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Interleukin(IL)-36α and IL-36γ Induce Proinflammatory Mediators from Human Colonic Subepithelial Myofibroblasts.

Kanda T, Nishida A, Takahashi K, Hidaka K, Imaeda H, Inatomi O, Bamba S, Sugimoto M, Andoh A - Front Med (Lausanne) (2015)

Bottom Line: The mRNA expression of proinflammatory mediators induced by IL-36α and/or IL-36γ was significantly suppressed by transfection of siRNA for MyD88 or IRAK1.Both inhibitors of mitogen activated protein kinases and siRNAs specific for NF-κBp65 significantly reduced the expression of IL-6 and CXC chemokines induced by IL-36α and/or IL-36γ.These results suggest that IL-36α and IL-36γ contribute to gut inflammation through the induction of proinflammatory mediators.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Shiga University of Medical Science , Otsu , Japan.

ABSTRACT

Background: Interleukin (IL)-36 cytokines are recently reported member of the IL-1 cytokine family. However, there is little information regarding the association between IL-36 cytokines and gut inflammation. In the present study, we investigated the biological activity of IL-36α and IL-36γ using human colonic subepithelial myofibroblasts (SEMFs).

Methods: The mRNA expression and the protein expression of target molecules in SEMFs were evaluated using real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The intracellular signaling of IL-36 cytokines was analyzed using Western blot analysis and small interfering RNAs (siRNAs) specific for MyD88 adaptor proteins (MyD88 and IRAK1) and NF-κB p65.

Results: IL-36α and IL-36γ significantly enhanced the secretion of IL-6 and CXC chemokines (CXCL1, CXCL2, and CXCL8) by SEMFs. The combination of IL-36α/γ and IL-17A or of IL-36α/γ and tumor necrosis factor-α showed a synergistic effect on the induction of IL-6 and CXC chemokines. The mRNA expression of proinflammatory mediators induced by IL-36α and/or IL-36γ was significantly suppressed by transfection of siRNA for MyD88 or IRAK1. Both inhibitors of mitogen activated protein kinases and siRNAs specific for NF-κBp65 significantly reduced the expression of IL-6 and CXC chemokines induced by IL-36α and/or IL-36γ.

Conclusion: These results suggest that IL-36α and IL-36γ contribute to gut inflammation through the induction of proinflammatory mediators.

No MeSH data available.


Related in: MedlinePlus

Effects of combination of IL-36α/γ plus IL-17A or that of IL-36α/γ plus TNF-α on the induction of IL-6 and CXC chemokines. SEMFs were stimulated with combination of IL-36α/γ plus IL-17A or that of IL-36α/γ plus TNF-α for 24 h [(A) IL-36α plus IL-17A or IL-36α plus TNF-α, (B) IL-36γ plus IL-17A or IL-36γ plus TNF-α]. The mRNA expression of IL-6 and CXC chemokines (CXCL1, CXCL2, and CXCL8) was then determined using real-time PCR. The mRNA expression for IL-6 and chemokine was converted to a value relative to β-actin mRNA expression and presented as fold-increase relative to the results for medium alone (no stimulation). Data are expressed as means ± SD of four independent experiments. *P < 0.05, **P < 0.01; significant differences from the values for IL-17A or TNF-α stimulation.
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Figure 3: Effects of combination of IL-36α/γ plus IL-17A or that of IL-36α/γ plus TNF-α on the induction of IL-6 and CXC chemokines. SEMFs were stimulated with combination of IL-36α/γ plus IL-17A or that of IL-36α/γ plus TNF-α for 24 h [(A) IL-36α plus IL-17A or IL-36α plus TNF-α, (B) IL-36γ plus IL-17A or IL-36γ plus TNF-α]. The mRNA expression of IL-6 and CXC chemokines (CXCL1, CXCL2, and CXCL8) was then determined using real-time PCR. The mRNA expression for IL-6 and chemokine was converted to a value relative to β-actin mRNA expression and presented as fold-increase relative to the results for medium alone (no stimulation). Data are expressed as means ± SD of four independent experiments. *P < 0.05, **P < 0.01; significant differences from the values for IL-17A or TNF-α stimulation.

Mentions: The combined effect of IL-36 and IL-17A or IL-36 and TNFα on the mRNA production of inflammatory mediators was evaluated using real-time PCR. As shown in Figure 3A, the combination of IL-36α and IL-17A or IL-36α and TNFα synergistically enhanced the mRNA expression of IL-6 and CXC chemokines. Similarly, simultaneous stimulation with IL-36γ and IL-17A or IL-36γ and TNFα synergistically upregulated the mRNA expression of IL-6 and CXC chemokines (Figure 3B).


