Toyoshi Umezu

Ambulation-promoting effect of peppermint oil and identification of its active constituents

Various plant-derived essential oils (EOs) have traditionally been used in the treatment of mental disorders, despite a lack of scientific evidence. In a previous study, we demonstrated that certain EOs possess behavioral effects, a finding that supports our original hypotheses that EOs possess psychoactive actions. The present study was conducted in order to obtain further evidence to support our hypothesis. Peppermint oil, a type of EO, is believed to be effective for treating mental fatigue. When the oil was administered intraperitoneally to ICR mice, the ambulatory activity of mice increased dramatically. We identified alpha-pinene, beta-pinene, (R)-(+)-limonene, 1,8-cineol, isomenthone, menthone, menthol, (R)-(+)-pulegone, menthyl acetate and caryophyllene as constituent elements of peppermint oil by GC-MS analysis. We then examined the effect of each constituent element of peppermint oil on ambulatory activity in mice. Intraperitoneal administration of 1,8-cineol, menthone, isomenthone, menthol, (R)-(+)-pulegone, menthyl acetate and caryophyllene significantly increased ambulatory activity in mice, suggesting that these chemicals are the behaviorally active elements of peppermint oil. Intravenous administration of these substances to mice induced a significant increase in ambulatory activity at much lower doses. The present study provides further evidence demonstrating that EOs possess pharmacological actions on behavior. In addition, our finding revealed that the action of peppermint oil comes from its constituent elements.

Anticonflict effects of lavender oil and identification of its active constituents

The pharmacological effects of lavender oil were investigated using two conflict tests in ICR mice, and then the active constituents were identified. Lavender oil produced significant anticonflict effects at 800 and 1600 mg/kg in the Geller conflict test and at 800 mg/kg in the Vogel conflict test, suggesting that the oil has an anti-anxiety effect. Analysis using GC/MS revealed that lavender oil contains 26 constituents, among which alpha-pinene (ratio, 0.22%), camphene (0.06%), beta-myrcene (5.33%), p-cymene (0.3%), limonene (1.06%), cineol (0.51%), linalool (26.12%), borneol (1.21%), terpinene-4-ol (4.64%), linalyl acetate (26.32%), geranyl acetate (2.14%) and caryophyllene (7.55%) were identified. We examined the effects of linalool, linalyl acetate, borneol, camphene, cineol, terpinen-4-ol, alpha-pinene and beta-myrcene using the Geller and Vogel conflict tests in ICR mice. Cineol, terpinen-4-ol, alpha-pinene and beta-myrcene did not produce any significant anticonflict effects in the Geller test. Linalyl acetate did not produce any significant anticonflict effects in either test. Both borneol and camphene at 800 mg/kg produced significant anticonflict effects in the Geller, but not in the Vogel conflict test. Linalool, a major constituent of lavender oil, produced significant anticonflict effects at 600 and 400 mg/kg in the Geller and Vogel tests, respectively, findings that were similar to those of lavender oil. Thus, we concluded that linalool is the major pharmacologically active constituent involved in the anti-anxiety effect of lavender oil.

Anticonflict effects of rose oil and identification of its active constituents

The present study investigates the pharmacologically active constituents of rose oil, which possesses anti-conflict effects. Analysis using GC/MS revealed that rose oil contains 9 substances that were identified as myrcene, benzyl alcohol, 2-phenethyl alcohol, citronellol, geraniol, citronellyl acetate, eugenol, geranyl acetate and methyl eugenol. We examined the effects of each of these substances using the Geller and Vogel conflict tests in ICR mice. Myrcene, benzyl alcohol and citronellyl acetate did not produce any effects in either tests. Geranyl acetate and methyl eugenol produced no effect in the Geller conflict test. Geraniol and eugenol decreased the response rate during the safe period of the Geller conflict test, but did not affect the response rate during the alarm period. In contrast, 2-phenethyl alcohol and citronellol, like rose oil, produced an increasing effect on the response rate during the alarm period in the Geller conflict test. In addition, both chemicals increased the number of electric shocks mice received in the Vogel conflict test in a manner similar to that of rose oil. Given that 2-phenethyl alcohol and citronellol produced the same anti-conflict effects in both tests as rose oil, we concluded that they are the pharmacologically active constituents of anti-anxiety-like effect of rose oil.

