Tissue-specific enhancement of xenobiotic detoxification enzymes
in mice by dietary rosemary extract.
Singletary KW; Rokusek JT
Department of Food Science and Human Nutrition, University of
Illinois, Urbana, IL 61801, USA.
Plant Foods Hum Nutr, 50(1):47-53 1997
Plant foods contain nutritive and minor, nonnutritive components
capable of inhibiting experimental carcinogenesis. Many of these
cancer-protective extracts act by enhancing the activities of
enzymes that can detoxify reactive substances. In the present
study an extract of the spice plant rosemary was fed at concentrations
of 0.3% and 0.6% (by weight) for 4 weeks to female A/J mice prior
to determination of the activities of the detoxification enzymes
glutathione-S-transferase (GST) and NAD(P)H: quinone reductase
(QR) in lung, liver and stomach. Liver activities of GST and
QR, and stomach GST activity were significantly increased in
animals fed diets containing rosemary extract. However, diets
supplemented with rosemary extract did not affect lung GST and
QR activities. These results indicate that components of rosemary
extract have the potential to protect mouse liver and stomach
from carcinogenic or toxic agents.
rosemary extract and carnosol stimulate rat liver glutathione-S-transferase
and quinone reductase activities.
Department of Food Science and Human Nutrition, University of
Illinois, Urbana, 61801, USA.
Cancer Lett, 100(1-2):139-44 1996 Feb 27
The effects of dietary intake and intraperitoneal (i.p.) administration
of an extract of the spice rosemary and of the rosemary constituent
carnosol on the liver activities of glutathione-S-transferase
(GST) and NAD(P)H-quinone reductase (QR) in the female rat were
evaluated. rosemary extract at concentrations from 0.25 to 1.0%
(by wt.) in the diet resulted in a significant 3.5- to 4.5-fold
increase in liver GST and a 3.3- to 4.0-fold increase in liver
QR activities compared to controls. Carnosol supplemented in
the diet at levels from 0.01 to 1.0% did not enhance GST activity.
When rosemary extract and carnosol were administered i.p. there
was a significant increase in liver GST and QR activities. The
injection of rosemary extract (200 mg/kg) was associated with
1.5-fold and 3.2-fold increases in GST and QR activities, respectively,
compared to controls. The injection of carnosol at doses from
100 to 400 mg/kg was associated with 1.6- to 1.9-fold increases
in GST activity and 3.1- to 4.8-fold increases in QR activity,
compared to controls. These data indicate that rosemary extract
in the diet or injected i.p. and carnosol administered i.p. are
effective enhancers of the in vivo activity of liver GST and
QR in the female rat.
Inhibition of skin tumorigenesis by rosemary and its constituents
carnosol and ursolic acid.
Huang MT; Ho CT; Wang ZY; Ferraro T; Lou YR; Stauber K; Ma W;
Georgiadis C; Laskin JD; Conney AH
Department of Chemical Biology and Pharmacognosy, College of
Pharmacy, Rutgers, State University of New Jersey, Piscataway
Cancer Res, 54(3):701-8 1994 Feb 1
A methanol extract of the leaves of the plant Rosmarinus officinalis
L. (rosemary) was evaluated for its effects on tumor initiation
and promotion in mouse skin. Application of rosemary to mouse
skin inhibited the covalent binding of benzo(a)pyrene ÍB(a)PÍ
to epidermal DNA and inhibited tumor initiation by B(a)P and
7,12-dimethylbenzÍaÍanthracene (DMBA). Topical
application of 20 nmol B(a)P to the backs of mice once weekly
for 10 weeks, followed 1 week later by promotion with 15 nmol
12-O-tetradecanoylphorbol-13-acetate (TPA) twice weekly for 21
weeks, resulted in the formation of 7.1 tumors per mouse. In
a parallel group of animals that were treated topically with
1.2 or 3.6 mg of rosemary 5 min prior to each application of
B(a)P, the number of tumors per mouse was decreased by 54 or
64%, respectively. Application of rosemary to mouse skin also
inhibited TPA-induced ornithine decarboxylase activity, TPA-induced
inflammation, arachidonic acid-induced inflammation, TPA-induced
hyperplasia, and TPA-induced tumor promotion. Mice initiated
with 200 nmol DMBA and promoted with 5 nmol TPA twice weekly
for 19 weeks developed an average of 17.2 skin tumors per mouse.