Interleukin(IL)-36α and IL-36γ Induce Proinflammatory Mediators from Human Colonic Subepithelial Myofibroblasts.

Kanda T, Nishida A, Takahashi K, Hidaka K, Imaeda H, Inatomi O, Bamba S, Sugimoto M, Andoh A - Front Med (Lausanne) (2015)

Effects of combination of IL-36α/γ plus IL-17A or that of IL-36α/γ plus TNF-α on the induction of IL-6 and CXC chemokines. SEMFs were stimulated with combination of IL-36α/γ plus IL-17A or that of IL-36α/γ plus TNF-α for 24 h [(A) IL-36α plus IL-17A or IL-36α plus TNF-α, (B) IL-36γ plus IL-17A or IL-36γ plus TNF-α]. The mRNA expression of IL-6 and CXC chemokines (CXCL1, CXCL2, and CXCL8) was then determined using real-time PCR. The mRNA expression for IL-6 and chemokine was converted to a value relative to β-actin mRNA expression and presented as fold-increase relative to the results for medium alone (no stimulation). Data are expressed as means ± SD of four independent experiments. *P < 0.05, **P < 0.01; significant differences from the values for IL-17A or TNF-α stimulation.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4585048&req=5

Figure 3: Effects of combination of IL-36α/γ plus IL-17A or that of IL-36α/γ plus TNF-α on the induction of IL-6 and CXC chemokines. SEMFs were stimulated with combination of IL-36α/γ plus IL-17A or that of IL-36α/γ plus TNF-α for 24 h [(A) IL-36α plus IL-17A or IL-36α plus TNF-α, (B) IL-36γ plus IL-17A or IL-36γ plus TNF-α]. The mRNA expression of IL-6 and CXC chemokines (CXCL1, CXCL2, and CXCL8) was then determined using real-time PCR. The mRNA expression for IL-6 and chemokine was converted to a value relative to β-actin mRNA expression and presented as fold-increase relative to the results for medium alone (no stimulation). Data are expressed as means ± SD of four independent experiments. *P < 0.05, **P < 0.01; significant differences from the values for IL-17A or TNF-α stimulation.
Mentions: The combined effect of IL-36 and IL-17A or IL-36 and TNFα on the mRNA production of inflammatory mediators was evaluated using real-time PCR. As shown in Figure 3A, the combination of IL-36α and IL-17A or IL-36α and TNFα synergistically enhanced the mRNA expression of IL-6 and CXC chemokines. Similarly, simultaneous stimulation with IL-36γ and IL-17A or IL-36γ and TNFα synergistically upregulated the mRNA expression of IL-6 and CXC chemokines (Figure 3B).

Bottom Line: The mRNA expression of proinflammatory mediators induced by IL-36α and/or IL-36γ was significantly suppressed by transfection of siRNA for MyD88 or IRAK1.Both inhibitors of mitogen activated protein kinases and siRNAs specific for NF-κBp65 significantly reduced the expression of IL-6 and CXC chemokines induced by IL-36α and/or IL-36γ.These results suggest that IL-36α and IL-36γ contribute to gut inflammation through the induction of proinflammatory mediators.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Shiga University of Medical Science , Otsu , Japan.

ABSTRACT

Background: Interleukin (IL)-36 cytokines are recently reported member of the IL-1 cytokine family. However, there is little information regarding the association between IL-36 cytokines and gut inflammation. In the present study, we investigated the biological activity of IL-36α and IL-36γ using human colonic subepithelial myofibroblasts (SEMFs).

Methods: The mRNA expression and the protein expression of target molecules in SEMFs were evaluated using real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The intracellular signaling of IL-36 cytokines was analyzed using Western blot analysis and small interfering RNAs (siRNAs) specific for MyD88 adaptor proteins (MyD88 and IRAK1) and NF-κB p65.

Results: IL-36α and IL-36γ significantly enhanced the secretion of IL-6 and CXC chemokines (CXCL1, CXCL2, and CXCL8) by SEMFs. The combination of IL-36α/γ and IL-17A or of IL-36α/γ and tumor necrosis factor-α showed a synergistic effect on the induction of IL-6 and CXC chemokines. The mRNA expression of proinflammatory mediators induced by IL-36α and/or IL-36γ was significantly suppressed by transfection of siRNA for MyD88 or IRAK1. Both inhibitors of mitogen activated protein kinases and siRNAs specific for NF-κBp65 significantly reduced the expression of IL-6 and CXC chemokines induced by IL-36α and/or IL-36γ.

Conclusion: These results suggest that IL-36α and IL-36γ contribute to gut inflammation through the induction of proinflammatory mediators.

No MeSH data available.


Related in: MedlinePlus