Limits...
Lovastatin Inhibits Low Molecular Weight Hyaluronan Induced Chemokine Expression via LFA-1 and Decreases Bleomycin-Induced Pulmonary Fibrosis.

Hamblin MJ, Eberlein M, Black K, Hallowell R, Collins S, Chan-Li Y, Horton MR - Int J Biomed Sci (2014)

Bottom Line: We evaluated the effects of lovastatin, pravastatin, LFA-1 blocking antibodies, and a novel LFA-1 inhibitor LFA 878 on LMW HA induced cytokine production in alveolar macrophages.Lovastatin immediately inhibited the LMW HA induced cytokine MIP 1-α (p=0.001) independent of HMG CoA reductase.Lovastatin and LFA 878 inhibit LMW HA inflammatory signaling independent of HMG-CoA decreasing the chemotactic cytokine MIP 1-α.

View Article: PubMed Central - PubMed

Affiliation: University of Kansas Hospital, USA;

ABSTRACT

Background: Lovastatin has a unique ability to bind Leukocyte Function Antigen-1 (LFA-1), an integrin necessary for the full expression of inflammatory cytokines induced by the low molecular weight form of the extracellular matrix glycosaminoglycan hyaluronan (LMW HA). We hypothesized that lovastatin could inhibit LMW HA inflammatory signals via interaction with LFA-1, and attenuate bleomycin induced pulmonary fibrosis.

Methods: We evaluated the effects of lovastatin, pravastatin, LFA-1 blocking antibodies, and a novel LFA-1 inhibitor LFA 878 on LMW HA induced cytokine production in alveolar macrophages. We evaluated the effect of lovastatin in a bleomycin model of lung injury.

Results: Lovastatin immediately inhibited the LMW HA induced cytokine MIP 1-α (p=0.001) independent of HMG CoA reductase. Pravastatin showed no inhibitory profile when administered simultaneously with LMW HA. LFA-1 blocking antibodies and the small molecule statin derivative LFA 878 showed an inhibitory profile similar to lovastatin. Lovastatin showed decreased fibrosis on histopathology and improved survival at day 14, with a decrease in fibrocytes noted at day 8.

Conclusion: Lovastatin and LFA 878 inhibit LMW HA inflammatory signaling independent of HMG-CoA decreasing the chemotactic cytokine MIP 1-α. Lovastatin treatment improves survival in bleomycin lung injury with decreased fibrocytes and fibrosis.

No MeSH data available.


Related in: MedlinePlus

A, MIP 1-α protein by ELISA in MH-S alveolar macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and varying doses of lovastatin (0, 1, 5, 10, 20, 30, and 40 μM). Lovastatin significantly inhibited MIP 1-α production with an IC50 noted at a dose of 5 μM (p=0.001); B, MIP 1-α protein by ELISA in Thioglycollate derived peritoneal macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and lovastatin (20 μM). Lovastatin significantly inhibited MIP 1-α production; C-E, KC, RANTES and TNF-α protein by ELISA in MH-S alveolar macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and lovastatin (20 μM). Lovastatin significantly inhibited KC (p=0.005), and RANTES (0.009) but not TNF-α expression; F, MH-S Cells were stimulated with vehicle, LMW HA (250 μg/ml) alone or with simultaneous administration of lovastatin (20 μM) or lovastatin (20 μM) and mevonalate (500 μM). MIP 1-α mRNA expression was measured by Northern Blot analysis. Lovastatin significantly inhibited LMW HA induction of MIP 1-α DNA, and the addition of mevonalate did not restore MIP 1-α mRNA expression.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4199473&req=5