Treatment of the DMBA-initiated mice with 0.4, 1.2, or 3.6 mg
of rosemary together with 5 nmol TPA twice weekly for 19 weeks
inhibited the number of TPA-induced skin tumors per mouse by
40, 68, or 99%, respectively. Topical application of carnosol
or ursolic acid isolated from rosemary inhibited TPA-induced
ear inflammation, ornithine decarboxylase activity, and tumor
promotion. Topical application of 1, 3, or 10 mumol carnosol
together with 5 nmol TPA twice weekly for 20 weeks to the backs
of mice previously initiated with DMBA inhibited the number of
skin tumors per mouse by 38, 63, or 78%, respectively. Topical
application of 0.1, 0.3, 1, or 2 mumol ursolic acid together
with 5 nmol TPA twice weekly for 20 weeks to DMBA-initiated mice
inhibited the number of tumors per mouse by 45-61%.
Inhibition of 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary
tumorigenesis and of in vivo formation of mammary DMBA-DNA adducts
by rosemary extract.
Singletary KW; Nelshoppen JM
Division of Foods and Nutrition, University of Illinois, Urbana-Champaign.
Cancer Lett, 60(2):169-75 1991 Nov
The effect of dietary intake of an extract of the spice plant
Rosmarinus officinalis L. on 7,12-dimethylbenz[a]anthracene (DMBA)-induced
mammary tumorigenesis and on the in vivo formation of mammary
DMBA-DNA adducts was evaluated. Supplementation of a semi-purified
diet with 1.0% (by wt.) rosemary extract resulted in a significant
(47%) decrease in mammary tumor incidence compared to controls.
In subsequent studies, dietary supplementation with 0.5% and
1.0% rosemary extract inhibited total in vivo binding of DMBA
to mammary epithelial cell DNA by an average of 42%. This decrease
in total binding was not due to a uniform decrease in the formation
of all mammary DMBA-DNA adducts. The formation of two major adducts
derived from the anti-diastereomer of DMBA and bound to deoxyguanosine
(anti-dGuo) was significantly decreased at both dietary rosemary
concentrations. The formation of the syn-dGuo adduct also was
inhibited, whereas formation of the syn-dAdo adduct was unaffected
by consumption of the rosemary extract. These studies suggest
that use of rosemary extract and its individual antioxidative
constituents as chemopreventative agents for experimental mammary
tumorigenesis warrant further investigation.
Induction of the anti-carcinogenic enzyme quinone reductase by
food extracts using murine hepatoma cells.
Tawfiq N; Wanigatunga S; Heaney RK; Musk SR; Williamson G; Fenwick
Department of Food Molecular Biochemistry, Norwich Laboratory,
Eur J Cancer Prev, 3(3):285-92 1994 May
Over 145 extracts of vegetables, fruits, herbs, spices and beverages
which are consumed regularly in the European diet have been surveyed
for potential anti-carcinogenic activity using an assay which
measures the induction of NAD(P)H: (quinone acceptor) menadione
oxidoreductase (quinone reductase, QR) activity in murine cells
challenged with solutions of potential inducers. When appropriate
the study has included extracts prepared from cooked and autolysed
material. The results indicate that extracts of some brassicas,
legumes (peas), lettuces, red pepper, grapefruit and some herbs
including basil, tarragon and rosemary are inducers of QR activity.
Inducing activity is strongly dependent on processing and on
Inhibition by rosemary and carnosol of 7,12-dimethylbenz[a]anthracene
(DMBA)-induced rat mammary tumorigenesis and in vivo DMBA-DNA
Singletary K; MacDonald C; Wallig M
Department of Food Science and Human Nutrition, University of
Urbana 61801, USA.