Anticonflict effects of plant-derived essential oils.

The present study examined the pharmacological actions of four different plant-derived essential oils (rose, ylang-ylang, camomile, orange) in two types of conflict tests using ICR mice. In the Vogel conflict test, in which any drinking behavior of the mice was punished by an electric shock, the benzodiazepine agonist, diazepam (DZ), increased the number of electric shocks the mice received. This number increased after administration of rose oil. In contrast, ylang-ylang, camomile, and orange oil did not produce such an effect in this test. In the Geller conflict test where lever-pressing of mice was reinforced by food pellets and then punished by electric shock, response (lever-pressing) rate during the alarm period was increased as well by the positive control drug, DZ. Similarly, the response rate during the alarm period increased after administration of rose oil. Here as well, ylang-ylang, camomile, and orange oils did not produce an anticonflict effect. In the Vogel conflict test, the anticonflict effect of DZ was reversed by the benzodiazepine antagonist, flumazenil (Ro15-1788) (FL). However, the effect of rose oil in this test was not antagonized by FL. The present study showed that rose oil possesses anticonflict effects, and that the effects are not mediated by the benzodiazepine binding site of the GABA(A) receptor complex. Such pharmacological actions may at least partially account for human behavioral effects attributed to essential oils.

Evidence for dopamine involvement in ambulation promoted by pulegone in mice

The present study examines the mechanism that underlies the ability of menthone (MTN), a constituent of peppermint oil, to promote mouse ambulation. Since bupropion (BUP), a dopamine (DA) uptake inhibitor, promotes mouse ambulation, the effect of MTN combined with BUP on ambulation was investigated. The results showed that BUP with MTN produced an additive interaction on mouse ambulation. The effects of DA antagonists chlorpromazine, fluphenazine, haloperidol, SCH12679 and spiperone on the ability of MTN to promote ambulation were then examined. All of these antagonists attenuated the effects of MTN. Prior exposure to the tyrosine hydroxylase inhibitor alpha-methyl-p-tyrosine, which inhibits catecholamines synthesis, decreased subsequent sensitivity to the effect of MTN. These results suggest that DA is involved in the ability of MTN to promote ambulation in mice.

Behavioral effects of trichloroethylene and tetrachloroethylene in mice.

This study was performed to clarify the toxicological profiles of trichloroethylene (TRCE) and tetrachloroethylene (TECE) when they are administered intraperitoneally in mice. The ED50 for loss of righting reflex were 2596 mg/kg in TRCE and 4209 mg/kg in TECE. TRCE and TECE impaired bridge test performance at 500 and 2000 mg/kg, respectively. An operant behavior performance was also inhibited by TRCE at 1000 mg/kg and by TECE at 2000 mg/kg. Both TRCE and TECE exhibited anticonflict effects in a Vogel-type task at 500 mg/kg. This effect was confirmed by the finding that TRCE exhibited anticonflict action in a Geller-type paradigm at 250 mg/kg and more, as did TRCE did at 1000 mg/kg. These results show that TRCE and TECE affect various behaviors in mice and suggest that conflict behaviors are one of the most sensitive behavioral indicators of the effects of these substances. The toxicological profiles of TRCE and TECE with respect to behavioral effects were very similar, and they can be classified in a single category.

Diphenylarsinic acid produces behavioral effects in mice relevant to symptoms observed in citizens who ingested polluted well water.

