Journal: CNS Neuroscience & Therapeutics

Article Title: Calcitonin gene‐related peptide induces the histone H3 lysine 9 acetylation in astrocytes associated with neuroinflammation in rats with neuropathic pain

doi: 10.1111/cns.13720

Figure Lengend Snippet: Intrathecal CGRP antagonist and HAT inhibitor administration prevent the pain hypersensitivity and attenuate increased levels of H3K9ac in the spinal dorsal horn induced by CCI. (A) Shows the mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) during the 10‐day observation period in rats treated with daily intrathecal injection of either 1 μM CGRP, 20 μM AA (HAT inhibitor), 2 μM CGRP8‐37, or vehicle in 10 μl for 9 days. All values are expressed as the means ± SEMs ( n = 6). (B–E) Western blot analyses for H3K9ac protein levels in the dorsal quadrant of L4‐L5 spinal segment ipsilateral to the operation side with CCI surgery for 3, 5, 7, and 10 days, respectively. Data were obtained from animals treated with daily intrathecal injection of either 1 μM CGRP, 20 μM AA, 2 μM CGRP8‐37, or vehicle in 10 μl for 2, 4, 6, and 9 days, respectively. The mean optic densities of the proteins were calculated by normalizing to GAPDH. All values are expressed as the means ± SEMs ( n = 4).* p < 0.05 versus sham groups; # p < 0.05 versus CCI alone groups. (F) Double‐staining immunofluorescent images showing the expression of H3K9ac (green) in astrocytes (GFAP, red) of L4‐L5 spinal dorsal horn ipsilateral to the operation side in the sham, sham + AA, sham + CGRP, CCI, CCI + AA, and CCI + CGRP8‐37 groups on day 10 after surgery, respectively. Scale bar 100 μm. Graph showing the percentages of GFAP/H3K9ac double‐labeled cells in L4‐L5 spinal dorsal horn of the sham, sham + AA, sham + CGRP, CCI, CCI + AA, and CCI + CGRP8‐37 groups, respectively. Data are presented as the mean ± SEM ( n = 6). * p < 0.05 versus sham groups; # p < 0.05 versus CCI alone groups

Article Snippet: The CGRP (1 μM, Tocris Bioscience), anacardic acid (AA, HAT inhibitor, 20 μM, Abcam), CGRP8‐37 (CGRP antagonist, 2 μM, MCE), or vehicle in a volume of 10 μl was injected into the spinal lumbar enlargement region through the intrathecal catheter, followed by 20 μl of saline to flush.

Techniques: Injection, Western Blot, Double Staining, Expressing, Labeling

Journal: CNS Neuroscience & Therapeutics

Article Title: Calcitonin gene‐related peptide induces the histone H3 lysine 9 acetylation in astrocytes associated with neuroinflammation in rats with neuropathic pain

doi: 10.1111/cns.13720

Figure Lengend Snippet: CGRP evokes increases in the expression of H3K9ac in astroglial cells by HATs. (A) The expressions of GFAP (a marker of astrocytes, red) and its co‐localization with CRLR (green), RAMP1 (green), or CRCP (green) staining in cultured astroglial cells (C6). Scale bar 40 μm. (B) Western blot analysis of H3K9ac expression in astrocytic cell line (C6) with treatment of CGRP at 0, 1, 2, 4, 6, and 12 h, respectively. (C) Western blot analyses for H3K9ac protein levels in astroglial cells (C6) with co‐treatment of CGRP (1 μM) and AA (20 μM) for 4 h. (D) Western blot analysis of GFAP expression in astroglial cell line (C6) with treatment of CGRP at 0, 1, 2, 4, and 6 h, respectively. The mean optic densities of the proteins were calculated by normalizing to GAPDH. All values are expressed as the means ± SEMs ( n = 4).* p < 0.05 versus controls; # p < 0.05 versus CGRP only groups. (E) Shown is an example of astrocytic cell growth curves by RTCA. RTCA was performed to evaluate the proliferation and viability of astroglial cells with continuous treatment of CGRP and co‐treatment of AA or CGRP8‐37

Article Snippet: The CGRP (1 μM, Tocris Bioscience), anacardic acid (AA, HAT inhibitor, 20 μM, Abcam), CGRP8‐37 (CGRP antagonist, 2 μM, MCE), or vehicle in a volume of 10 μl was injected into the spinal lumbar enlargement region through the intrathecal catheter, followed by 20 μl of saline to flush.