Cancer Lett, 104(1):43-8 1996 Jun 24
Extracts of the spice rosemary officinalis L. have been reported
to inhibit experimental carcinogenesis. Two rosemary components,
carnosol and ursolic acid, appear to be partly responsible for
the antitumorigenic activity of rosemary. The present studies
were conducted in order to evaluate the activity of rosemary
extract, carnosol and ursolic acid in inhibiting the in vivo
formation of mammary 7,12-dimethylbenz[a]anthracene (DMBA)-DNA
adducts and the initiation of DMBA-induced mammary tumorigenesis
in female rats. Supplementation of diets for 2 weeks with rosemary
extract (0.5% by wt) but not carnosol (1.0%) or ursolic acid
(0.5%) resulted in a significant decrease in the in vivo formation
of rat mammary DMBA-DNA adducts, compared to controls. When injected
intraperitoneally (i.p.) for 5 days at 200 mg/kg body wt, rosemary
and carnosol, but not ursolic acid, significantly inhibited mammary
adduct formation by 44% and 40%, respectively, compared to controls.
Injection of this dose of rosemary and carnosol was associated
with a significant 74% and 65% decrease, respectively, in the
number of DMBA-induced mammary adenocarcinomas per rat, compared
to controls. Ursolic acid injection had no effect on mammary
tumorigenesis. Therefore, carnosol is one rosemary constituent
that can prevent DMBA-induced DNA damage and tumor formation
in the rat mammary gland, and, thus, has potential for use as
a breast cancer chemopreventative agent.
Dietary rosemary suppresses 7,12-dimethylbenz(a)anthracene binding
to rat mammary cell DNA.
Amagase H; Sakamoto K; Segal ER; Milner JA
Department of Nutrition, The Pennsylvania State University, University
Park, 16802, USA.
J Nutr, 126(5):1475-80 1996 May
Commercially available ground rosemary powder was examined for
its ability to modify the in vivo binding of 7,12-dimethylbenz(a)anthracene
(DMBA) metabolites to mammary cell DNA in 55-d-old rats fed diets
containing varying quantities and types of lipids. Supplementing
a casein-based diet containing 20% corn oil with 1 % rosemary
for 2 wk reduced by 76% the occurrence of DMBA-induced DNA adducts
occurring 24 h after treatment with 50 mg DMBA/kg body weight.
A comparable reduction in DNA adducts (66%) occurred when 0.5%
rosemary was added to a diet containing 20% corn oil, and the
quantity of DMBA given was reduced to 25 mg/kg body weight. The
reduction in the occurrence of adducts occurring 24 h after DMBA
treatment caused by 0.5% dietary rosemary was greater (P <
0.05) when added to a diet containing 20% corn oil than when
added to a diet containing 5% corn oil and 15% coconut oil. rosemary,
1% but not 0.5%, reduced DMBA-induced DNA adducts when the diet
contained 5% corn oil. These studies demonstrate that rosemary
is effective in reducing the binding of DMBA metabolites to rat
mammary cell DNA. Furthermore, the present studies demonstrate
that the benefits of rosemary are dependent on the source and
concentration of dietary lipids.
rosemary components inhibit benzo[a]pyrene-induced genotoxicity
in human bronchial cells.
Offord EA; Macé K; Ruffieux C; Malnoë A; Pfeifer
Nestlé Research Centre, Lausanne, Switzerland.
Carcinogenesis, 16(9):2057-62 1995 Sep
The commonly used spice and flavouring agent, rosemary, derived
from the leaves of the plant Rosmarinus officinalis L., displays
antioxidant properties in foods and in biological systems. Moreover,
in animal models rosemary components were found to inhibit the
initiation and tumour promotion phases of carcinogenesis. In
this work, we studied the mechanisms by which rosemary components
block initiation of carcinogenesis by the procarcinogen benzo[a]pyrene
(B[a]P) in human bronchial epithelial cells (BEAS-2B). Whole
rosemary extract (6 micrograms/ml) or an equivalent concentration
of its most potent antioxidant constituents, carnosol or carnosic
acid, inhibited DNA adduct formation by 80% after 6 h co-incubation
with 1.5 muM B[a]P. Under similar conditions, cytochrome P450
(CYP) 1A1 mRNA expression was 50% lower in the presence of rosemary
components, and CYP1A1 activity was inhibited 70-90%. The observed
reduction of DNA adduct formation by rosemary components may
mostly result from the inhibition of the activation of benzo[a]pyrene
to its ultimate metabolites. Carnosol also affected expression
of the phase II enzyme glutathione-S-transferase which is known
to detoxify the proximate carcinogenic metabolite of B[a]P. Treatment
of BEAS-2B cells with carnosol (1 microgram/ml) for 24 h resulted
in a 3- to 4-fold induction of GST pi mRNA. Moreover, expression
of a second important phase II enzyme, NAD(P)H: quinone reductase,
was induced by carnosol in parallel with GST pi. Therefore, rosemary
components have the potential to decrease activation and increase
detoxification of an important human carcinogen, identifying
them as promising candidates for chemopreventive programs.