Citizens in an area of Kamisu City, Ibaraki, Japan had exhibited unusual health problems, and pollution of well water by diphenylarsinic acid (DPAA) was found in the area. We examined the effects of DPAA on various behaviors in mice. DPAA was administered to mice through free intake of drinking water for 27 weeks (subchronic exposure) or 57 weeks (chronic exposure), and behavior was examined during exposure. DPAA at 30-100 ppm increased ambulatory activity and the response rate of the shuttle type discrete conditioned avoidance response of mice. DPAA reduced coordination ability on the fixed rod at 100 ppm. DPAA at 7.5-15 ppm also reduced coordination on the rotating rod, although these doses of DPAA did not affect coordination on the fixed rod. Chronic exposure to 7.5-15 ppm of DPAA produced anti-anxiety-like effects in the elevated plus maze test, whereas subchronic exposure to 100 ppm of DPAA produced anxiogenic-like effects. Neither subchronic nor chronic exposure to 7.5-100 ppm of DPAA affected learning ability and/or memory, as evaluated using the passive avoidance response. Exposure to 15-30 ppm of DPAA for 52 weeks did not alter weights of the cerebrum and cerebellum or amounts of neuron marker protein TUJ-1 or astrocyte marker protein glial fibrillary acidic protein in the cerebellum of mice. Behavioral effects observed in mice seem relevant to symptoms observed in patients from Kamisu City.

Evaluation of the Effects of Plant‐derived Essential Oils on Central Nervous System Function Using Discrete Shuttle‐type Conditioned Avoidance Response in Mice

Although plant‐derived essential oils (EOs) have been used to treat various mental disorders, their central nervous system (CNS) acting effects have not been clarified. The present study compared the effects of 20 kinds of EOs with the effects of already‐known CNS acting drugs to examine whether the EOs exhibited CNS stimulant‐like effects, CNS depressant‐like effects, or neither. All agents were tested using a discrete shuttle‐type conditioned avoidance task in mice. Essential oils of peppermint and chamomile exhibited CNS stimulant‐like effects; that is, they increased the response rate (number of shuttlings/min) of the avoidance response. Linden also increased the response rate, however, the effect was not dose‐dependent. In contrast, EOs of orange, grapefruit, and cypress exhibited CNS depressant‐like effects; that is, they decreased the response rate of the avoidance response. Essential oils of eucalyptus and rose decreased the avoidance rate (number of avoidance responses/number of avoidance trials) without affecting the response rate, indicating that they may exhibit some CNS acting effects. Essential oils of 12 other plants, including juniper, patchouli, geranium, jasmine, clary sage, neroli, lavender, lemon, ylang‐ylang, niaouli, vetivert and frankincense had no effect on the avoidance response in mice. Copyright

Unusual Effects of Nicotine as a Psychostimulant on Ambulatory Activity in Mice

The present study examined the effect of nicotine, alone and in combination with various drugs that act on the CNS, on ambulatory activity, a behavioral index for locomotion, in ICR (CD-1) strain mice. Nicotine at 0.25–2 mg/kg acutely reduced ambulatory activity of ICR mice. The effect of nicotine was similar to that of haloperidol and fluphenazine but distinct from that of bupropion and methylphenidate. ICR mice developed tolerance against the inhibitory effect of nicotine on ambulatory activity when nicotine was repeatedly administered. This effect was also distinct from bupropion and methylphenidate as they produced augmentation of their ambulation-stimulating effects in ICR mice. Nicotine reduced the ambulation-stimulating effects of bupropion and methylphenidate as well as haloperidol and fluphenazine. Taken together, nicotine exhibited unusual effects as a psychostimulant on ambulatory activity in ICR mice.

Different behavioral effect dose-response profiles in mice exposed to two-carbon chlorinated hydrocarbons: influence of structural and physical properties.

The present study aimed to clarify whether dose-response profiles of acute behavioral effects of 1,2-dichloroethane (DCE), 1,1,1-trichloroethane (TCE), trichloroethylene (TRIC), and tetrachloroethylene (PERC) differ. A test battery involving 6 behavioral endpoints was applied to evaluate the effects of DCE, TCE, TRIC, and PERC in male ICR strain mice under the same experimental conditions. The behavioral effect dose-response profiles of these compounds differed. Regression analysis was used to evaluate the relationship between the dose-response profiles and structural and physical properties of the compounds. Dose-response profile differences correlated significantly with differences in specific structural and physical properties. These results suggest that differences in specific structural and physical properties of DCE, TCE, TRIC, and PERC are responsible for differences in behavioral effects that lead to a variety of dose-response profiles.