Techniques: Expressing, Marker, Staining, Cell Culture, Western Blot

Journal: CNS Neuroscience & Therapeutics

Article Title: Calcitonin gene‐related peptide induces the histone H3 lysine 9 acetylation in astrocytes associated with neuroinflammation in rats with neuropathic pain

doi: 10.1111/cns.13720

Figure Lengend Snippet: CGRP altered the gene expression in astroglial cells associated with astrocyte activation. (A) Quantitative RT‐PCR analysis for differences in expression levels of H3K9ac specific target genes between CGRP‐treated astroglial cells and controls in the subset of genes gaining or losing H3K9ac on their promoters. Results were calculated by normalizing to GAPDH in the same sample with the ΔCt method. Changes in relative levels of gene mRNAs expressed as folds of controls. All values were mean ± SEM. * p < 0.05 ( n = 3). (B, C) Western blot analyses of CX3CR1 and LC3B (B) or IL‐1β (C) expressions in astroglial cells (C6) with treatment of CGRP at 0, 1, 2, 4, 6, and 12 h, respectively. (D, E) Western blotting analyses for CX3CR1 and LC3B (D) or IL‐1β (E) protein levels in astroglial cells (C6) with co‐treatment of CGRP (1 μM) and AA (20 μM) for 4 h. (F, G) Western blot analyses of CX3CR1 and LC3B (F) or IL‐1β (G) expression in the dorsal quadrant of L4‐L5 spinal segment ipsilateral to the operation side on 0, 1, 3, 5, 7, 10, and 14 days after CCI surgery, respectively. (H‐K) Western blot analyses of CX3CR1 or IL‐1β expression in the dorsal quadrant of L4‐L5 spinal segment ipsilateral to the operation side with CCI surgery for 5 and 7 days, respectively. Data were obtained from animals treated with daily intrathecal injection of either 1 μM CGRP (10 μl), 2 μM CGRP8‐37(10 μl), 20 μM AA (10 μl), or vehicle (10 μl) for 4 and 6 days, respectively. The mean optic densities of the proteins were calculated by normalizing to GAPDH. All values are expressed as the means ± SEMs ( n = 4).* p < 0.05 versus sham groups; # p < 0.05 versus CCI only groups

Article Snippet: The CGRP (1 μM, Tocris Bioscience), anacardic acid (AA, HAT inhibitor, 20 μM, Abcam), CGRP8‐37 (CGRP antagonist, 2 μM, MCE), or vehicle in a volume of 10 μl was injected into the spinal lumbar enlargement region through the intrathecal catheter, followed by 20 μl of saline to flush.

Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Western Blot, Injection

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: ( A ) HUMSCs, long spindle-shaped phenotype. Scale bar = 100 μm. ( B ) Trans-filter HUMSCs were stained with cresyl violet after a 6-h induction of 0, 10 −6 , 10 −7 , 10 −8 and 10 −9 mol/L CGRP. B: Images are representative of migratory fields on the underside of the membrane. Control cells were incubated in serum-free medium without CGRP. Scale bar = 100 μm. ( C ) CGRP-induced chemotactic migration of HUMSCs compared with control. Cells that migrated to the lower side of the membrane were counted under a phase-contrast light microscope (100×). Values were normalized to the control. Data represent the mean ± SEM from at least 3 independent experiments. *P < 0.05 and **P < 0.01 compared with control.

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques: Staining, Membrane, Control, Incubation, Migration, Light Microscopy

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: ( A ) Schematic model of the Dunn chamber device with the overlying coverslip, showing the position of the inner well (red), bridge (green), and outer well (blue). ( B,C ) Illustrations of FMI ( B ) and CMI ( C ), respectively. ( D,E ) Migration tracks of five representative cells in the absence of CGRP ( D ) and in the presence of 10 −6 mol/L CGRP ( E ). The starting point for each cell is the intersection between the X- and Y-axes (0, 0), and the arrow represents the direction of the outer well. ( F ) Cells induced by 10 −6 mol/L CGRP on the bridge between the inner and outer well of the chamber were observed under phase-contrast optics. The arrow points to the outer well of Dunn chamber. HUMSC migration was recorded continuously by time-lapse imaging. ( G–I ) CMI, FMI and migration speed (μm/min) for HUMSCs exposed to varying CGRP concentrations compared with control. Data are shown as the mean ± SEM from at least 3 independent experiments (*P < 0.05 and **P < 0.01).

