase (JAK) inhibitor, ruxolitinib, was administered twice daily for three days post-TBI (Lescoat et al., 2020; Chen et al., 2021).

Techniques: Inhibition, Staining, Analysis, Control

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Golgi-stained neurons show an effect of DEX on dendritic spine number. (A) Representative neurons in the cortex and hippocampus are shown across the different groups. Scale bars: 10 μm. (B) Data analysis showed that neuronal complexity differed across several groups following TBI. Data are expressed as mean ± SD, n = 3. * P < 0.05, vs . Control; # P < 0.05, vs. TBI (one-way analysis of variance with the least significant difference test). DEX: Dexamethasone; TBI: traumatic brain injury.

Article Snippet: C57BL/6 mice were randomly divided into five groups: (1) Control group, which underwent the surgical procedures but without TBI; (2) TBI group, which received saline for 3 days; (3) TBI + DEX group (Sigma–Aldrich, D4902), which was administered an intraperitoneal injection once per day for 3 days; (4) DEX+RU486 (Abcam, Cambridge, UK, Cat# ab120356), which received a GR antagonist, RU486, twice a day for 1 day before TBI and 3 days after surgery; administration of DEX was consistent with group (3); and (5) RU486 + ruxolitinib group (TargetMol, Wellesley Hills, MA, USA, Cat# 1,092,939-17-7), which received RU486 as in group (4) , while the Janus kinase (JAK) inhibitor, ruxolitinib, was administered twice daily for three days post-TBI (Lescoat et al., 2020; Chen et al., 2021).

Techniques: Staining, Analysis, Control

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: The polarization level of M2 microglia is associated with spatial memory after TBI. (A) Escape latency of mice in the Morris water maze probe test before TBI. (B) Measurement of the percentage time spent in the target quadrant before and after TBI. Data are expressed as mean ± SD, n = 3. * P < 0.05, vs . Control; # P < 0.05, vs . TBI (one-way analysis of variance with the least significant difference test). DEX: Dexamethasone; MWM: Morris water maze; TBI: traumatic brain injury.

Article Snippet: C57BL/6 mice were randomly divided into five groups: (1) Control group, which underwent the surgical procedures but without TBI; (2) TBI group, which received saline for 3 days; (3) TBI + DEX group (Sigma–Aldrich, D4902), which was administered an intraperitoneal injection once per day for 3 days; (4) DEX+RU486 (Abcam, Cambridge, UK, Cat# ab120356), which received a GR antagonist, RU486, twice a day for 1 day before TBI and 3 days after surgery; administration of DEX was consistent with group (3); and (5) RU486 + ruxolitinib group (TargetMol, Wellesley Hills, MA, USA, Cat# 1,092,939-17-7), which received RU486 as in group (4) , while the Janus kinase (JAK) inhibitor, ruxolitinib, was administered twice daily for three days post-TBI (Lescoat et al., 2020; Chen et al., 2021).

Techniques: Control, Analysis