Flavonoids in Rosmarinus officinalis leaves.
Okamura N; Haraguchi H; Hashimoto K; Yagi A
Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University,
Phytochemistry, 37(5):1463-6 1994 Nov
Three new flavonoid glucuronides, luteolin 3'-O-beta-D-glucuronide,
luteolin 3'-O-(4"-O-acetyl)-beta-D-glucuronide, and luteolin
3'-O-(3"-O-acetyl)-beta-D-glucuronide, together with hesperidin,
were isolated from 50% aqueous MeOH extract of the leaves of
rosemary. The structures were established by chemical and spectroscopic
methods. Their antioxidant activities were evaluated by a ferric
thiocyanate method with hesperidin showing the greatest activity.
Inhibition of growth and aflatoxin production in Aspergillus
parasiticus by essential oils of selected plant materials.
Tantaoui-Elaraki A; Beraoud L
Department of Food Microbiology and Biotechnology, Hassan II
Institute for Agriculture and Veterinary Medicine, Rabat-Instituts,
J Environ Pathol Toxicol Oncol, 13(1):67-72 1994
We studied the effect of 13 chemically different essential oils
(EO) on the mycelial growth of and aflatoxin synthesis by Aspergillus
parasiticus. Cinnamon, thyme, oregano, and cumin EO were able
to stop mycelial growth at only 0.1% in the medium, while curcumin,
ginger, lemon, and orange EO were unable to inhibit totally the
growth even at 1% concentration. Coriander, black pepper, mugwort,
bay, and rosemary EO caused the growth to stop at concentrations
between 0.2 and 1%. The EO most active upon mycelial growth were
also the most active against aflatoxinogenesis. However, aflatoxin
synthesis was inhibited by all the EO at higher extent than the
Salicylates in foods.
Swain AR; Dutton SP; Truswell AS
J Am Diet Assoc, 85(8):950-60 1985 Aug
To determine salicylate content, 333 food items were analyzed.
Foods were homogenized with 25% sodium hydroxide, allowed to
stand overnight, acidified with concentrated hydrochloric acid,
and then extracted with warm diethylether over 5 hours. The extract
was dried and taken up in dilute sodium bicarbonate solution
for analysis. Salicylic acid was separated by high performance
liquid chromatography and quantified by reading at 235 nm. Salicylic
acid standards were used throughout to standardize extractions
and analyses. This is the most comprehensive set of data on food
salicylates yet published; extraction appears to have been more
complete for some foods, giving higher values than those previously
published. Most fruits, especially berry fruits and dried fruits,
contain salicylate. Vegetables show a wide range from 0 to 6
mg salicylate per 100 gm food (for gherkins). Some herbs and
spices were found to contain very high amounts per 100 gm, e.g.,
curry powder, paprika, thyme, garam masala, and rosemary. Among
beverages, tea provides substantial amounts of salicylate. Licorice
and peppermint candies and some honeys contain salicylates. Cereals,
meat, fish, and dairy products contain none or negligible amounts.
Mechanisms involved in the chemoprotective effects of rosemary
extract studied in human liver and bronchial cells.
Offord EA; Macé K; Avanti O; Pfeifer AM
Nestlé Research Centre, Lausanne, Switzerland. firstname.lastname@example.org
Cancer Lett, 114(1-2):275-81 1997 Mar 19
Natural polyphenols found in rosemary have not only potent antioxidant
activities but also anticarcinogenic properties. We have studied
some of the molecular mechanisms involved in their chemopreventive
action using in vitro human liver and bronchial cell models.
rosemary extract, or its active components, carnosol or carnosic
acid are potent inhibitors of DNA adduct formation induced by
benzo(a)pyrene or aflatoxin B1. At least two mechanisms are involved
in the anticarcinogenic action of rosemary extract: (i) inhibition
of the metabolic activation of procarcinogens catalysed by the
phase I cytochrome P450 enzymes; (ii) induction of the detoxification
pathway catalysed by the phase II enzymes such as glutathione
An evaluation of the antioxidant and antiviral action of extracts
of rosemary and Provençal herbs.
