Journal: Journal of pharmacological sciences

Article Title: Oxymatrine alleviates NSAID-associated small bowel mucosal injury by regulating MIP-1/CCR1 signalling and gut microbiota.

doi: 10.1016/j.jphs.2024.08.003

Figure Lengend Snippet: Fig. 4. OMT alleviated NSAID-associated small intestinal mucosal barrier disorder. (A) IHC analysis of the protein expression of Zo-1 and Occludin in the ileum tissues from the four groups. (B) IHC analysis of the protein expression of LYZ and CHGA in the ileum tissues from the four groups. (C) IHC analysis of the protein expression of MUC2 in the ileum tissues from the four groups.*P < 0.05 vs. the control group, #P < 0.05 vs. DS group.

Article Snippet: The primary antibodies against CHGA (ab283265, Abcam, United Kingdom), LYZ (66456-i-Ig, Proteintech, China), MUC2 (ab272692, Abcam, United Kingdom), Ki-67 (ab16667, Abcam, United Kingdom), myc (Ab32072, Abcam, United Kingdom), caspase3 (GB11009-1, Servicebio, China), CD68 (GB113109, Servicebio, China), IL-6 (GB11117, Servicebio, China), IL-1β (GB11113, Servicebio, China), TNF alpha (60291-i-Ig, Proteintech, China), Zo-1 (21773- 1-AP, Proteintech, China), Occludin (ab216327,Abcam, United Kingdom), MIP-1γ (GB112361, Servicebio, China), CCR1 (A18341, Abclonal, China), P38 MAPK (A14401, Abclonal, China), phospho-p38 MAPK (AF4001, Affinity), phospho–NF–KB p65 (AP0944, Abclonal), NF-KB p65 (8242,CST), MIP-1α (bs-1045r, BIOSS), CCR1 (bs-1169r, BIOSS) and GAPDH (GB15004, Servicebio) were used in the follow-up study.

Techniques: Expressing, Control

Journal: Scientific Reports

Article Title: fMRI indicates cortical activation through TRPV1 modulation during acute gouty attacks

doi: 10.1038/s41598-019-48656-6

Figure Lengend Snippet: Cortical TRPV1 expression was enhanced by gouty arthritis. ( A ) ROI selection. ( B ) Cortical TRPV1 expression on the MSU-affected and control sides. ( C ) TRPV1 immunoreactivity intensity. ( D ) TRPV1 immunoreactivity coverage. ( E ) Immunoprecipitation of beta-tubulin with TRPV1 immunoblotting, which demonstrates a strong interaction between TRPV1 and neuronal filament proteins. The cropped images of the gels are used in the figure, and images of the full-length gels are presented in Supplementary Fig. S2. (***Indicates statistically significant difference with p < 0.001 after ANOVA). Scale bar = 50 µm.

Article Snippet: The primary antibodies used were: anti-TRPV1 (1:2000; PC420; Merck Millipore, Darmstadt, Germany); anti-COX-2 (1:1000; Thermo Fisher Scientific); β-tubulin (1:2000; MCA2703; Bio-Rad Laboratories); β-actin (1:4000; sc-47778; Santa Cruz Biotechnology).

Techniques: Expressing, Selection, Control, Immunoprecipitation, Western Blot

Journal: Neurochemistry international

Article Title: Red nucleus mGluR2 but not mGluR3 mediates inhibitory effect in the development of SNI-induced neuropathological pain by suppressing the expressions of TNF-α and IL-1β.

doi: 10.1016/j.neuint.2024.105840

Figure Lengend Snippet: Fig. 1. Reduced protein expression of mGluR2 but not mGluR3 in the RN of SNI rats. A: Neuropathological pain induced by SNI (***P < 0.001, compared with sham- operated rats). B–C: Western blotting indicated that mGluR2 and mGluR3 were constitutively expressed in the RN of normal rats, mGluR2 rather than mGluR3 was reduced at 2 weeks post-SNI (n = 6 rats per group; ***P < 0.001). D–E: Immunohistochemistry indicated that mGluR2 and mGluR3 were constitutively expressed in the RN of normal rats, mGluR2 rather than mGluR3 was reduced at 2 weeks post-SNI (n = 4 rats per group; **P < 0.01). Scale bars = 50 μm.

Article Snippet: Selective mGluR2/3 agonist LY379268 (4.0 μg/μl, Glpbio, USA) and mGluR3 antagonist β-NAAG (30 μg/μl, MedChemExpress, USA) were dissolved in physiological saline, mGluR2/3 antagonist EGLU (1.8 μg/μl, MedChemExpress) was dissolved in a solution of 3% NaOH (1 M) and 97% physiological saline.

