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Synthetic RGDS peptide attenuates lipopolysaccharide-induced pulmonary inflammation by inhibiting integrin signaled MAP kinase pathways.

Moon C, Han JR, Park HJ, Hah JS, Kang JL - Respir. Res. (2009)

Bottom Line: RGDS inhibited LPS-induced phosphorylation of focal adhesion kinase and MAP kinases, including ERK, JNK, and p38 MAP kinase, in lung tissue.Similarly, a blocking antibody against integrin alphav significantly inhibited LPS-induced inflammatory cell migration into the lung, protein accumulation and proinflammatory mediator production in BAL fluid, at 4 or 24 h post-LPS.These results suggest that RGDS with high specificity for alphavintegrins attenuates inflammatory cascade during LPS-induced development of acute lung injury.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Physiology, Division of Cell Biology, Ewha Medical Research Center, School of Medicine, Ewha Womans University, Seoul, Seoul, Korea. 94cmoon@hanmail.net

ABSTRACT

Background: Synthetic peptides containing the RGD sequence inhibit integrin-related functions in different cell systems. Here, we investigated the effects of synthetic Arg-Gly-Asp-Ser (RGDS) peptide on key inflammatory responses to intratracheal (i.t.) lipopolysaccharide (LPS) treatment and on the integrin signaled mitogen-activated protein (MAP) kinase pathway during the development of acute lung injury.

Methods: Saline or LPS (1.5 mg/kg) was administered i.t. with or without a single dose of RGDS (1, 2.5, or 5 mg/kg, i.p.), anti-alphav or anti-beta3 mAb (5 mg/kg, i.p.). Mice were sacrificed 4 or 24 h post-LPS.

Results: A pretreatment with RGDS inhibited LPS-induced increases in neutrophil and macrophage numbers, total protein levels and TNF-alpha and MIP-2 levels, and matrix metalloproteinase-9 activity in bronchoalveolar lavage (BAL) fluid at 4 or 24 h post-LPS treatment. RGDS inhibited LPS-induced phosphorylation of focal adhesion kinase and MAP kinases, including ERK, JNK, and p38 MAP kinase, in lung tissue. Importantly, the inhibition of the inflammatory responses and the kinase pathways were still evident when this peptide was administered 2 h after LPS treatment. Similarly, a blocking antibody against integrin alphav significantly inhibited LPS-induced inflammatory cell migration into the lung, protein accumulation and proinflammatory mediator production in BAL fluid, at 4 or 24 h post-LPS. Anti-beta3 also inhibited all LPS-induced inflammatory responses, except the accumulation of BAL protein at 24 h post-LPS.

Conclusion: These results suggest that RGDS with high specificity for alphavintegrins attenuates inflammatory cascade during LPS-induced development of acute lung injury.

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Effects of RGDS on phosphorylation of ERK1/2 (A, B), JNK1/2(C, D) and p38 MAP kinase (E, F) in lung tissue. Where indicated, mice were administered RGDS or RGES (5 mg/kg, i.p.) once 1 h before LPS treatment and sacrificed 4 h post-LPS. Mice were also administered these peptides (5 mg/kg, i.p.) once 1 h before or 2 h after LPS and sacrificed 24 h post-LPS. Western blotting was performed with each anti-specific (phospho) Ab on lung tissue homogenates. Relative values for phosphorylated MAP kinase versus MAP kinase, respectively, are indicated below the gel. Values represent means ± SEM of 5 mice per group. *Significantly different from saline treated controls, p < 0.05; +significantly different from animals treated with LPS only, p < 0.05.
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Figure 6: Effects of RGDS on phosphorylation of ERK1/2 (A, B), JNK1/2(C, D) and p38 MAP kinase (E, F) in lung tissue. Where indicated, mice were administered RGDS or RGES (5 mg/kg, i.p.) once 1 h before LPS treatment and sacrificed 4 h post-LPS. Mice were also administered these peptides (5 mg/kg, i.p.) once 1 h before or 2 h after LPS and sacrificed 24 h post-LPS. Western blotting was performed with each anti-specific (phospho) Ab on lung tissue homogenates. Relative values for phosphorylated MAP kinase versus MAP kinase, respectively, are indicated below the gel. Values represent means ± SEM of 5 mice per group. *Significantly different from saline treated controls, p < 0.05; +significantly different from animals treated with LPS only, p < 0.05.

Mentions: Figure 6 presents LPS-induced MAP kinase phosphorylation data in lung tissue at 4 or 24 h post-LPS, which concurs with the findings of our previous study [28,29]. Phosphorylation of ERK and JNK substantially were increased 4 h post-LPS, and were either maintained (ERK) (Figures 6A and 6B) or further increased (JNK) (Figures 6C and 6D). p38 MAP kinase phosphorylation substantially increased at 4 h and this phosphorylation level was slightly reduced 24 h post-LPS (Figures 6E and 6F). RGDS treatment 1 h before LPS significantly suppressed LPS-induced phosphorylation of ERK, JNK and p38 MAP kinase 4 h post-LPS. Pre- or posttreatment with RGDS also inhibited all three MAP kinase phosphorylation 24 h post-LPS. The smallest decrease was observed for p38 MAP kinase. These findings suggest that RGD motif-dependent integrin signaling, involving FAK activation, is solidly linked to LPS-induced ERK and JNK activations during the progression of acute lung injury, and that it is weakly linked to p38 MAP kinase activation.