Aruoma OI; Spencer JP; Rossi R; Aeschbach R; Khan A; Mahmood
N; Munoz A; Murcia A; Butler J; Halliwell B
Pharmacology Group, University of London King's College, UK.
Food Chem Toxicol, 34(5):449-56 1996 May
Extracts of herbs and spices are increasingly of interest in
the food industry because they retard oxidative degradation of
lipids. There is also increasing interest in the antiviral activity
of plant products. A liquid, deodorized rosemary extract and
an oily extract of a mixture of Provençal herbs were tested
for antioxidant and antiviral action in vitro. The rosemary extract
(Herbor 025) and the extract of Provençal herbs (Spice
Cocktail) inhibited peroxidation of phospholipid liposomes with
50% inhibition concentration values of 0.0009% (v/v) and 0.0035%
(v/v), respectively. Herbor 025 and the spice cocktail (at 0.2%,
v/v) reacted with trichloromethylperoxyl radical with calculated
rates of 2.7 x 10(4) s-1 and 1.5 x 10(3) s-1, respectively. The
main active components in the herbal preparations, carnosol and
carnosic acid, at 0.05% (v/v) react with rate constants of (1-3)
x 10(6) M-1 sec-1 and 2.7 x 10(7) M-1 sec-1, respectively. Both
extracts show good antioxidant activity in the Rancimat test,
especially in lard. Herbor 025 and the spice cocktail inhibited
human immunodeficiency virus (HIV) infection at very low concentrations
which were also cytotoxic. However, purified carnosol exhibited
definite anti-HIV activity at a concentration (8 microM) which
was not cytotoxic. Both preparations promoted some DNA damage
in the copper-phenanthroline and the bleomycin-iron systems.
The two herbal preparations possess antioxidant properties that
may make them useful in the food matrix.
Study of the embryotoxic effects of an extract of rosemary (Rosmarinus
Lemonica IP; Damasceno DC; di-Stasi LC
Departamento de Farmacologia, Universidade Estadual Paulista,
Braz J Med Biol Res, 29(2):223-7 1996 Feb
Extracts of rosemary, Rosmarinus officinalis L., have been used
in folk medicine as a diuretic, an emenagogue, an antispasmodic
and its aqueous extract does not present toxicity to man, presenting,
however, abortive effects. In order to evaluate if this plant
induces abortion and/or interferes with the normal development
of the concepts, doses of 26 mg of a 30% (w/v) R. officinalis
aqueous extract (13 mg solids/ml) made with leaves, flowers and
stem were administered daily by gavage during two different periods
of Wistar rat pregnancy. One group of animals (N = 12) received
the extract from days 1 to 6 of pregnancy (preimplantation period)
and another group (N = 14) received the same extract from days
6 to 15 of pregnancy (organogenic period). Control groups (N
= 12) received saline in the same volume and during the same
periods as their respective experimental groups. The animals
were sacrificed at term. The treatment of the dams during either
the preimplantation or the organogenic period did not cause significant
changes in the postimplantation loss or in the number of anomalies
or malformations of the term fetuses, which also showed a similar
degree of development when compared with the respective controls.
The percent of preimplantation loss in the group treated before
embryo implantation increased, although the difference was not
significant compared to the control. This result suggests that
rosemary extract may present an anti-implantation effect without
interfering with the normal development of the concept after
Antioxidant and pro-oxidant properties of active rosemary constituents:
carnosol and carnosic acid.
1. Carnosol and carnosic acid have been suggested to account
for over 90% of the antioxidant properties of rosemary extract.