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques: Migration, Imaging, Control

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: Cells were pretreated with 10 −6 mol/L CGRP 8–37 for 30 min, and migration was assessed with the antagonist of CGRP. ( A ) Trans-filter chemotactic migration of HUMSCs assessed using a Boyden chamber. After incubation for 6 h, cells attached to the lower surface of the filter were counted. ( B ) CMI, FMI and migration speed assessed with the Dunn chamber. Values were normalized to the control values (without CGRP stimulation). *P < 0.05 compared with control.

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques: Migration, Incubation, Control

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: ( A ) Apoptotic and dead cells were quantified through FACS analysis after staining with Annexin-V and PI. The Annexin-V+/PI− cells appeared early in the apoptotic process. Cells in death or late apoptosis were Annexin- V+/PI+. ( B ) The percentage of apoptotic and dead cells in CGRP or CGRP 8–37 groups compared with control. Cells in control group were cultured in only L-DMEM containing 10% FBS. ( C ) The CCK-8 proliferation assay showed the OD value after treatment with CGRP or CGRP 8–37 for 7 days. OD value = OD (450 nm)–OD (650 nm). Data are shown as mean ± SEM from at least three independent experiments.

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques: Staining, Control, Cell Culture, CCK-8 Assay, Proliferation Assay

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: ( A ) CGRP concentrations over time in the T9 segments of different injury models. Pre-op represents the average CGRP levels at the same segment in rats without any surgery. ( B ) Depositions of CGRP throughout the spinal cord after SCI. At 3 d post-transection surgery, the C7, T9 and L1 segments were excised for analysis. Data are presented as the mean ± SEM. *P < 0.05 versus C7.

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques:

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: ( A ) Control group; ( B ) CGRP 8–37 group; ( C ) CGRP group. Sections were double-stained for GFAP (red) and HLA (green) 7 d after transplantation. The GFAP-devoid area represents the glial scar after SCI. Engrafted HUMSCs (green with a blue nucleus) were located around the border of the scar. Left to right: rostral-caudal direction. Scale bar = 500 μm. ( D–F ) Magnification of the area marked with a rectangle in ( A–C ) respectively. Scale bar = 50 μm.

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques: Control, Staining, Transplantation Assay

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: ( A ) Cell distribution along the spinal cord after transection in the control group. Three regions were examined, C7 (rostral), T9 (injury site) and L1 (caudal). ( B ) Cells in the T9 (transection lesion site) segment of different groups (with continuous PBS, CGRP 8–37 and CGRP at the injury sites). ( C ) HUMSCs located at the lesion sites of different SCI models. HLA + and H33258 + double-stained cells in the parenchyma of the spinal cord were counted for each tissue section. *P < 0.05 and **P < 0.01 compared with the control ( A ), C7 ( B ) and contusion ( C ) groups. # P < 0.01.

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques: Control, Staining

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: HUMSCs were stimulated with various concentrations (0, 10 −6 , 10 −7 , 10 −8 and 10 −9 mol/L) of CGRP for 6 h. The active phosphorylated forms of ERK1/2, Akt, p38MAPK and SAPK/JNK (p-ERK1/2, p-Akt, p-p38MAPK, and p-SAPK/JNK) and the total proteins (ERK1/2, Akt, p38MAPK, and SAPK/JNK) were measured by densitometry using the Image Lab 4.1 software. The density ratio of phosphorylated to total ERK1/2, Akt, p38MAPK and SAPK/JNK was normalized to its related control. Equal protein loading was assessed with β-actin levels. Data are presented as the mean ± SEM. *P < 0.05 and **P < 0.01 compared with the respective CGRP-unstimulated HUMSCs.

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques: Software, Control

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: After pretreatment for 30 min, HUMSCs were added to the upper compartments of the Boyden chamber in the presence of the indicated inhibitors. ( A ) Cells that had migrated to the lower side of the filter, representative images in ( B ), were counted 6 h later. Control represents the cells only attracted to CGRP. **P < 0.01 compared with control. Scale bar = 100 μm.