Techniques: Expressing, Western Blot, Immunohistochemistry

Journal: Neurochemistry international

Article Title: Red nucleus mGluR2 but not mGluR3 mediates inhibitory effect in the development of SNI-induced neuropathological pain by suppressing the expressions of TNF-α and IL-1β.

doi: 10.1016/j.neuint.2024.105840

Figure Lengend Snippet: Fig. 2. Activation of red nucleus mGluR2 but not mGluR3 inhibits the development of SNI-induced neuropathological pain. A: Intrarubral injection of mGluR2/3 agonist LY379268 (2.0 μg) at 2 weeks post-SNI significantly attenuated neuropathological pain as compared with vehicle alone. B: Intrarubral injection of mGluR2/3 antagonist EGLU (0.9 μg) at 10 min after LY379268 delivery significantly reversed the analgesic effect of LY379268, and did not show significant difference as compared with vehicle alone. C: Intrarubral injection of mGluR3 antagonist β-NAAG (15 μg) at 10 min after LY379268 delivery did not affect the analgesic effect of LY379268, and displayed significant difference as compared with vehicle alone. D–F: Footprint test showed that intrarubral injection of LY379268, LY379268 plus EGLU or LY379268 plus β-NAAG had no impact on the movement of rats. *P < 0.05, **P < 0.01 and ***P < 0.001, compared with vehicle.

Article Snippet: Selective mGluR2/3 agonist LY379268 (4.0 μg/μl, Glpbio, USA) and mGluR3 antagonist β-NAAG (30 μg/μl, MedChemExpress, USA) were dissolved in physiological saline, mGluR2/3 antagonist EGLU (1.8 μg/μl, MedChemExpress) was dissolved in a solution of 3% NaOH (1 M) and 97% physiological saline.

Techniques: Activation Assay, Injection

Journal: Neurochemistry international

Article Title: Red nucleus mGluR2 but not mGluR3 mediates inhibitory effect in the development of SNI-induced neuropathological pain by suppressing the expressions of TNF-α and IL-1β.

doi: 10.1016/j.neuint.2024.105840

Figure Lengend Snippet: Fig. 3. Blockade of red nucleus mGluR2 but not mGluR3 in normal rats induces mechanical allodynia. A: Intrarubral injection of mGluR2/3 agonist LY379268 (2.0 μg) did not change the PWT of normal rats. B: Intrarubral injection of mGluR2/3 antagonist EGLU (0.9 μg) induced a significant mechanical allodynia in normal rats as compared with vehicle alone. C: Intrarubral injection of mGluR3 antagonist β-NAAG (15 μg) did not influence the PWT of normal rats. D–F: Footprint test demonstrated that intrarubral injection of LY379268, EGLU or β-NAAG did not influence the movement of rats. *P < 0.05 and ***P < 0.001, compared with vehicle.

Article Snippet: Selective mGluR2/3 agonist LY379268 (4.0 μg/μl, Glpbio, USA) and mGluR3 antagonist β-NAAG (30 μg/μl, MedChemExpress, USA) were dissolved in physiological saline, mGluR2/3 antagonist EGLU (1.8 μg/μl, MedChemExpress) was dissolved in a solution of 3% NaOH (1 M) and 97% physiological saline.

Techniques: Injection

Journal: Neurochemistry international

Article Title: Red nucleus mGluR2 but not mGluR3 mediates inhibitory effect in the development of SNI-induced neuropathological pain by suppressing the expressions of TNF-α and IL-1β.

doi: 10.1016/j.neuint.2024.105840

Figure Lengend Snippet: Fig. 4. Red nucleus mGluR2 but not mGluR3 mediates inhibitory effect in the development of SNI-induced neuropathological pain by suppressing the expression of TNF-α. A–B: Western blotting indicated that red nucleus TNF-α was elevated at 2 weeks post-SNI, intrarubral injection of mGluR2/3 agonist LY379268 (2.0 μg) at 2 weeks post-SNI inhibited the overexpression of TNF-α, this effect was reversed by mGluR2/3 antagonist EGLU (0.9 μg) instead of selective mGluR3 antagonist β-NAAG (15 μg) (n = 6 rats per group). C–D: Western blotting displayed that intrarubral injection of LY379268 (2.0 μg) did not influence the protein expression of TNF-α in normal rats, while intrarubral injection of EGLU (0.9 μg) rather than β-NAAG (15 μg) significantly enhanced the expression of TNF-α (n = 6 rats per group). E–G: Immunohistochemistry showed that red nucleus TNF-α was elevated at 2 weeks post-SNI, intrarubral injection of LY379268 at 2 weeks post-SNI inhibited the overexpression of TNF-α, this effect was reversed by EGLU instead of β-NAAG. Intrarubral injection of LY379268 did not influence the protein expression of TNF-α in normal rats, while intrarubral injection of EGLU rather than β-NAAG significantly enhanced the expression of TNF-α (n = 4 rats per group). *P < 0.05, **P < 0.01 and ***P < 0.001. Scale bars = 50 μm.