2. Purified carnosol and carnosic acid are powerful inhibitors
of lipid peroxidation in microsomal and liposomal systems, more
effective than propyl gallate. 3. Carnosol and carnosic acid
are good scavengers of peroxyl radicals (CCl3O2.) generated by
pulse radiolysis, with calculated rate constants of 1-3 x 10(6)
M-1 s-1 and 2.7 x 10(7) M-1 s-1 respectively. 4. Carnosic acid
reacted with HOCl in such a way as to protect the protein alpha
1-antiproteinase against inactivation. 5. Both carnosol and carnosic
acid stimulated DNA damage in the bleomycin assay but they scavenged
hydroxyl radicals in the deoxyribose assay. The calculated rate
constants for reaction with .OH in the deoxyribose system for
carnosol and carnosic acid were 8.7 x 10(10) M-1 s-1 and 5.9
x 10(10) M-1 s-1 respectively. 6. Carnosic acid appears to scavenge
H2O2, but it could also act as a substrate for the peroxidase
system. 7. Carnosic acid and carnosol reduce cytochrome c but
with a rate constant significantly lower than that of O2(-.).
Natural antioxidants as inhibitors of oxygen species induced
Minnunni M; Wolleb U; Mueller O; Pfeifer A; Aeschbacher HU
Nestec Ltd, Nestlé Research Centre, Vers-chez-les-Blanc,
Mutat Res, 269(2):193-200 1992 Oct
A ternary antioxidant vitamin mix consisting of ascorbic acid,
alpha-tocopherol and lecithin as well as a rosemary extract with
carnosic acid and carnosol as the two major active ingredients
were shown to exhibit strong antimutagenic effects in Ames tester
strain TA102. This strain has been shown to be highly sensitive
to reactive oxygen species. Mutagenicity was induced by the generation
of oxygen radicals by tert-butyl-hydroperoxide (tBOOH) or hydrogen
peroxide (H2O2); therefore, the antimutagenic property of the
above substances was attributed to their antioxidant properties.
In the case of the vitamin mix, ascorbic acid was held responsible
for this inhibitory property, whereas for the rosemary extract
carnosic acid was identified as the antimutagenic agent. Since
oxygen radicals are known to be involved in the multiprocess
of carcinogenicity, it is concluded that these antioxidants might
exhibit anticarcinogenic properties.
Effects of three dietary phytochemicals from tea, rosemary and
turmeric on inflammation-induced nitrite production.
Radical intermediates and antioxidants: an ESR study of radicals
formed on carnosic acid in the presence of oxidized lipids.
Geoffroy M; Lambelet P; Richert P
Department of Physical Chemistry, University of Geneva, Switzerland.
Free Radic Res, 21(4):247-58 1994 Sep
Carnosic acid, an antioxidant extracted from rosemary, is shown
to produce radicals when in contact with oxidized methyl oleate
in the absence of air above 50 degrees C. Two radical species
are formed: the first one, stable up to approximately 110 degrees
C, is an hydroxy-phenoxy radical whose ESR spectrum was analyzed
by studying its temperature dependence and its sensitivity to
deuterium/proton exchange. The second species was observed above
110 degrees C, its ESR spectrum was identical to the spectrum
obtained when carnosol, another antioxidant extracted from rosemary,
was heated at the same temperature in the presence of oxidized
lipid. This observation is probably due to the transformation
of carnosic acid into carnosol; the analysis of the corresponding
ESR spectrum suggests the formation of a keto phenoxy radical
exhibiting a great delocalization of the unpaired electron.
Inhibitory effect of carnosic acid on HIV-1 protease in cell-free
assays [corrected] [published erratum appears in J Nat Prod 1994
Paris A; Strukelj B; Renko M; Turk V; Pukl M; Umek A; Korant
Department of Biochemistry, Jozef Stefan Institute, Ljubljana,
J Nat Prod, 56(8):1426-30 1993 Aug
In order to find new effective HIV protease inhibitors, two diterpenes
(carnosic acid  and carnosol ) were isolated from rosemary
(Rosmarinus officinalis L.), and rosmanol  and semisynthetic
derivatives (7-O-methylrosmanol , 7-O-ethylrosmanol , and
11,12-O,O-dimethylcarnosol ) were prepared. The inhibitory
activity of all six compounds against HIV-1 protease was tested.
The carnosic acid  showed the strongest inhibitory effect
(IC90 = 0.08 micrograms/ml). The same compound was also assayed
against HIV-1 virus replication (IC90 = 0.32 micrograms/ml).
The cytotoxic TC90 on H9 lymphocytes was 0.36 micrograms/ml,
which is very close to the effective antiviral dose. Additionally,
the tested compounds did not inhibit cellular aspartic proteases
cathepsin D and pepsin at the concentration range up to 10 micrograms/ml