Synthetic RGDS peptide attenuates lipopolysaccharide-induced pulmonary inflammation by inhibiting integrin signaled MAP kinase pathways.

Moon C, Han JR, Park HJ, Hah JS, Kang JL - Respir. Res. (2009)

Effects of RGDS on phosphorylation of ERK1/2 (A, B), JNK1/2(C, D) and p38 MAP kinase (E, F) in lung tissue. Where indicated, mice were administered RGDS or RGES (5 mg/kg, i.p.) once 1 h before LPS treatment and sacrificed 4 h post-LPS. Mice were also administered these peptides (5 mg/kg, i.p.) once 1 h before or 2 h after LPS and sacrificed 24 h post-LPS. Western blotting was performed with each anti-specific (phospho) Ab on lung tissue homogenates. Relative values for phosphorylated MAP kinase versus MAP kinase, respectively, are indicated below the gel. Values represent means ± SEM of 5 mice per group. *Significantly different from saline treated controls, p < 0.05; +significantly different from animals treated with LPS only, p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 6: Effects of RGDS on phosphorylation of ERK1/2 (A, B), JNK1/2(C, D) and p38 MAP kinase (E, F) in lung tissue. Where indicated, mice were administered RGDS or RGES (5 mg/kg, i.p.) once 1 h before LPS treatment and sacrificed 4 h post-LPS. Mice were also administered these peptides (5 mg/kg, i.p.) once 1 h before or 2 h after LPS and sacrificed 24 h post-LPS. Western blotting was performed with each anti-specific (phospho) Ab on lung tissue homogenates. Relative values for phosphorylated MAP kinase versus MAP kinase, respectively, are indicated below the gel. Values represent means ± SEM of 5 mice per group. *Significantly different from saline treated controls, p < 0.05; +significantly different from animals treated with LPS only, p < 0.05.
Mentions: Figure 6 presents LPS-induced MAP kinase phosphorylation data in lung tissue at 4 or 24 h post-LPS, which concurs with the findings of our previous study [28,29]. Phosphorylation of ERK and JNK substantially were increased 4 h post-LPS, and were either maintained (ERK) (Figures 6A and 6B) or further increased (JNK) (Figures 6C and 6D). p38 MAP kinase phosphorylation substantially increased at 4 h and this phosphorylation level was slightly reduced 24 h post-LPS (Figures 6E and 6F). RGDS treatment 1 h before LPS significantly suppressed LPS-induced phosphorylation of ERK, JNK and p38 MAP kinase 4 h post-LPS. Pre- or posttreatment with RGDS also inhibited all three MAP kinase phosphorylation 24 h post-LPS. The smallest decrease was observed for p38 MAP kinase. These findings suggest that RGD motif-dependent integrin signaling, involving FAK activation, is solidly linked to LPS-induced ERK and JNK activations during the progression of acute lung injury, and that it is weakly linked to p38 MAP kinase activation.

Bottom Line: RGDS inhibited LPS-induced phosphorylation of focal adhesion kinase and MAP kinases, including ERK, JNK, and p38 MAP kinase, in lung tissue.Similarly, a blocking antibody against integrin alphav significantly inhibited LPS-induced inflammatory cell migration into the lung, protein accumulation and proinflammatory mediator production in BAL fluid, at 4 or 24 h post-LPS.These results suggest that RGDS with high specificity for alphavintegrins attenuates inflammatory cascade during LPS-induced development of acute lung injury.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Physiology, Division of Cell Biology, Ewha Medical Research Center, School of Medicine, Ewha Womans University, Seoul, Seoul, Korea. 94cmoon@hanmail.net

ABSTRACT

Background: Synthetic peptides containing the RGD sequence inhibit integrin-related functions in different cell systems. Here, we investigated the effects of synthetic Arg-Gly-Asp-Ser (RGDS) peptide on key inflammatory responses to intratracheal (i.t.) lipopolysaccharide (LPS) treatment and on the integrin signaled mitogen-activated protein (MAP) kinase pathway during the development of acute lung injury.

Methods: Saline or LPS (1.5 mg/kg) was administered i.t. with or without a single dose of RGDS (1, 2.5, or 5 mg/kg, i.p.), anti-alphav or anti-beta3 mAb (5 mg/kg, i.p.). Mice were sacrificed 4 or 24 h post-LPS.

Results: A pretreatment with RGDS inhibited LPS-induced increases in neutrophil and macrophage numbers, total protein levels and TNF-alpha and MIP-2 levels, and matrix metalloproteinase-9 activity in bronchoalveolar lavage (BAL) fluid at 4 or 24 h post-LPS treatment. RGDS inhibited LPS-induced phosphorylation of focal adhesion kinase and MAP kinases, including ERK, JNK, and p38 MAP kinase, in lung tissue. Importantly, the inhibition of the inflammatory responses and the kinase pathways were still evident when this peptide was administered 2 h after LPS treatment. Similarly, a blocking antibody against integrin alphav significantly inhibited LPS-induced inflammatory cell migration into the lung, protein accumulation and proinflammatory mediator production in BAL fluid, at 4 or 24 h post-LPS. Anti-beta3 also inhibited all LPS-induced inflammatory responses, except the accumulation of BAL protein at 24 h post-LPS.

Conclusion: These results suggest that RGDS with high specificity for alphavintegrins attenuates inflammatory cascade during LPS-induced development of acute lung injury.

Show MeSH
Related in: MedlinePlus