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques: Control

Journal: Scientific Reports

Article Title: Calcitonin gene-related peptide is a key factor in the homing of transplanted human MSCs to sites of spinal cord injury

doi: 10.1038/srep27724

Figure Lengend Snippet: ( A ) Apoptosis and death of HUMSCs were assessed through FACS analysis. ( B ) By inhibition of Akt or p38MAPK, the percentage of apoptosis and dead cells changed. ( C ) The proliferative capacity of HUMSCs in each group was calculated using a CCK-8 assay, and the results were differently displayed on OD value columns. Data are shown as mean ± SEM from at least three independent experiments. **P < 0.01, *P < 0.05 compared with CGRP group.

Article Snippet: To verify the effects of various inhibitors on HUMSC migration, cells were pretreated with 10 −6 mol/L CGRP 8–37 (Rat, Tocris, USA), 3 × 10 −5 mol/L LY294002 (Promega, Madison, WI), 3 × 10 −5 mol/L SB203580 (Alexis Biochemical, Lausen, Switzerland), 5 × 10 −5 mol/L PD98059 (Enzo, London, UK), or 10 −5 mol/L SP600125 (Enzo) for 30 min before the migration assay.

Techniques: Inhibition, CCK-8 Assay

Journal: Science Advances

Article Title: Lung-innervating nociceptor sensory neurons promote pneumonic sepsis during carbapenem-resistant Klebsiella pneumoniae lung infection

doi: 10.1126/sciadv.adl6162

Figure Lengend Snippet: ( A ) Levels of CGRP in BALF of PBS-treated and CRKP-infected control and Trpv1 DTA mice ( n = 8 per group). ( B ) Stained sections of lungs showing β-tubulin III (TUJ1) and CGRP fibers of control and Trpv1 DTA mice. Representative images were selected from a total of nine images of three mice in each group (three images per mouse). Scale bars, 50 μm. DAPI, 4′,6-diamidino-2-phenylindole. ( C ) Left: Schematic showing organotypic culture of VG ( n = 4 VGs per well) and their stimulation with CRKP to measure CGRP release. Right: Time course of CGRP release from VG culture after stimulation with CRKP bacteria (10 6 CFU per well and 10 8 CFU per well). Unstimulated wells are VG culture with media only. Capsaicin (1.5 μM) was used as positive control. ( D ) Heatmap showing relative mRNA levels of neuropeptide receptors in CRKP-infected monocytes and neutrophils at indicated time points of infection ( n ≥ 3 per group). Un., unstimulated cells. Data in (A), (C), and (D) are the means ± SEM. Statistical analysis: Brown-Forsythe and Welch ANOVA of two-stage liner step-up procedure of Benjamini, Krieger, and Yekutieli posttests (A), one-way ANOVA with Sidak’s multiple comparison test (C), and two-way ANOVA with Sidak’s multiple comparisons posttests (D). Level of significance: * P < 0.05, ** P < 0.01, **** P < 0.0001. BioRender.com was used to create schematic in (C).

Article Snippet: Competitive CGRP inhibitor CGRP 8–37 (GenScript) (5 μg per mouse) was administered intraperitoneally into mice at 2 hours before infection.

Techniques: Infection, Control, Staining, Bacteria, Positive Control, Comparison

Journal: Science Advances

Article Title: Lung-innervating nociceptor sensory neurons promote pneumonic sepsis during carbapenem-resistant Klebsiella pneumoniae lung infection

doi: 10.1126/sciadv.adl6162

Figure Lengend Snippet: ( A ) Schematic showing monocyte and CRKP coculture in the presence and absence of neuropeptide. ( B and C ) Intracellular viable CFUs (B) and levels of ROS (C), in CRKP-infected wild-type (WT) and Ramp − / − monocytes. ( D ) Schematic of neutrophil and CRKP coculture with and without neuropeptide. ( E ) Intracellular viable CFUs in CRKP-infected WT and Ramp − / − neutrophils. ( F ) Schematic showing BMDM isolation and coculture with CRKP in the presence and absence of neuropeptides. ( G ) Intracellular viable CFUs in WT BMDMs. Data in (B), (C), (E), and (G) involve three to eight samples per group. ( H to K ) CRKP CFUs (H), proportions of myeloid subsets (I), total BAL protein (J), and panel of 13 mouse inflammatory cytokines/chemokines (K), measured in BALF of Ramp1 + / + and Ramp1 − / − mice ( n = 7 to 8 per group) at 24 hpi. ( L ) Schematic showing administration of CGRP 8–37 (5 μg per mouse) in C57BL/6 J mice and CRKP infection. ( M to P ) CFUs in whole lung (M), blood (N), liver (O), and spleen (P) of PBS-treated and CGRP 8–37 -treated mice ( n = 8 per group) after 24 hpi. Data in (B), (C), (E), (G), (H) to (K) and (M) to (P) are the means ± SEM. Statistical analysis: one-way ANOVA with Sidak’s multiple comparisons posttests [(B), (C), (E), and (G)], Mann-Whitney test [(H), (O), and (P)], unpaired t test [(J), (M), and (N)], and two-way ANOVA of two-stage liner step-up procedure of Benjamini, Krieger, and Yekutieli posttests [(I) and (K)]. Levels of significance: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. BioRender.com was used to create schematics in [(A), (D), (F), and (L)].