Article Snippet: Selective mGluR2/3 agonist LY379268 (4.0 μg/μl, Glpbio, USA) and mGluR3 antagonist β-NAAG (30 μg/μl, MedChemExpress, USA) were dissolved in physiological saline, mGluR2/3 antagonist EGLU (1.8 μg/μl, MedChemExpress) was dissolved in a solution of 3% NaOH (1 M) and 97% physiological saline.

Techniques: Expressing, Western Blot, Injection, Over Expression, Immunohistochemistry

Journal: Neurochemistry international

Article Title: Red nucleus mGluR2 but not mGluR3 mediates inhibitory effect in the development of SNI-induced neuropathological pain by suppressing the expressions of TNF-α and IL-1β.

doi: 10.1016/j.neuint.2024.105840

Figure Lengend Snippet: Fig. 5. Red nucleus mGluR2 but not mGluR3 mediates inhibitory effect in the development of SNI-induced neuropathological pain by suppressing the expression of IL-1β. A–B: Western blotting showed that red nucleus IL-1β was elevated at 2 weeks post-SNI, intrarubral injection of mGluR2/3 agonist LY379268 (2.0 μg) at 2 weeks post-SNI inhibited the overexpression of IL-1β, this effect was reversed by mGluR2/3 antagonist EGLU (0.9 μg) instead of selective mGluR3 antagonist β-NAAG (15 μg) (n = 6 rats per group). C–D: Western blotting displayed that intrarubral injection of LY379268 (2.0 μg) did not influence the protein expression of IL-1β in normal rats, while intrarubral injection of EGLU (0.9 μg) rather than β-NAAG (15 μg) significantly enhanced the expression of IL-1β (n = 6 rats per group). E–G: Immu nohistochemistry showed that red nucleus IL-1β was elevated at 2 weeks post-SNI, intrarubral injection of LY379268 at 2 weeks post-SNI inhibited the overexpression of IL-1β, this effect was reversed by EGLU instead of β-NAAG. Intrarubral injection of LY379268 did not influence the protein expression of IL-1β in normal rats, while intrarubral injection of EGLU rather than β-NAAG significantly enhanced the expression of IL-1β (n = 4 rats per group). **P < 0.01 and ***P < 0.001. Scale bars = 50 μm.

Article Snippet: Selective mGluR2/3 agonist LY379268 (4.0 μg/μl, Glpbio, USA) and mGluR3 antagonist β-NAAG (30 μg/μl, MedChemExpress, USA) were dissolved in physiological saline, mGluR2/3 antagonist EGLU (1.8 μg/μl, MedChemExpress) was dissolved in a solution of 3% NaOH (1 M) and 97% physiological saline.

Techniques: Expressing, Western Blot, Injection, Over Expression

Journal: Neurochemistry international

Article Title: Red nucleus mGluR2 but not mGluR3 mediates inhibitory effect in the development of SNI-induced neuropathological pain by suppressing the expressions of TNF-α and IL-1β.

doi: 10.1016/j.neuint.2024.105840

Figure Lengend Snippet: Fig. 6. Schematics presents the underlying mechanisms of red nucleus mGluR II in pain modulation. A: mGluR II(mGluR2 and mGluR3) were constitutively expressed in the RN of normal rats. Blockade of red nucleus mGluR2 rather than mGluR3 in normal rats induced mechanical allodynia. Red nucleus mGluR2 but not mGluR3 mediated antinociceptive effect in normal rats through negatively modulating the expressions of pro-inflammatory factors TNF-α and IL-1β. B: Red nucleus mGluR2 but not mGluR3 was reduced in SNI rats, while TNF-α and IL-1β were elevated in SNI rats. Activation of mGluR2 rather than mGluR3 alleviated SNI-induced neuropathological pain through inhibiting the expressions of TNF-α and IL-1β. Glu, glutamate.

