LAVENDER PROTECTS AGAINST INFECTIOUS COLITIS JESSICA BAKER SUPERVISOR: DR. DEANNA GIBSON UBC MURC 2011 INFLAMMATORY BOWL DISEASE (IBD): COLITIS IBD: Ulcerative Colitis and Crohn’s Disease It is a chronic, relapsing, immunologically-mediated disorder in the gastrointestinal tract Unknown cause, although onset and activation is thought to develop in predisposed individuals Currently there are limited therapeutic options BACTERIA-INDUCED COLITIS MODEL Use a bacterium called Citrobacter rodentium: a mousespecific intestinal pathogen Some important inflammatory factors: TNF-α, IFN-γ, iNOS, IL-22, MIP-2α and IL-10 Results in intestinal damage similar to human colitis LAVANDULA X INTERMEDIA: OKANAGAN LAVENDER Developed in Dr. Soheil Mahmoud’s Lab at UBC Okanagan Objective: to improve oil composition and yield Terpene Wild Type Oil Okanagan Lavender Oil Cineole (%) 13.73 ± 1.28 27.09 ± 1.78 Camphor (%) 11.16 ± 0.35 3.03 ± 0.26 Linalool (%) 35.92 ± 2.27 36.28 ± 4.16 Linalool acetate (%) 25.34 ± 2.34 1.86 ± 0.36 Borneol (%) 3.55 ± 0.59 21.01 ± 3.63 Desautels, A, et al. “Suppression of Linalool Acetate production in Lavandula x intermedia.”Natural Products Communications 4.11 (2009): 1533-1536. Santos, F.A., et al. “1,8-cineole (eucalyptol), a monoterpene oxide attenuates the colonic damage in rats on acute TNBS-colitis.” Food and Chemical Toxicology 42 (2004): 579-584. Tung, Yu-Tang, et al. “Anti-inflammation activities of essential oil and its constituents from indigenous cinnamon (Cinnamomum osmophloeum) twigs.” Bioresource Technology 99 (2008): 3908-3913. HYPOTHESIS We hypothesize that Okanagan Lavender oil will decrease the severity of infectious colitis due to its ability to increase pathogen clearance as well as its ability to decrease inflammation and associated inflammatory mediators PROCEDURE: B6 (C57BL/6) Mice OR C3H (C3H/HeOuJ) Mice Group 1: Mineral oil OR Group 2: Lavender oil Orally gavaged throughout 10-day infection Orally gavaged with C. rodentium for 10 days Monitored morbidity and mortality Lower Gastrointestinal Tract Cecum RNAlater: extract the RNA for q-PCR Distal Colon H&E stain: Scoring Spleen AND Mesenteric Lymph Nodes Bacterial counts to assess systemic infection B6 Mice C3H Mice • Lavender-treated mice susceptible to lethal infectious colitis (C3H mice) showed a drastic decrease in mortality • Lavender-treated mice that survive colitis (B6 mice) showed significantly less morbidity (P< 0.05; Mann-Whitney T-test) MICROGRAPHS OF THE CECUM Uninfected Infected + Mineral Oil Lavender Oil significantly reduces • Edema • Infiltration • Epithelial hyperplasia • Goblet cell depletion • Epithelial damage (P< 0.01, Mann-Whitney T-test) Infected + Lavender Oil IN VITRO AND IN VIVO EVIDENCE THAT LAVENDER OIL CAN DIRECTLY KILL THIS PATHOGEN In Vivo In Vitro Colony Forming Units MLN Mineral Oil Lavender Oil Spleen Lavender Oil Mineral Oil Q-PCR RESULTS The mice orally gavaged with lavender oil showed a significant modulation in the immune response (P< 0.05, Mann-Whitney T-test) IMMUNOFLUORESCENCE RESULTS Infected + Infected + Lavender-treated mice showed a drastic decrease in immune cell infiltration CONCLUSION The significant decrease in morbidity, cecal tissue damage, pro-inflammatory cytokine production and immune cell infiltration provide strong support for our hypothesis that Okanagan Lavender Oil protects against infectious colitis Terpene Two mechanisms Antimicrobial Immune Modulation Okanagan Lavender Oil Cineole (%) 27.09 ± 1.78 Camphor (%) 3.03 ± 0.26 Linalool (%) 36.28 ± 4.16 Linalool acetate (%) 1.86 ± 0.36 Borneol (%) 21.01 ± 3.63 FUTURE Results so far warrant further analysis Isolate and test Lavender components Need to do more animal studies with higher N number and varied dose amounts Need to examine overall health in other areas of the body Potential outcomes: An anti-inflammatory drug for chronic intestinal diseases such as colitis An antimicrobial drug for intestinal infectious diseases ACKNOWLEDGMENTS Dr. Deanna Gibson (Supervisor) Dr. Soheil Mahmoud and Lab Dr. Sanjoy Ghosh Samantha Makarenko Kirsty Brown Dr. Mark Rheault Brie Matier Irving K. Barber Endowment fund Everyone with MURC 2011