title: Experimental evidence of heparanase, Hsp70 and NF-κB gene expression on the response of anti-inflammatory drugs in TNBS-induced colonic inflammation.
abstract: OBJECTIVE
  Etiopathogenesis of inflammatory bowel disease is unclear and results from a complex interplay of genetic, microbial, environmental and immune factors. Elucidating the mechanisms that drive IBD depends on the detailed characterization of human inflammatory mediators in animal models. Therefore, we studied how intestinal inflammation affects heparanase, NF-κB and Hsp70 gene expression in rats, and if current intestinal anti-inflammatory drugs (sulphasalazine, prednisolone and azathioprine) act on these expressions. Moreover, we investigated the relationships among these genes with colonic cytokines levels (IL-1β, TNF-α, IL-6, INF-γ and IL-10) and oxidative stress that have fundamental role in IBD.
  
  
  METHODS
  Macroscopic parameters (diarrhea, extension of lesion, colonic weight/length ratio and damage score), biochemical markers (myeloperoxidase and alkaline phosphatase activities, and glutathione, IL-1β, TNF-α, IL-6, INF-γ and IL-10 levels), gene expressions (heparanase, NF-κB and Hsp70), and microscopic evaluations (optic, electronic scanning and transmission microscopic) were performed in rats.
  
  
  RESULTS
  Expression of heparanase, Hsp70 and NF-κB and oxidative stress were increased by inflammatory process and differentially modulated by sulphasalazine, prednisolone and azathioprine treatments. Protective effects of drugs were also related to differential modulation of cytokine changes induced by inflammatory process, showing different mechanisms to control inflammation.
  
  
  CONCLUSIONS
  Heparanase, NF-κB and Hsp70 gene expression participate in the inflammatory response induced by TNBS and represent pharmacological targets of the intestinal anti-inflammatory drugs. In addition, current drugs used to treat IBD (sulphasalazine, prednisolone and azathioprine) differentially modulate heparanase, NF-κB and Hsp70 gene expression, cytokine production and oxidative stress.
authors.affiliation: Laboratory of Phytomedicines, Pharmacology and Biotechnology (PhytoPharmaTech), Department of Pharmacology, Institute of Biosciences, São Paulo State University - UNESP, Botucatu, São Paulo CEP 18618-000, Brazil.
authors.forename: Ana E V
authors.initials: AE
authors.lastname: Quaglio
authors.affiliation: Laboratory of Phytomedicines, Pharmacology and Biotechnology (PhytoPharmaTech), Department of Pharmacology, Institute of Biosciences, São Paulo State University - UNESP, Botucatu, São Paulo CEP 18618-000, Brazil.
authors.forename: Anthony C S
authors.initials: AC
authors.lastname: Castilho
authors.affiliation: Laboratory of Phytomedicines, Pharmacology and Biotechnology (PhytoPharmaTech), Department of Pharmacology, Institute of Biosciences, São Paulo State University - UNESP, Botucatu, São Paulo CEP 18618-000, Brazil. Electronic address: ldistasi@ibb.unesp.br.
authors.forename: Luiz C
authors.initials: LC
authors.lastname: Di Stasi
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001379:Azathioprine / Q000494:pharmacology; D015415:Biomarkers / Q000032:analysis; D003092:Colitis / Q000139:chemically induced* / Q000188:drug therapy* / Q000235:genetics; D003106:Colon / Q000378:metabolism / Q000473:pathology; D016207:Cytokines / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D005966:Glucuronidase / Q000096:biosynthesis* / Q000235:genetics*; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics*; D015212:Inflammatory Bowel Diseases / Q000235:genetics / Q000378:metabolism; D008297:Male; D016328:NF-kappa B / Q000096:biosynthesis* / Q000235:genetics*; D011239:Prednisolone / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012460:Sulfasalazine / Q000494:pharmacology; D014302:Trinitrobenzenesulfonic Acid; D000099175:Heparanase
chemical_list: D000893:Anti-Inflammatory Agents; D001379:Azathioprine; D015415:Biomarkers; D016207:Cytokines; D005966:Glucuronidase; D018840:HSP70 Heat-Shock Proteins; D016328:NF-kappa B; D011239:Prednisolone; D012460:Sulfasalazine; D014302:Trinitrobenzenesulfonic Acid; D000099175:Heparanase
keywords: Heparanase; Hsp70; Inflammatory bowel disease; Intestinal anti-inflammatory drugs; Intestinal inflammation; NF-κB