LAVENDER PROTECTS AGAINST INFECTIOUS COLITIS

JESSICA BAKERSUPERVISOR: DR. DEANNA GIBSONUBC 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 mouse-specific 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

Group 1: Mineral oil OR

Group 2: Lavender oil

Orally gavaged throughout 10-day infection

B6 (C57BL/6)Mice

OR

C3H (C3H/HeOuJ)Mice

Orally gavaged with C. rodentium for 10 days

Monitored morbidity and mortality

PROCEDURE:

Lower Gastrointestinal Tract

Cecum

Distal Colon

RNAlater: extract the RNA for q-PCR

H&E stain: Scoring

Mesenteric Lymph Nodes

AND

Spleen

Bacterial counts toassess systemic infection

• 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)

B6 Mice

C3H Mice

MICROGRAPHS OF THE CECUM

Lavender Oil significantly reduces• Edema• Infiltration • Epithelial hyperplasia • Goblet cell depletion• Epithelial damage (P< 0.01, Mann-Whitney T-test)

Uninfected Infected + Mineral Oil Infected + Lavender Oil

IN VITRO AND IN VIVO EVIDENCE THAT LAVENDER OIL CAN DIRECTLY KILL THIS PATHOGEN

Mineral Oil

Lavender Oil

Lavender Mineral Oil Oil

Colo

ny F

orm

ing

Uni

ts

MLN

Spleen

In Vivo In Vitro

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

Two mechanismsAntimicrobial Immune Modulation

Terpene Okanagan Lavender Oil

Cineole (%) 27.09 ± 1.78Camphor (%) 3.03 ± 0.26Linalool (%) 36.28 ± 4.16Linalool acetate (%) 1.86 ± 0.36Borneol (%) 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