Article Snippet: Competitive CGRP inhibitor CGRP 8–37 (GenScript) (5 μg per mouse) was administered intraperitoneally into mice at 2 hours before infection.

Techniques: Infection, Isolation, MANN-WHITNEY

Journal: Science Advances

Article Title: Lung-innervating nociceptor sensory neurons promote pneumonic sepsis during carbapenem-resistant Klebsiella pneumoniae lung infection

doi: 10.1126/sciadv.adl6162

Figure Lengend Snippet: Lung-innervating nociceptor neurons suppress the recruitment of neutrophils and Ly6C hi monocytes to the airspaces of lungs during CRKP lung infection. CRKP infection induces vagal TRPV1 + neurons for releasing CGRP, which in turn acts on its receptor on Ly6C hi monocytes to dampen the ROS production. This leads to an increased CRKP survival in the lungs and enhanced dissemination of bacteria to vital organs. BioRender.com was used to create model.

Article Snippet: Competitive CGRP inhibitor CGRP 8–37 (GenScript) (5 μg per mouse) was administered intraperitoneally into mice at 2 hours before infection.

Techniques: Infection, Bacteria

Journal: Biomedical Reports

Article Title: Calcitonin gene-related peptide protects rats from cerebral ischemia/reperfusion injury via a mechanism of action in the MAPK pathway

doi: 10.3892/br.2016.658

Figure Lengend Snippet: Effects of CGRP on the neurobehavioral scores of rats with focal cerebral ischemia.

Article Snippet: The reagents, including CGRP, the CGRP inhibitor CGRP8–37, the ERK inhibitor PD98059, and the p38 inhibitor SB203580, were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: Biomedical Reports

Article Title: Calcitonin gene-related peptide protects rats from cerebral ischemia/reperfusion injury via a mechanism of action in the MAPK pathway

doi: 10.3892/br.2016.658

Figure Lengend Snippet: TTC staining in all the treatment groups. Normal tissues were a pink or red color, whereas the ischemic tissues were white. Compared to the sham group, the white area of TTC staining in the brain tissues in the MCAO group significantly increased. Compared to the MCAO group, the white area in the MCAO+CGRP group significantly decreased. Compared to the MCAO+CGRP group, the white area in the MCAO+CGRP+CGRP8–37, MCAO+CGRP+PD9805 and MCAO+CGRP+SB203580 groups significantly increased. TTC, 2,3,5-triphenyltetrazolium chloride; CGRP, calcitonin gene-related peptide; MCAO, middle cerebral artery occlusion.

Article Snippet: The reagents, including CGRP, the CGRP inhibitor CGRP8–37, the ERK inhibitor PD98059, and the p38 inhibitor SB203580, were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Staining

Journal: Biomedical Reports

Article Title: Calcitonin gene-related peptide protects rats from cerebral ischemia/reperfusion injury via a mechanism of action in the MAPK pathway

doi: 10.3892/br.2016.658

Figure Lengend Snippet: Effects of CGRP on the cerebral infarction area of rats with focal cerebral ischemia.

