Crohn`s disease is a severe chronic inflammation that is treated mainly by immunosuppression, which often has serious side effects. There is a need to develop new drugs for treating this disease that have few side effects. Heme oxygenase-1 (HO-1) has immunosuppressive properties, but the mechanism of its anti-inflammatory actions is unclear. We investigated the protective effects of HO-1 on trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. An HO-1 inducer, cobalt protoporphyrin IX (CoPPIX), dramatically improved the clinical and histopathological symptoms in TNBS-induced colitis. CoPPIX suppressed tumor necrosis factor-alpha and interleukin-1beta expression and down-regulated the nuclear transcription factor kappa B activity caused by TNBS. The vehicle copper protoporphyrin IX (CuPPIX) failed to duplicate the protective effects seen with CoPPIX. Moreover, an inhibitor of HO-1 activity-zinc protoporphyrin IX-reversed the protective effects of CoPPIX on TNBS-induced colitis. In conclusion CoPPIX protects against TNBS-induced colonic damage by inducing HO-1, which might be an important target in the treatment of Crohn`s disease.
Animals ; Cobalt ; Colitis* ; Colon ; Copper ; Heme Oxygenase-1 ; Immunosuppression ; Inflammation ; Interleukin-1beta ; Mice ; Transcription Factors ; Tumor Necrosis Factor-alpha

BACKGROUND: Generation of perfluorocarbon-exposed sonicated dextrose albumin (PESDA), the custom-made contrast agent, is performed under certain conditions that have been proposed by its original developer. We doubted whether the known composition and manufacturing method of PESDA is ideal and if there is an optimal method of storing batches of PESDA for a significant time duration. METHODS: PESDA was generated with several different composition of ingredients (5% human serum albumin, 5% dextrose water, and perfluorocarbon (PFC) gas), where various ratios of each were used. Sonication was performed for various durations. After manufacturing, the mean size and concentration of the microbubbles were evaluated by hemocytometer and compared. The generated PESDA was stored for 48 hours under 4 degrees C or -20 degrees C and changes in size and concentration of microbubbles were evaluated and compared. RESULTS: The best concentration of microbubbles was found with a mix ratio of albumin: PFC: dextrose of 1:1:1 and sonication time of 90 sec. The microbubble concentration of the optimal PESDA was not different to that of the conventionally manufactured one (9.47+/-1.70 x 10(8) /mL vs. 8.34+/-0.87 x 10(8) /mL, p>0.05) but the mean microbubble size was significantly smaller (1.22+/-0.31 um vs. 1.66+/-0.32 um, p<0.01). After 48 hours, the concentration of microbubbles was reduced by 34+/-3% (p=NS) and 55+/-0.2% (p<0.05) and the size increased by 77+/-25% and 108+/-41% (p=NS in both) in the 4 degrees C -stored and -20 degrees C -stored PESDA, respectively. CONCLUSION: The optimal composition of PESDA ingredients is 1:1:1 for albumin, PFC, and dextrose water, and the best duration of sonication is 90 seconds. Refrigeration under 4 degrees C may be the best way for storage of PESDA for 48 hours.
Echocardiography ; Glucose* ; Humans ; Microbubbles ; Refrigeration ; Serum Albumin ; Sonication ; Water