Limits...
The selective phosphodiesterase 4 inhibitor roflumilast and phosphodiesterase 3/4 inhibitor pumafentrine reduce clinical score and TNF expression in experimental colitis in mice.

Rieder F, Siegmund B, Bundschuh DS, Lehr HA, Endres S, Eigler A - PLoS ONE (2013)

Bottom Line: Roflumilast treatment resulted in dose-dependent improvements of clinical score (weight loss, stool consistency and bleeding), colon length, and local tumor necrosis factor-α (TNFα) production in the colonic tissue.These findings, however, were not associated with an improvement of the histologic score.In vitro stimulated splenocytes after in vivo treatment with pumafentrine showed a significantly lower state of activation and production of IFNγ compared to no treatment in vivo.

View Article: PubMed Central - PubMed

Affiliation: Division of Clinical Pharmacology and Section of Gastroenterology, University of Munich, Munich, Germany.

ABSTRACT

Objective: The specific inhibition of phosphodiesterase (PDE)4 and dual inhibition of PDE3 and PDE4 has been shown to decrease inflammation by suppression of pro-inflammatory cytokine synthesis. We examined the effect of roflumilast, a selective PDE4 inhibitor marketed for severe COPD, and the investigational compound pumafentrine, a dual PDE3/PDE4 inhibitor, in the preventive dextran sodium sulfate (DSS)-induced colitis model.

Methods: The clinical score, colon length, histologic score and colon cytokine production from mice with DSS-induced colitis (3.5% DSS in drinking water for 11 days) receiving either roflumilast (1 or 5 mg/kg body weight/d p.o.) or pumafentrine (1.5 or 5 mg/kg/d p.o.) were determined and compared to vehicle treated control mice. In the pumafentrine-treated animals, splenocytes were analyzed for interferon-γ (IFNγ) production and CD69 expression.

Results: Roflumilast treatment resulted in dose-dependent improvements of clinical score (weight loss, stool consistency and bleeding), colon length, and local tumor necrosis factor-α (TNFα) production in the colonic tissue. These findings, however, were not associated with an improvement of the histologic score. Administration of pumafentrine at 5 mg/kg/d alleviated the clinical score, the colon length shortening, and local TNFα production. In vitro stimulated splenocytes after in vivo treatment with pumafentrine showed a significantly lower state of activation and production of IFNγ compared to no treatment in vivo.

Conclusions: These series of experiments document the ameliorating effect of roflumilast and pumafentrine on the clinical score and TNF expression of experimental colitis in mice.

Show MeSH

Related in: MedlinePlus

Effect of pumafentrine on clinical score, colon length and histologic score. A. Efficacy of pumafentrine in DSS-induced colitis.Mice were exposed to 3.5% DSS in drinking water for eleven days. Either 1.5 mg/kg/d pumafentrine (n = 8), 5 mg/kg/d pumafentrine (n = 16) or 4% methocel (n = 16) were administered orally once daily for eleven days. Non-DSS mice received either 20 mg/kg/d pumafentrine or 4% methocel (n = 8). The degree of colitis was quantified by the clinical score assessing weight loss, stool consistency and rectal bleeding (range from 0 =  healthy to 4 =  maximal disease activity). The scores are depicted as mean ± SEM; *p<0.05, **p<0.01 versus DSS+methocel. B. Reduction of colon length shortening in DSS-induced colitis by pumafentrine. Mice were exposed to 3.5% DSS in drinking water for an eleven day period. Pumafentrine treatment (either 1.5 mg/kg/d (n = 8) or 5 mg/kg/d (n = 16) orally, once daily for 11 days) or 4% methocel (n = 12) were started the same day as DSS administration. Non-DSS mice received 20 mg/kg/d pumafentrine or 4% methocel (n = 8), respectively. Values are depicted as mean ± SEM. **p<0.01 versus DSS+methocel. C. Effect on histologic signs of colonic inflammation by pumafentrine. Mice were exposed to 3.5% DSS in drinking water for eleven days and were treated with pumafentrine (either 1.5 mg/kg/d (n = 8) or 5 mg/kg/d (n = 16) orally once daily for 11 days) or 4% methocel (n = 12). Non-DSS mice received 20 mg/kg/d pumafentrine or 4% methocel (n = 8). At day 11 mice were euthanized, colon rings were stained and the histologic score (degree of inflammation: 0 =  no changes to 6 =  extensive cell infiltration and tissue damage) was determined in a blinded fashion as described in detail in the Material and Methods. Scores are depicted as means ± SEM.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3585290&req=5

pone-0056867-g004: Effect of pumafentrine on clinical score, colon length and histologic score. A. Efficacy of pumafentrine in DSS-induced colitis.Mice were exposed to 3.5% DSS in drinking water for eleven days. Either 1.5 mg/kg/d pumafentrine (n = 8), 5 mg/kg/d pumafentrine (n = 16) or 4% methocel (n = 16) were administered orally once daily for eleven days. Non-DSS mice received either 20 mg/kg/d pumafentrine or 4% methocel (n = 8). The degree of colitis was quantified by the clinical score assessing weight loss, stool consistency and rectal bleeding (range from 0 =  healthy to 4 =  maximal disease activity). The scores are depicted as mean ± SEM; *p<0.05, **p<0.01 versus DSS+methocel. B. Reduction of colon length shortening in DSS-induced colitis by pumafentrine. Mice were exposed to 3.5% DSS in drinking water for an eleven day period. Pumafentrine treatment (either 1.5 mg/kg/d (n = 8) or 5 mg/kg/d (n = 16) orally, once daily for 11 days) or 4% methocel (n = 12) were started the same day as DSS administration. Non-DSS mice received 20 mg/kg/d pumafentrine or 4% methocel (n = 8), respectively. Values are depicted as mean ± SEM. **p<0.01 versus DSS+methocel. C. Effect on histologic signs of colonic inflammation by pumafentrine. Mice were exposed to 3.5% DSS in drinking water for eleven days and were treated with pumafentrine (either 1.5 mg/kg/d (n = 8) or 5 mg/kg/d (n = 16) orally once daily for 11 days) or 4% methocel (n = 12). Non-DSS mice received 20 mg/kg/d pumafentrine or 4% methocel (n = 8). At day 11 mice were euthanized, colon rings were stained and the histologic score (degree of inflammation: 0 =  no changes to 6 =  extensive cell infiltration and tissue damage) was determined in a blinded fashion as described in detail in the Material and Methods. Scores are depicted as means ± SEM.

Mentions: Mice exposed to 3.5% DSS developed signs of colitis as expressed by a clinical score of greater than 0.5 from day 4 onward (Figure 4A). Oral administration of pumafentrine 5 mg/kg/d retarded the onset of colitis to day 6. No dose dependency of the treatment was noted starting day 8. Treatment with pumafentrine 5 mg/kg/d resulted in a lower clinical score starting at day 6 of the experiment and lasting until the end of the experiment on day 11 (n = 16; Figure 4A). Administration of 1.5 mg/kg/d pumafentrine did not delay the onset of clinical signs of colitis and also did not influence the clinical score until day 11 when compared to the untreated DSS animals. The non-DSS control animals, which received regular drinking water and pumafentrine 5 mg/kg/d, 20 mg/kg/d, or 4% methocel (n = 8) did not develop signs of colitis during the experimental course (all clinical scores <0.5 on days 1 to 11).


The selective phosphodiesterase 4 inhibitor roflumilast and phosphodiesterase 3/4 inhibitor pumafentrine reduce clinical score and TNF expression in experimental colitis in mice.

Rieder F, Siegmund B, Bundschuh DS, Lehr HA, Endres S, Eigler A - PLoS ONE (2013)

Effect of pumafentrine on clinical score, colon length and histologic score. A. Efficacy of pumafentrine in DSS-induced colitis.Mice were exposed to 3.5% DSS in drinking water for eleven days. Either 1.5 mg/kg/d pumafentrine (n = 8), 5 mg/kg/d pumafentrine (n = 16) or 4% methocel (n = 16) were administered orally once daily for eleven days. Non-DSS mice received either 20 mg/kg/d pumafentrine or 4% methocel (n = 8). The degree of colitis was quantified by the clinical score assessing weight loss, stool consistency and rectal bleeding (range from 0 =  healthy to 4 =  maximal disease activity). The scores are depicted as mean ± SEM; *p<0.05, **p<0.01 versus DSS+methocel. B. Reduction of colon length shortening in DSS-induced colitis by pumafentrine. Mice were exposed to 3.5% DSS in drinking water for an eleven day period. Pumafentrine treatment (either 1.5 mg/kg/d (n = 8) or 5 mg/kg/d (n = 16) orally, once daily for 11 days) or 4% methocel (n = 12) were started the same day as DSS administration. Non-DSS mice received 20 mg/kg/d pumafentrine or 4% methocel (n = 8), respectively. Values are depicted as mean ± SEM. **p<0.01 versus DSS+methocel. C. Effect on histologic signs of colonic inflammation by pumafentrine. Mice were exposed to 3.5% DSS in drinking water for eleven days and were treated with pumafentrine (either 1.5 mg/kg/d (n = 8) or 5 mg/kg/d (n = 16) orally once daily for 11 days) or 4% methocel (n = 12). Non-DSS mice received 20 mg/kg/d pumafentrine or 4% methocel (n = 8). At day 11 mice were euthanized, colon rings were stained and the histologic score (degree of inflammation: 0 =  no changes to 6 =  extensive cell infiltration and tissue damage) was determined in a blinded fashion as described in detail in the Material and Methods. Scores are depicted as means ± SEM.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585290&req=5