Article Snippet: The reagents, including CGRP, the CGRP inhibitor CGRP8–37, the ERK inhibitor PD98059, and the p38 inhibitor SB203580, were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: Biomedical Reports

Article Title: Calcitonin gene-related peptide protects rats from cerebral ischemia/reperfusion injury via a mechanism of action in the MAPK pathway

doi: 10.3892/br.2016.658

Figure Lengend Snippet: Western blotting results of p-JNK/JNK in all the treatment groups. Compared to the MCAO group, the p-JNK expression level significantly decreased in the MCAO+CGRP group; in addition, the JNK expression level did not significantly change. Compared to the MCAO+CGRP, the p-JNK expression levels in the MCAO+CGRP+CGRP8–37, MCAO+CGRP+PD98059 and MCAO+CGRP+SB203580 groups significantly increased, whereas no change in the JNK expression level was identified. The comparison between groups was considered to indicate significant differences, *P<0.05. p-JNK, phosphorylated c-Jun N-terminal kinase; CGRP, calcitonin gene-related peptide; MCAO, middle cerebral artery occlusion.

Article Snippet: The reagents, including CGRP, the CGRP inhibitor CGRP8–37, the ERK inhibitor PD98059, and the p38 inhibitor SB203580, were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Western Blot, Expressing

Journal: Biomedical Reports

Article Title: Calcitonin gene-related peptide protects rats from cerebral ischemia/reperfusion injury via a mechanism of action in the MAPK pathway

doi: 10.3892/br.2016.658

Figure Lengend Snippet: Western blotting results of p-p38/p38 in all the treatment groups. Compared with the MCAO group, the p-p38 expression level in the MCAO+CGRP group significantly decreased (P<0.05); in addition, the p38 expression level did not show a significant change. Compared to the MCAO+CGRP group, the p-p38 expression levels in the MCAO+CGRP+CGRP8–37, MCAO+CGRP+PD98059 and MCAO+CGRP+SB203580 groups significantly increased (P<0.05), whereas no significant change in the p38 protein expression levels was identified. The comparison between groups was considered to indicate significant differences, *P<0.05. CGRP, calcitonin gene-related peptide; MCAO, middle cerebral artery occlusion.

Article Snippet: The reagents, including CGRP, the CGRP inhibitor CGRP8–37, the ERK inhibitor PD98059, and the p38 inhibitor SB203580, were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Western Blot, Expressing

Journal: Biomedical Reports

Article Title: Calcitonin gene-related peptide protects rats from cerebral ischemia/reperfusion injury via a mechanism of action in the MAPK pathway

doi: 10.3892/br.2016.658

Figure Lengend Snippet: Western blotting results of p-ERK/ERK in all the treatment groups. Compared to the MCAO group, the p-ERK expression level significantly increased in the brain tissues in the MCAO+CGRP group; whereas no significant change in the ERK expression was identified. Compared to the MCAO+CGRP group, the expression level of the p-ERK significantly decreased in the MCAO+CGRP+CGRP8–37, MCAO+CGRP+PD98059 and MCAO+CGRP+SB203580 groups; the largest reduction was evident in the MCAO+CGRP+PD98059 group, whereas no significant change in the expression level of the ERK protein was observed. The comparison between groups was considered to indicate significant differences, *P<0.05 and **P<0.01. CGRP, calcitonin gene-related peptide; MCAO, middle cerebral artery occlusion.

Article Snippet: The reagents, including CGRP, the CGRP inhibitor CGRP8–37, the ERK inhibitor PD98059, and the p38 inhibitor SB203580, were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Western Blot, Expressing

Journal:

Article Title: CGRP receptors mediating CGRP-, adrenomedullin- and amylin-induced relaxation in porcine coronary arteries. Characterization with 'Compound 1' (WO98/11128), a non-peptide antagonist

doi: 10.1038/sj.bjp.0704210

Figure Lengend Snippet: αCGRP, βCGRP, AM and amylin concentration-reponse relationship in porcine coronary arteries. Points represent mean values and vertical lines indicate s.e.mean. Relative responses are given as percentage fraction of the initial vessel response to U46619 (10−7 M) just before they were challenged with the relaxant peptides.

Article Snippet: Human αCGRP, βCGRP, AM, amylin and the fragments αCGRP 8–37 , βCGRP 8–37 and AM 22–52 were all obtained from Bachem AG, Switzerland.

