Investigating Effects of Atorvastatin and Lactulose on ameliorating Crohn’s Disease in 2,4,6-Trinitrobenzene Sulphonic Acid (TNBS)-Induced Colitis: A Research Protocol

Introduction: Crohn’s disease (CD) is an inflammatory bowel disease (IBD) triggered by excessive activation of the gastrointestinal (GI) immune system, causing intestinal inflammation. Current therapeutic strategies use probiotics and fecal matter transplantation (FMT) to increase microbiome diversity and induce CD remission, but their efficacy is unclear. Conversely, prebiotics such as lactulose were found to increase anti-inflammatory bacteria thus reducing inflammatory markers in colitis mouse models. Additionally, atorvastatin has been found to modulate GI immune system activity by reducing chemokine expression, providing a potential treatment for CD. In this research protocol, a combination therapy using lactulose and atorvastatin is proposed as a treatment strategy for CD. Methods: 2.5% 2,4,6-trinitrobenzene sulphonic acid (TNBS) was injected intrarectally at a dose of 100 mg/kg to induce colitis in the mouse models. In this randomized controlled trial, the mice will be equally assigned to 5 groups: untreated TNBS mice (n = 10); TNBS mice treated with lactulose (0.005 ml / g of body weight, n = 10); TNBS mice treated with atorvastatin (5 mg/kg, n = 10); TNBS mice treated with the atorvastatin and lactulose combination therapy (n = 10); and the untreated healthy mouse (n = 10) to serve as the negative control. Histopathological analysis was conducted on colon tissue along with ELISA identification of inflammatory markers and mRNA analysis. Anticipated Results: It is expected that the TNBS group treated with the combination therapy will have the lowest levels of inflammatory cytokines. It is also expected that the treated TNBS mice will have the closest Crohn’s Disease Activity Index (CDAI) score to the healthy untreated condition. Discussion: Findings indicate that atorvastatin and lactulose combined supplementation could provide a more optimized treatment strategy for CD as opposed to atorvastatin or lactulose alone. Conclusion: The protocol anticipates the combined treatment to significantly improve colonic health in TNBS-induced colitis mice, resulting in decreased colonic ulcerations, diarrhea, and bowel wall thickness. Future studies should explore other inflammatory markers like CRPs and include different animal models to gain insights into treatment dosage and efficacy.