Article Snippet: Selective mGluR2/3 agonist LY379268 (4.0 μg/μl, Glpbio, USA) and mGluR3 antagonist β-NAAG (30 μg/μl, MedChemExpress, USA) were dissolved in physiological saline, mGluR2/3 antagonist EGLU (1.8 μg/μl, MedChemExpress) was dissolved in a solution of 3% NaOH (1 M) and 97% physiological saline.

Techniques: Activation Assay

Journal: Cell

Article Title: BMAL1-Driven Tissue Clocks Respond Independently to Light to Maintain Homeostasis.

doi: 10.1016/j.cell.2019.05.009

Figure Lengend Snippet: Figure 5. Epidermal BMAL1 Is Required to Prevent Increased Differentiation (A) Kaplan Meier survival curve for WT, RE, and KO mice (no significant difference between KO and RE, p = 0.2730). (B) Weight curve for WT, RE, and KO mice; data are represented as mean ± SD. (C) Epidermal cornification measured as thickness of cornified layer; p(RE versus WT) = 1.97 3 103, p(RE versus KO) = 1.33 3 106, p(WT versus KO) = 8.79 3 1010); data are represented as mean ± SD; scale bar, 100 mm.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-BMAL1 Abcam Cat# ab93806; RRID:AB_10675117 Rabbit anti-phosphorylated BMAL1 (Ser42) Cell Signaling Cat# 13936 Rabbit polyclonal anti-acetylated BMAL1 (Lys538) Merck Millipore Cat# AB15396; RRID:AB_1977054 Mouse monoclonal anti-ACTIN Sigma-Aldrich Cat# A5228; RRID:AB_262054 Mouse monoclonal anti-TUBULIN Sigma-Aldrich Cat# T7816; RRID:AB_261770 Rabbit polyclonal anti-RFP Rockland Cat# 600-401-379; RRID:AB_2209751 AffiniPure Goat anti-Rabbit IgG Jackson ImmunoResearch Labs Cat# 111-005-003; RRID:AB_2337913 BrightVision Poly-HRP-anti Goat IgG Biotin-free ImmunoLogic Cat# DPVG 55HRP Rabbit polyclonal anti-LORICRIN Abcam Cat# ab85679; RRID:AB_2134912 Chicken polyclonal anti-KERATIN 14 BioLegend Cat# 906001; RRID:AB_2565055 Goat anti-Chicken IgY, Alexa Fluor 647 Thermo Fisher Scientific Cat# A-21449; RRID:AB_2535866 Goat anti-Rabbit IgG, Alexa Fluor 568 Thermo Fisher Scientific Cat# A-11036; RRID:AB_10563566 Chemicals, Peptides, and Recombinant Proteins SsoAdvanced Universal SYBR Green Supermix BioRad Cat# 1725270 Taqman Gene Expression Master Mix Thermo Fisher Cat# 4369016 Critical Commercial Assays Applied Biosystems High Capacity cDNA Reverse Transcription Kit Thermo Fisher Cat# 4368814 iScript cDNA Synthesis Kit BioRad Cat# 1708890 Deposited Data L/D RNA-sequencing dataset Gene Expression Omnibus GEO: GSE115104 D/D RNA-sequencing dataset Gene Expression Omnibus GEO: GSE114943 Experimental Models: Organisms/Strains Mouse:Wild Type –Bmal1wt/wt;K14-cretg/wt this paper N/A Mouse: Bmal1 knockout – Bmal1stopFL/stopFL; K14-crewt/wt this paper N/A Mouse: Bmal1 Epidermis-Reconstituted – Bmal1stopFL/stopFL; K14-cretg/wt this paper N/A Mouse:Wild Type –Bmal1wt/wt;Alfp-cretg/wt this paper N/A Mouse: Bmal1 knockout – Bmal1stopFL/stopFL; Alfp-crewt/wt this paper N/A Mouse: Bmal1 Hepatocyte-Reconstituted (Liver-RE) – Bmal1stopFL/stopFL; Alfp-cretg/wt this paper N/A Oligonucleotides qPCR Primers This paper Table S3 Software and Algorithms Jonckheere-Terpstra-Kendall (JTK_CYCLE) algorithm Hughes et al., 2010 N/A Trimmomatic (version 0.36) Bolger et al., 2014 N/A TopHat (version 2.1.1) Kim et al., 2013a https://ccb.jhu.edu/software/ tophat/index.shtml (Continued on next page) e1 Cell 177, 1436–1447.e1–e5, May 30, 2019

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