Figure 2: A, MIP 1-α protein by ELISA in MH-S alveolar macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and varying doses of lovastatin (0, 1, 5, 10, 20, 30, and 40 μM). Lovastatin significantly inhibited MIP 1-α production with an IC50 noted at a dose of 5 μM (p=0.001); B, MIP 1-α protein by ELISA in Thioglycollate derived peritoneal macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and lovastatin (20 μM). Lovastatin significantly inhibited MIP 1-α production; C-E, KC, RANTES and TNF-α protein by ELISA in MH-S alveolar macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and lovastatin (20 μM). Lovastatin significantly inhibited KC (p=0.005), and RANTES (0.009) but not TNF-α expression; F, MH-S Cells were stimulated with vehicle, LMW HA (250 μg/ml) alone or with simultaneous administration of lovastatin (20 μM) or lovastatin (20 μM) and mevonalate (500 μM). MIP 1-α mRNA expression was measured by Northern Blot analysis. Lovastatin significantly inhibited LMW HA induction of MIP 1-α DNA, and the addition of mevonalate did not restore MIP 1-α mRNA expression.

Mentions: Experiments were carried out in both wild type (WT) murine peritoneal derived macrophages and the MH-S cell line of murine alveolar macrophages. We initially determined the optimal inhibitory dose of lovastatin by stimulating MH-S alveolar macrophages with increasing doses of lovastatin ranging from 1 uM to 40 uM. Lovastatin was added to cell culture immediately prior to LMW HA (250 ug/ml) administration. MIP 1-α cytokine production was selected for primary analysis, because activation and cross-linking of LFA-1 has been shown to induce secretion of MIP 1-a leading to lymphocyte migration to areas of inflammation (27) . Protein supernatants were collected at 18 hours, and analyzed by ELISA for MIP 1-α cytokine production. We found that LMW HA markedly up-regulated MIP 1-α, and lovastatin inhibited this production in a dose dependent manner (Figure 2A). The IC50 appears to occur at a dose of 5 uM, but maximal inhibition occurred at a dose of 40 uM. Preliminary experiments demonstrated doses greater than 40 uM resulted in excessive cell death (>5%). To avoid a potential confounding effect a 20 uM dose (p=0.001) was determined to achieve a near maximal inhibition while avoiding the potential cytotoxic effects of higher doses.


Lovastatin Inhibits Low Molecular Weight Hyaluronan Induced Chemokine Expression via LFA-1 and Decreases Bleomycin-Induced Pulmonary Fibrosis.

Hamblin MJ, Eberlein M, Black K, Hallowell R, Collins S, Chan-Li Y, Horton MR - Int J Biomed Sci (2014)

A, MIP 1-α protein by ELISA in MH-S alveolar macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and varying doses of lovastatin (0, 1, 5, 10, 20, 30, and 40 μM). Lovastatin significantly inhibited MIP 1-α production with an IC50 noted at a dose of 5 μM (p=0.001); B, MIP 1-α protein by ELISA in Thioglycollate derived peritoneal macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and lovastatin (20 μM). Lovastatin significantly inhibited MIP 1-α production; C-E, KC, RANTES and TNF-α protein by ELISA in MH-S alveolar macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and lovastatin (20 μM). Lovastatin significantly inhibited KC (p=0.005), and RANTES (0.009) but not TNF-α expression; F, MH-S Cells were stimulated with vehicle, LMW HA (250 μg/ml) alone or with simultaneous administration of lovastatin (20 μM) or lovastatin (20 μM) and mevonalate (500 μM). MIP 1-α mRNA expression was measured by Northern Blot analysis. Lovastatin significantly inhibited LMW HA induction of MIP 1-α DNA, and the addition of mevonalate did not restore MIP 1-α mRNA expression.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4199473&req=5