Techniques: Concentration Assay

Journal:

Article Title: CGRP receptors mediating CGRP-, adrenomedullin- and amylin-induced relaxation in porcine coronary arteries. Characterization with 'Compound 1' (WO98/11128), a non-peptide antagonist

doi: 10.1038/sj.bjp.0704210

Figure Lengend Snippet: (a–d) Vasorelaxant effect of αCGRP (10−10–10−7 M) and βCGRP (10−10–10−7 M) in the porcine LAD contracted with U46619 (10−7 M). The antagonists αCGRP8–37 (10−7–10−5 M), βCGRP8–37 (10−7–10−5 M) were added 15 min and U46619 10 min prior to the αCGRP/βCGRP challenge. Points represent mean values and vertical lines indicate s.e.mean. Relative responses are given as percentage fraction of the initial vessel response to U46619 (10−7 M) just before they were challenged with αCGRP/βCGRP.

Article Snippet: Human αCGRP, βCGRP, AM, amylin and the fragments αCGRP 8–37 , βCGRP 8–37 and AM 22–52 were all obtained from Bachem AG, Switzerland.

Techniques:

Journal:

Article Title: CGRP receptors mediating CGRP-, adrenomedullin- and amylin-induced relaxation in porcine coronary arteries. Characterization with 'Compound 1' (WO98/11128), a non-peptide antagonist

doi: 10.1038/sj.bjp.0704210

Figure Lengend Snippet: Schild plots for αCGRP8–37 and βCGRP8–37 tested with human αCGRP and βCGRP as agonists in isolated porcine LAD. The Schild plot curve for αCGRP8–37 (10−7–10−5 M) tested with αCGRP was equal to 1.48× +10.3 (r=0.62; P=0.0011). The pA2 value=7.0 (6.4–8.6). The Schild plot curve for αCGRP8–37 (10−7–10−5 M) tested with βCGRP was equal to 1.23× +7.8 (r=0.63; P=0.0007). The pA2 value=6.9 (6.2–8.7). The Schild plot curve for βCGRP8–37 (10−7–10−5 M) tested with αCGRP was equal to 1.05× +7.2 (r=0.61; P=0.0031). The pA2 value=6.3 (5.9–7.0). The Schild plot curve for βCGRP8–37 (10−6–10−5 M) tested with βCGRP was equal to 1.68× +10.0 (r=0.65; P=0.0002). The pA2 value=5.9 (5.7–6.5). Each point represents mean values and vertical lines indicate s.e.mean.

Article Snippet: Human αCGRP, βCGRP, AM, amylin and the fragments αCGRP 8–37 , βCGRP 8–37 and AM 22–52 were all obtained from Bachem AG, Switzerland.

Techniques: Isolation

Journal:

Article Title: CGRP receptors mediating CGRP-, adrenomedullin- and amylin-induced relaxation in porcine coronary arteries. Characterization with 'Compound 1' (WO98/11128), a non-peptide antagonist

doi: 10.1038/sj.bjp.0704210

Figure Lengend Snippet: (a,b) Vasorelaxant effect of αCGRP (10−10–10−7 M) and βCGRP (10−10–10−7 M) in the porcine LAD contracted with U46619 (10−7 M). The antagonist AM22–52 (10−6 M) was added 15 min and U46619 10 min prior to the αCGRP/βCGRP challenge. Points represent mean values and vertical lines indicate s.e.mean. Relative responses are given as percentage fraction of the initial vessel response to U46619 (10−7 M) just before they were challenged with αCGRP/βCGRP.

Article Snippet: Human αCGRP, βCGRP, AM, amylin and the fragments αCGRP 8–37 , βCGRP 8–37 and AM 22–52 were all obtained from Bachem AG, Switzerland.

Techniques:

Journal:

Article Title: CGRP receptors mediating CGRP-, adrenomedullin- and amylin-induced relaxation in porcine coronary arteries. Characterization with 'Compound 1' (WO98/11128), a non-peptide antagonist

doi: 10.1038/sj.bjp.0704210

Figure Lengend Snippet: Vasorelaxant effect of αCGRP in the porcine LAD contracted with U46619 (10−7 M). The non-peptide CGRP antagonist ‘Compound 1' (10−7–10−5 M) was added 15 min and U46619 10 min prior to the αCGRP (10−10–10−7 M) challenge. Points represent mean values and vertical lines indicate s.e.mean. Relative responses are given as percentage fraction of the initial vessel response to U46619 (10−7 M) just before they were challenged with αCGRP.

Article Snippet: Human αCGRP, βCGRP, AM, amylin and the fragments αCGRP 8–37 , βCGRP 8–37 and AM 22–52 were all obtained from Bachem AG, Switzerland.

Techniques: