PMC

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.

Mice were exposed to 3.5% DSS in drinking water for 11 days and were treated with pumafentrine 5 mg/kg/d (n = 8) orally once daily for 11 days) or 4% methocel (n = 8). Non-DSS mice received 20 mg/kg/d pumafentrine (n = 8). After day 11 spleens were removed aseptically and splenocytes were isolated. After a 20 h incubation period with PMA 25 ng/ml and ionomycine 500 ng/ml, the experiment was stopped by three freeze-thaw cycles and total IFNγ was measured by ELISA. Results are depicted as mean ± SEM. *p<0.05 versus DSS+methocel. B. Effect of in vivo administered pumafentrine on PMA/ionomycine-induced CD69 expression by splenocytes from DSS-exposed mice. Mice were exposed to 3.5% DSS in drinking water for 11 days and were treated with pumafentrine 5 mg/kg/d (n = 8) orally once daily for 11 days) or 4% methocel (n = 8). Non-DSS mice received 20 mg/kg/d pumafentrine (n = 8). After day 11 spleens were removed aseptically and splenocytes were isolated and stimulated with PMA 25 ng/ml and ionomycine 500 ng/ml. After 20 h incubation period the CD69-expression was determined by flow cytometry. Bars represent the mean ± SEM of CD69 positive splenocytes. *p<0.05 versus DSS+methocel.

Mice were exposed to 3.5% DSS in drinking water for 11 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, the end of experiment, the colon was removed, weighed, vortexed in PBS and centrifuged. TNFα was quantified in the eluate by ELISA. Values represent mean ± SEM; *p<0.05 versus DSS+methocel.

Mice were exposed to 3.5% DSS in drinking water for eleven days and were treated with roflumilast (either 1 or 5 mg/kg/d orally once daily for 11 days, n = 8) or 4% methocel (n = 5). At day 11 the colon was removed, weighed, vortexed in PBS and centrifuged. TNFα was quantified in the eluate by ELISA. Values represent mean ± SEM; *p<0.05.