human il 6 5 20 100 ng ml 16 h  (R&D Systems)

Journal: Scientific Reports

Article Title: Anti-interleukin-6 receptor antibody treatment ameliorates postoperative adhesion formation

doi: 10.1038/s41598-019-54175-1

Figure Lengend Snippet: Proinflammatory responses prior to pro-fibrotic alterations. ( a,c ) Cecum lesions were sampled from each experimental group (3–5 mice/group) at the indicated time points post operation, followed by qRT-PCR assessment of expression of proinflammatory cytokine and pro-fibrotic molecule genes ( a ) and by immunostaining of phosphorylated transcription factors with ImageJ analysis data ( c,d ) Ifng , interferon γ; Il6 , interleukin-6; Tnf , tumor necrosis factor-α; Pai1 , plasminogen activator inhibitor type 1; Col1a1 , collagen 1α1; Tgfb1 , tumor growth factor- β1. ( b ) Peritoneal fluid and serum were sampled from human patients at the indicated time points following the beginning of laparotomy, and IL-6 levels were measured using ELISA. Representative photos are shown. Data at 0 day or hour postoperation indicated those in untreated control mice. Data are shown as mean ± SD. * p < 0.05 (Dunnett’s test). Daggers indicate serosa; open brackets indicate adhesion lesions. All experiments were independently repeated twice.

Article Snippet: Recombinant (r) human (h) TNF-α (21-TA), rhIL-6 (206-IL), and rhTGF-β1 (240-B) were purchased from R& D Systems. rhsIL-6Rα(00–06RC) was obtained from Peprotech.

Techniques: Quantitative RT-PCR, Expressing, Immunostaining, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Anti-interleukin-6 receptor antibody treatment ameliorates postoperative adhesion formation

doi: 10.1038/s41598-019-54175-1

Figure Lengend Snippet: Production of pro-fibrotic molecules by human neutrophils in response to proinflammatory cytokines. Expression of IL6 , TNF , and TGFB1 was determined in human neutrophils stimulated with TNF-α, IL-6, or TGF-β1 using qRT-PCR ( a ). Human mesothelial cells (MeT5A cells) were incubated with TNF-α, IL-6 plus soluble IL-6Rα (sIL-6Rα), TGF-β1, or IFN-γ, followed by measurement of IL6 ( b ), TNF ( b ), CXCL2 ( b ), TGFB1 ( b ), or COL1A1 expression by qRT-PCR ( c ). Three independent experiments were performed. Data are shown as mean ± SEM. * p < 0.05 (Dunnett’s test).

Article Snippet: Recombinant (r) human (h) TNF-α (21-TA), rhIL-6 (206-IL), and rhTGF-β1 (240-B) were purchased from R& D Systems. rhsIL-6Rα(00–06RC) was obtained from Peprotech.

Techniques: Expressing, Quantitative RT-PCR, Incubation

Journal: Scientific Reports

Article Title: Anti-interleukin-6 receptor antibody treatment ameliorates postoperative adhesion formation

doi: 10.1038/s41598-019-54175-1

Figure Lengend Snippet: Proposed model for the role of IL-6 in the induction of postoperative adhesion formation. Upon surgery-induced injury, serosa-resident natural killer T (NKT) cells produce IFN-γ, thereby inducing PAI-1, which in turn facilitates fibrin deposition by preventing fibrin degradation as shown previously . Subsequently, mesothelial cells produce CXCL2 and IL-6, which recruit and activate neutrophils, respectively. The neutrophils produce TNF-α in response to IL-6. Both mesothelial cells and neutrophils produce TNF-α in response to TNF-α. Thus, IL-6 signaling plays a central role in activating the TNF-α-amplifying positive-feedback circuits. The final steps of the initiation of adhesion formation occur when TNF-α induces neutrophils, but not mesothelial cells, to produce TGF-β1. TGF-β1 subsequently activates mesothelial cells to transdifferentiate into myofibroblasts that produce fibrosis-promoting molecules such as collagen.

Article Snippet: Recombinant (r) human (h) TNF-α (21-TA), rhIL-6 (206-IL), and rhTGF-β1 (240-B) were purchased from R& D Systems. rhsIL-6Rα(00–06RC) was obtained from Peprotech.

Techniques:

Journal: Journal of Inflammation (London, England)

Article Title: TNF-alpha and IL-6 inhibit apolipoprotein A-IV production induced by linoleic acid in human intestinal Caco2 cells

doi: 10.1186/s12950-015-0069-0

Figure Lengend Snippet: Effects of cytokines on apoA-IV and apoC-III gene expression. (A) Effects of LA and cytokines on apoA-IV gene expression. Differentiated Caco2 cells were pre-treated with or without r-h-TNF-α (20 ng/ml) or r-h-IL-6 (20 ng/ml) or the combination of these two cytokines for 72 h before changing into medium with or without 1 mM LA for 24 h. (B) Effects of LA and cytokines on apoA-IV protein secretion. Differentiated Caco2 cells were pre-treated with or without r-h-TNF-α (20 ng/ml) or r-h-IL-6 (20 ng/ml) or the combination of these two cytokines for 72 h before changing into medium with or without 0.25 or 0.5 mM LA for 24 h. The apoA-IV proteins in culture medium secreted from the cells were measured by ELISA. (C) Effects of LA and cytokines on apoC-III expression. The samples were processed at the same ways described at (A). The mRNA levels of both apoA-IV and apoC-III were measured by real time RT-PCR. * P < 0.05, ** P < 0.01, *** P < 0.001 vs . vehicle control; and ## P < 0.01 ### P < 0.001 vs. LA.

Article Snippet: Recombinant human IL-6 (r-h-IL-6) and recombinant human TNF-α (r-h-TNF-α) were purchased from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Journal of Inflammation (London, England)

Article Title: TNF-alpha and IL-6 inhibit apolipoprotein A-IV production induced by linoleic acid in human intestinal Caco2 cells

doi: 10.1186/s12950-015-0069-0

Figure Lengend Snippet: Effect of cytokines on gene expression of Il6 (A) and Tnfα (B) in Caco2 cells. Differentiated Caco2 cells were pre-treated with or without r-h-TNF-α (20 ng/ml), r-h-IL-6 (20 ng/ml) or their combination for 72 h before changing into the medium with or without LA (1 mM) for 24 h. The mRNA levels of both Il6 and Tnfα were measured by real time RT-PCR. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. vehicle control; and # P <0.05, ### P <0.001 vs . LA.

Article Snippet: Recombinant human IL-6 (r-h-IL-6) and recombinant human TNF-α (r-h-TNF-α) were purchased from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Expressing, Quantitative RT-PCR