Figure 2: A, MIP 1-α protein by ELISA in MH-S alveolar macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and varying doses of lovastatin (0, 1, 5, 10, 20, 30, and 40 μM). Lovastatin significantly inhibited MIP 1-α production with an IC50 noted at a dose of 5 μM (p=0.001); B, MIP 1-α protein by ELISA in Thioglycollate derived peritoneal macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and lovastatin (20 μM). Lovastatin significantly inhibited MIP 1-α production; C-E, KC, RANTES and TNF-α protein by ELISA in MH-S alveolar macrophages treated with simultaneous administration of LMW HA (250 μg/ml) and lovastatin (20 μM). Lovastatin significantly inhibited KC (p=0.005), and RANTES (0.009) but not TNF-α expression; F, MH-S Cells were stimulated with vehicle, LMW HA (250 μg/ml) alone or with simultaneous administration of lovastatin (20 μM) or lovastatin (20 μM) and mevonalate (500 μM). MIP 1-α mRNA expression was measured by Northern Blot analysis. Lovastatin significantly inhibited LMW HA induction of MIP 1-α DNA, and the addition of mevonalate did not restore MIP 1-α mRNA expression.
Mentions: Experiments were carried out in both wild type (WT) murine peritoneal derived macrophages and the MH-S cell line of murine alveolar macrophages. We initially determined the optimal inhibitory dose of lovastatin by stimulating MH-S alveolar macrophages with increasing doses of lovastatin ranging from 1 uM to 40 uM. Lovastatin was added to cell culture immediately prior to LMW HA (250 ug/ml) administration. MIP 1-α cytokine production was selected for primary analysis, because activation and cross-linking of LFA-1 has been shown to induce secretion of MIP 1-a leading to lymphocyte migration to areas of inflammation (27) . Protein supernatants were collected at 18 hours, and analyzed by ELISA for MIP 1-α cytokine production. We found that LMW HA markedly up-regulated MIP 1-α, and lovastatin inhibited this production in a dose dependent manner (Figure 2A). The IC50 appears to occur at a dose of 5 uM, but maximal inhibition occurred at a dose of 40 uM. Preliminary experiments demonstrated doses greater than 40 uM resulted in excessive cell death (>5%). To avoid a potential confounding effect a 20 uM dose (p=0.001) was determined to achieve a near maximal inhibition while avoiding the potential cytotoxic effects of higher doses.

Bottom Line: We evaluated the effects of lovastatin, pravastatin, LFA-1 blocking antibodies, and a novel LFA-1 inhibitor LFA 878 on LMW HA induced cytokine production in alveolar macrophages.Lovastatin immediately inhibited the LMW HA induced cytokine MIP 1-α (p=0.001) independent of HMG CoA reductase.Lovastatin and LFA 878 inhibit LMW HA inflammatory signaling independent of HMG-CoA decreasing the chemotactic cytokine MIP 1-α.

View Article: PubMed Central - PubMed

Affiliation: University of Kansas Hospital, USA;

ABSTRACT

Background: Lovastatin has a unique ability to bind Leukocyte Function Antigen-1 (LFA-1), an integrin necessary for the full expression of inflammatory cytokines induced by the low molecular weight form of the extracellular matrix glycosaminoglycan hyaluronan (LMW HA). We hypothesized that lovastatin could inhibit LMW HA inflammatory signals via interaction with LFA-1, and attenuate bleomycin induced pulmonary fibrosis.

Methods: We evaluated the effects of lovastatin, pravastatin, LFA-1 blocking antibodies, and a novel LFA-1 inhibitor LFA 878 on LMW HA induced cytokine production in alveolar macrophages. We evaluated the effect of lovastatin in a bleomycin model of lung injury.

Results: Lovastatin immediately inhibited the LMW HA induced cytokine MIP 1-α (p=0.001) independent of HMG CoA reductase. Pravastatin showed no inhibitory profile when administered simultaneously with LMW HA. LFA-1 blocking antibodies and the small molecule statin derivative LFA 878 showed an inhibitory profile similar to lovastatin. Lovastatin showed decreased fibrosis on histopathology and improved survival at day 14, with a decrease in fibrocytes noted at day 8.

Conclusion: Lovastatin and LFA 878 inhibit LMW HA inflammatory signaling independent of HMG-CoA decreasing the chemotactic cytokine MIP 1-α. Lovastatin treatment improves survival in bleomycin lung injury with decreased fibrocytes and fibrosis.

No MeSH data available.


Related in: MedlinePlus