pone-0056867-g004: Effect of pumafentrine on clinical score, colon length and histologic score. A. Efficacy of pumafentrine in DSS-induced colitis.Mice were exposed to 3.5% DSS in drinking water for eleven days. Either 1.5 mg/kg/d pumafentrine (n = 8), 5 mg/kg/d pumafentrine (n = 16) or 4% methocel (n = 16) were administered orally once daily for eleven days. Non-DSS mice received either 20 mg/kg/d pumafentrine or 4% methocel (n = 8). The degree of colitis was quantified by the clinical score assessing weight loss, stool consistency and rectal bleeding (range from 0 =  healthy to 4 =  maximal disease activity). The scores are depicted as mean ± SEM; *p<0.05, **p<0.01 versus DSS+methocel. B. Reduction of colon length shortening in DSS-induced colitis by pumafentrine. Mice were exposed to 3.5% DSS in drinking water for an eleven day period. Pumafentrine treatment (either 1.5 mg/kg/d (n = 8) or 5 mg/kg/d (n = 16) orally, once daily for 11 days) or 4% methocel (n = 12) were started the same day as DSS administration. Non-DSS mice received 20 mg/kg/d pumafentrine or 4% methocel (n = 8), respectively. Values are depicted as mean ± SEM. **p<0.01 versus DSS+methocel. C. Effect on histologic signs of colonic inflammation by pumafentrine. Mice were exposed to 3.5% DSS in drinking water for eleven days and were treated with pumafentrine (either 1.5 mg/kg/d (n = 8) or 5 mg/kg/d (n = 16) orally once daily for 11 days) or 4% methocel (n = 12). Non-DSS mice received 20 mg/kg/d pumafentrine or 4% methocel (n = 8). At day 11 mice were euthanized, colon rings were stained and the histologic score (degree of inflammation: 0 =  no changes to 6 =  extensive cell infiltration and tissue damage) was determined in a blinded fashion as described in detail in the Material and Methods. Scores are depicted as means ± SEM.
Mentions: Mice exposed to 3.5% DSS developed signs of colitis as expressed by a clinical score of greater than 0.5 from day 4 onward (Figure 4A). Oral administration of pumafentrine 5 mg/kg/d retarded the onset of colitis to day 6. No dose dependency of the treatment was noted starting day 8. Treatment with pumafentrine 5 mg/kg/d resulted in a lower clinical score starting at day 6 of the experiment and lasting until the end of the experiment on day 11 (n = 16; Figure 4A). Administration of 1.5 mg/kg/d pumafentrine did not delay the onset of clinical signs of colitis and also did not influence the clinical score until day 11 when compared to the untreated DSS animals. The non-DSS control animals, which received regular drinking water and pumafentrine 5 mg/kg/d, 20 mg/kg/d, or 4% methocel (n = 8) did not develop signs of colitis during the experimental course (all clinical scores <0.5 on days 1 to 11).

Bottom Line: Roflumilast treatment resulted in dose-dependent improvements of clinical score (weight loss, stool consistency and bleeding), colon length, and local tumor necrosis factor-α (TNFα) production in the colonic tissue.These findings, however, were not associated with an improvement of the histologic score.In vitro stimulated splenocytes after in vivo treatment with pumafentrine showed a significantly lower state of activation and production of IFNγ compared to no treatment in vivo.

View Article: PubMed Central - PubMed

Affiliation: Division of Clinical Pharmacology and Section of Gastroenterology, University of Munich, Munich, Germany.

ABSTRACT

Objective: The specific inhibition of phosphodiesterase (PDE)4 and dual inhibition of PDE3 and PDE4 has been shown to decrease inflammation by suppression of pro-inflammatory cytokine synthesis. We examined the effect of roflumilast, a selective PDE4 inhibitor marketed for severe COPD, and the investigational compound pumafentrine, a dual PDE3/PDE4 inhibitor, in the preventive dextran sodium sulfate (DSS)-induced colitis model.

Methods: The clinical score, colon length, histologic score and colon cytokine production from mice with DSS-induced colitis (3.5% DSS in drinking water for 11 days) receiving either roflumilast (1 or 5 mg/kg body weight/d p.o.) or pumafentrine (1.5 or 5 mg/kg/d p.o.) were determined and compared to vehicle treated control mice. In the pumafentrine-treated animals, splenocytes were analyzed for interferon-γ (IFNγ) production and CD69 expression.

Results: Roflumilast treatment resulted in dose-dependent improvements of clinical score (weight loss, stool consistency and bleeding), colon length, and local tumor necrosis factor-α (TNFα) production in the colonic tissue. These findings, however, were not associated with an improvement of the histologic score. Administration of pumafentrine at 5 mg/kg/d alleviated the clinical score, the colon length shortening, and local TNFα production. In vitro stimulated splenocytes after in vivo treatment with pumafentrine showed a significantly lower state of activation and production of IFNγ compared to no treatment in vivo.

Conclusions: These series of experiments document the ameliorating effect of roflumilast and pumafentrine on the clinical score and TNF expression of experimental colitis in mice.

Show MeSH
Related in: MedlinePlus