Journal: Molecular Pain

Article Title: Neuropeptide deficient mice have attenuated nociceptive, vascular, and inflammatory changes in a tibia fracture model of complex regional pain syndrome

doi: 10.1186/1744-8069-8-85

Figure Lengend Snippet: Determination of hindpaw skin cytokine and NGF levels at 3 weeks after fracture (FX) in WT, SP deficient (SP KO, Tac1 −/− ), and CGRP receptor deficient (RAMP1 KO, RAMP1 −/− ) mice. TNFα (A, E), IL-1β (B, F), IL-6 (C, G) and NGF (D, H) production in hindpaw skin were determined by Bio-Rad bead suspension array (for cytokines) and EIA (for NGF). Baseline cytokine and NGF levels were the same in all 3 groups of mice. Tibia fracture induced a significant increase in all three cytokine levels and NGF production in WT FX group vs WT Controls, however, fracture did not induce increased TNFα ( A ), IL-1β ( B ), or NGF ( D ) levels in SP KO FX mice compared to unfractured SP KO Control mice. The IL-6 ( C ) levels were elevated in the SP KO FX mice, compared to unfractured SP KO Control mice, but the IL-6 increase the SP KO FX mice was considerable reduced compared to the IL-6 levels in the WT FX mice. A similar pattern was observed in the RAMP1 KO mice for TNFα ( E ), IL-1β ( F ), or NGF ( H ), except the increase in IL-6 ( G ) levels in the RAMP1 KO FX mice was even greater than the increase observed in the WT FX mice. Data are expressed as mean values (pg/mg protein) ± SE (n = 8 per group). *P < 0.05, **P < 0.001, and ***P < 0.0001 vs the respective unfractured Control mice; #P < 0.05, ##P < 0.001, and ###P < 0.0001 vs WT FX mice.

Article Snippet: For IL-6 immunostaining rabbit anti human IL-6 antibody (1:400, LifeSpan Biosciences Inc) was labeled with donkey anti-rabbit IgG (1:500) conjugated with Dylight 488 secondary antibodies (Jackson ImmunoResearch).

Techniques:

Journal: Molecular Pain

Article Title: Neuropeptide deficient mice have attenuated nociceptive, vascular, and inflammatory changes in a tibia fracture model of complex regional pain syndrome

doi: 10.1186/1744-8069-8-85

Figure Lengend Snippet: Fluorescence photomicrographs of keratin (red), IL-1β (green), and IL-6 (green) immunostaining in the plantar hindpaw skin at 3 weeks post-fracture (FX). The top two rows of panels show keratinocyte and IL-1β immunostaining from a wildtype (WT) unfractured control (Cont) mouse (first row) and the second row panels are from a WT fracture mouse. The bottom two rows of panels show keratinocyte and IL-6 immunostaining from a WT unfractured control mouse (third row) and the bottom row panels are from a WT fracture mouse. Double labeling demonstrates fracture-induced IL-1β and IL-6 protein up-regulation localized within the epidermal keratinocyte layer of the WT mice. Scale bar = 20 μm.

Article Snippet: For IL-6 immunostaining rabbit anti human IL-6 antibody (1:400, LifeSpan Biosciences Inc) was labeled with donkey anti-rabbit IgG (1:500) conjugated with Dylight 488 secondary antibodies (Jackson ImmunoResearch).

Techniques: Fluorescence, Immunostaining, Labeling

Journal: Molecular Pain

Article Title: Neuropeptide deficient mice have attenuated nociceptive, vascular, and inflammatory changes in a tibia fracture model of complex regional pain syndrome

doi: 10.1186/1744-8069-8-85

Figure Lengend Snippet: After baseline testing, WT mice underwent a right distal tibia fracture (FX) and the hindlimb was casted for 3 weeks, then the cast was removed and the next day the animals retested (n = 12 per cohort). Then the FX mice were either subcutaneously injected with the IL-6 receptor antagonist TB-2-081 (FX + TB-2-081, n = 8) or vehicle (FX + Vehicle, n = 6). A control cohort of WT mice did not undergo tibia fracture (Control, n = 12). At 15 minutes post-injection the mice were retested. The IL-6 receptor antagonist partially inhibited the development of von Frey allodynia ( A ) and hindlimb unweighting ( B ) in the fracture mice, when compared to the extent of allodynia and unweighting observed in the vehicle injected mice after fracture. The IL-6 receptor antagonist failed to reverse the hindpaw warmth ( C ) and edema ( D ) that was observed in the vehicle injected fracture mice. ***P < 0.001 vs Control, ##P < 0.01, and ###P < 0.001 vs FX + Vehicle.

Article Snippet: For IL-6 immunostaining rabbit anti human IL-6 antibody (1:400, LifeSpan Biosciences Inc) was labeled with donkey anti-rabbit IgG (1:500) conjugated with Dylight 488 secondary antibodies (Jackson ImmunoResearch).

Techniques: Injection

Journal: Frontiers in Pharmacology

Article Title: Effects of Inonotus obliquus on ameliorating podocyte injury in ORG mice through TNF pathway and prediction of active compounds

doi: 10.3389/fphar.2024.1426917

Figure Lengend Snippet: The expressions of renal TNF-α and IL-6 (×200). CON: the control group; ORG: the obesity-related glomerulopathy model; IO_L: the model group treated with Inonotus obliquus (75 mg/kg/day); IO_H: the model group treated with I . obliquus (150 mg/kg/day). Data were presented as mean ± SD. # p < 0.05 vs. the CON; * p < 0.05 vs. the ORG.

Article Snippet: After deparaffinization, antigen retrieval and blocking with goat serum, the section was incubating with anti-IL-6 or anti-TNF-α rabbit polyclonal antibodies (Servicebio, Wuhan, China) overnight at 4°C.

Techniques: Control

Journal: Frontiers in Pharmacology

Article Title: Effects of Inonotus obliquus on ameliorating podocyte injury in ORG mice through TNF pathway and prediction of active compounds

doi: 10.3389/fphar.2024.1426917

Figure Lengend Snippet: Molecular docking of IO against ORG based on active compounds and targets. (A) The overlapping compounds between IO against ORG and IO absorbed in blood. (B) Classification of active compounds of IO against ORG. (C) Structural formula of interpene F and trametenolic acid. (D) Molecular docking of interpene F and trametenolic acid with TNF-α and IL-6. IO: Inonotus obliquus ; ORG: obesity-related glomerulopathy.

Article Snippet: After deparaffinization, antigen retrieval and blocking with goat serum, the section was incubating with anti-IL-6 or anti-TNF-α rabbit polyclonal antibodies (Servicebio, Wuhan, China) overnight at 4°C.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Effects of Inonotus obliquus on ameliorating podocyte injury in ORG mice through TNF pathway and prediction of active compounds

doi: 10.3389/fphar.2024.1426917

Figure Lengend Snippet: Downstream regulatory mechanisms of TNF-α and IL-6. TNFR1: TNF-α receptor 1; TRAF1: TNF receptor associated factor 1; TRADD: TNFR1-associated death domain protein; MAPK: mitogen-activated protein kinase; IL-6R: Interleukin 6 receptor; JAK: Janus-activated kinase; STAT3: signal transducer and activator of transcription 3.

Article Snippet: After deparaffinization, antigen retrieval and blocking with goat serum, the section was incubating with anti-IL-6 or anti-TNF-α rabbit polyclonal antibodies (Servicebio, Wuhan, China) overnight at 4°C.

Techniques: