The application and relevance of ex vivo culture systems for assessment of IBD treatment in murine models of colitis.
The aim of this study was to investigate the relevance of mouse ex vivo cultures as a first screening model for new therapeutic agents of Inflammatory Bowel Disease (IBD). Two murine models (dextran sodium sulphate (DSS)-induced colitis and Galphai2-deficient mice) and two anti-inflammatory agents (methyl-prednisolone and the proteasome inhibitor MG132) were evaluated. The in vivo effects of methyl-prednisolone were assessed in both models. Ex vivo colonic tissue from both mouse models were cultured in the presence or absence of the drugs and TaqMan Low-Density arrays were used to assess the regulation of inflammatory genes before and after drug treatment. Colitis induced a similar inflammatory gene profile in both mouse models in in vivo studies and in ex vivo cultures. The differences encountered reflected the different phases of colitis in the models, e.g. innate cytokine/chemokine profile in the DSS model and T cell related markers in Galphai2-deficient mice. After steroid treatment, a similar pattern of genes was suppressed in the two mouse models. We confirmed the suppression of inflammatory gene expression for IL-1beta, IL-6 and iNOS in ex vivo and in vivo colons from both mouse models by quantitative RT-PCR. Importantly, the inflammatory responses in the murine ex vivo culture system reflected the in vivo response in the inflamed colonic tissue as assessed by changes in inflammatory gene expression, suggesting that the murine culture system can be used for validation of future IBD therapies.
Fritsch Fredin
Maria
M
Department of Bioscience at AstraZeneca R&D Mölndal, Sweden. maria.fritsch@astrazeneca.com
Vidal
Alexander
A
Utkovic
Helena
H
Götlind
Yu-Yuan
YY
Willén
Roger
R
Jansson
Liselotte
L
Hultgren Hörnquist
Elisabeth
E
Melgar
Silvia
S
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000627:therapeutic use; D002478:Cells, Cultured; D003092:Colitis / Q000139:chemically induced / Q000188:drug therapy* / Q000235:genetics; D017931:DNA Primers; D016264:Dextran Sulfate; D005260:Female; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000172:deficiency / Q000235:genetics; D015212:Inflammatory Bowel Diseases / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D053583:Interleukin-1beta / Q000096:biosynthesis; D015850:Interleukin-6 / Q000096:biosynthesis; D007976:Leupeptins / Q000494:pharmacology; D008775:Methylprednisolone / Q000627:therapeutic use; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis; D009924:Organ Culture Techniques; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000893:Anti-Inflammatory Agents; D017931:DNA Primers; D053583:Interleukin-1beta; D015850:Interleukin-6; D007976:Leupeptins; D016264:Dextran Sulfate; D052247:Nitric Oxide Synthase Type II; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde; D008775:Methylprednisolone