Human Nutrition & Food Science
Non Technical Summary
Patchouli essential oils have been used for medicinal applications due to a variety of biological effects including antibacterial, antifungal, antiinfluenza and antioxidant activities. Patchouli is a species of plant from the genus Pogostemon and major component of patchouli oil. Recently we found that patchouli alcohol possesses anti-inflammatory activity in immune cells. The general aim of this proposal is to study if patchouli alcohol influences occurrence of metabolic disorders including inflammation, obesity and cancer, focusing on elucidating the mechanisms and biological targets. The expected results will expand our understanding on health benefit of using patchouli alcohol and the scope of action using essential oil prevention of human chronic diseases.
Animal Health Component
Research Effort Categories
Goals / Objectives
Obesity is a worldwide epidemic associated with increased risk of various types of cancer. Chronic inflammation is a central characteristic of obesity, leading to many complications such as cancer. In particular, obesity-induced inflammation confers additional cancer risk beyond obesity itself.Patchouli alcohol is a tricyclic sesquiterpene and most abundant form in patchouli oil. Recently, we reported that patchouli alcohol possesses anti-inflammatory activity in mouse macrophages and colon cells (1). However, health benefits effects and reliable mechanisms by patchouli alcohol on metabolic disorders and human chronic diseases remain unanswered. The primary goal of this proposal is to explore if patchouli alcohol prevents inflammation, obesity and cancer. The second goal is to determine proposed mechanisms and molecular targets by which patchouli alcohol reduces inflammatory response in macrophages, differentiation of preadipocytes and proliferation of colonocytes.In the following specific aims we will investigate:Aim 1. Determine anti-inflammatory activity of patchouli alcohol Aim 2. Determine anti-obese and anti-diabetic activity of patchouli alcoholAim 3. Determine anti-cancer activity of patchouli alcohol
Aim 1. Determine anti-inflammatory activity of patchouli alcoholAim 1.1. In vivo study to exam anti-inflammatory activity of patchouli alcohol using DSS-induced colitis modelExperimental designThirty six experimental mice (C57BL/6 background) will be assigned to three groups and each will be receiving 1) vehicle (0.5% methylcellulose), 2) low dose of patchouli alcohol (20 mg/kg body weight) and 3) high dose of patchouli alcohol (40 mg/kg body weight) (twelve mice/each group). Patchouli alcohol suspension will be given by oral gavage every day for 2 weeks before DSS treatment. Then 5% (w/v) of DSS will be given to mice through autoclaved drinking water for one week with gavage of patchouli alcohol. Aftertreatment of DSS and patchouli alcohol, the mice will be euthanized.MeasurementsExtent of colitis, immunohistochemistry, MPO activity and gene expressionAim 1.2. In vitro studies to exam the mechanism(s) of anti-inflammatory activity using THP-1 monocytesExperimental designHuman monocytic cell line (THP-1) will be pretreated with different concentrations of patchouli alcohol (0, 12, 25, 50, 100 µM) and then co-treated with LPS or IL-6 for 30 minutes or 24hours.MeasurementsTranscriptional activity of NF-?B, nuclear localization of p65 and STAT3, proteasomal degradation of p65, phosphorylation of JNK/p38 MAPK, JAK1/STAT3 and ERK, and expression of target genesAim 2. Determine anti-obese and anti-diabetic activity of patchouli alcoholAim 2.1. In vivo study to exam anti-obese and anti-diabetic activity of patchouli alcohol using Ay/J mouse modelExperimental designAy/J mice in a B6 genetic background (KK.Cg-Ay/J, Jackson Laboratories) will be obtained and bred to produce enough numbers for experiment. At 6 weeks old, thirty six Ay/J mice will be assigned to three groups and each will be exposed to 1) vehicle (0.5% methylcellulose), 2) low dose of patchouli alcohol (20 mg/kg body weight), and 3) high dose of patchouli alcohol (40 mg/kg body weight) respectively (twelve mice/each group) for 4 weeks.MeasurementsBody fat pad, lipolysis/fatty acid oxidation, glucose tolerance test, macrophage infiltration, histological analysis and expression of chemokines and inflammatory cytokinesAim 2.2. In vitro studies to exam the mechanism(s) of anti-obese and anti-diabetic activity using 3T3-L1 preadipocytes and C2C12 myocytesExperimental design3T3-L1 preadipocytes will be cultured in DMEM containing 10% FBS until confluent. Two days after post-confluence, cells will be stimulated to differentiate with 1 μM dexamethasone, 0.5 mM isobutylmethylxanthine, and 5 μg/mL insulin in 10% FBS/DMEM for 2 days (day 0). Cells will be then maintained in 10% FBS/DMEM medium with 5 μg/mL of insulin for another 2 days, followed by culturing with 10% FBS/DMEM medium until analysis. Patchouli alcohol (0, 12, 25, 50, 100 µM) will be treated from day 0 to day 4 (for study of gene expression) or day 8 (for triglyceride accumulation). For culture of myocytes, C2C12 cells will be maintained in DMEM supplemented with 10% FBS and then differentiated using DMEM replacing 1% FBS in the absence or presence of different doses of patchouli alcohol (0, 12, 25, 50, 100 µM) for 7 days.MeasurementsLipid accumulation, in vitro glucose uptake and lipolysis, expression of adipogenesis and lipogenesis marker genes, phosphorylation of AMPKα and ACC, expression and translocation of β-catenin protein, and luciferase activity of TOP/FOP flashAim 3. Determine anti-cancer activity of patchouli alcoholAim 3.1. In vivo study to exam anti-cancer activity of patchouli alcohol using inflammation-induced colon cancer model Experimental designThirty six ApcMin/+ mice at 6 weeks of age will be assigned to three groups and each will be exposed to 1) vehicle (0.5% methylcellulose), 2) low dose of patchouli alcohol (20 mg/kg body weight), and 3) high dose of patchouli alcohol (40 mg/kg body weight) respectively (twelve mice/each group) by oral gavage for 4 weeks. Then 2% (w/v) DSS will be given to mice through autoclaved drinking water for one week and the patchouli alcohol will be gavaged every day for 4 weeks.Measurements Tumorigenesis in colon and histological analysisAim 3.2. In vitro studies to exam the mechanism(s) of anti-cancer activity using SW480 colonocytesExperimental designSW480 cells will be treated with different concentrations (0, 12, 25, 50, 100 µM) of patchouli alcohol for 0, 24 and 48 hours.MeasurementsApoptosis, activity of caspases and expression of apoptosis-associated genes, cell cycle distribution and expression of growth arrest genes, cell invasion/migration and expression of MMP-2/9, In vitro angiogenesis and expression of VEGF