Nenhuma entrada de sumrio foi encontrada.

INTRODUO
Curcuma longa L., which belongs to the Zingiberaceae family, is a perennial herb that
measures up to 1 m high with a short stem, distributed throughout tropical and subtropical
regions of the world, being widely cultivated in Asiatic countries, mainly in India and China.
The major constituent, curcumin (difer-uloylmethane) is in the most important fraction of C.
longa L. and its chemical structure, was determined by Roughley and Whiting (1973). It melts
at 176-177C and forms red-brown salts with alkalis. Curcumin is soluble in ethanol, alkalis,
ketone, acetic acid and chloroform; and is insoluble in water. In the molecule of curcumin, the
main chain is aliphatic, unsaturated and the aryl group can be substituted or not.
Anti-inflammatory activity - There is a great number of papers in the literature relating the
activity of compounds extracted from C. longa L. being potent inhibitors of inflammation.
These substances can be classified as curcuminoids, analogues of diarylheptanoids. There
are two models of inflammation to be studied: chronic models (cotton pellet and granuloma
pouch), where the inflammation and granulomas develop during a period of time (several
days), indicating the proliferative phase of inflammation; and acute models, where acute
effects of anti-inflammatory agents can be studied, testing their inhibitory action on the
development of rat paw edema.
Antioxidant activity - Unnikrishnan and Rao (1995) studied the antioxidative properties of curcumin
and its three derivatives (demethoxy curcumin, bisdemethoxy curcumin and diacethyl curcumin). The
authors demonstrated that these substances provide a protection of hemoglobin from oxidation at a
concentration as low as 0.08 mM, except the diacethyl curcumin which has little effect in the inhibition
of nitrite induced oxidation of hemoglobin.
The effect of curcumin on lipid peroxidation has also been studied in various models by
several authors. Curcumin is a good antioxidant and inhibits lipid peroxidation in rat liver
microsomes, erythrocyte membranes and brain homogenates (Pulla Reddy & Lokesh 1994).
The lipid peroxidation has a main role in the inflammation, in heart diseases, and in cancer.
Anti-tumor activity - Huang et al. (1988), studying the effect of curcumin, chlorogenic acid,
caffeic acid and ferulic acid on tumor promotion in mouse skin by 12-O-tetradecanoyl-13-
acetate (TPA), observed that all these compounds inhibit the epidermal ornithine
decarboxilase (ODC) and epidermal DNA synthesis, being curcumin the most efficient. In
another work (1991), the results suggested that curcumin was a potent inhibitor of TPA- and
arachidonic acid-induced inflammation and of lipoxygenase and cyclooxygenase activities in
mouse epidermis. The IC
50
for curcumin-dependent inhibition of these enzyme activities was
5-10 M. In this study the results indicated that curcumin inhibited the epidermal metabolism
of arachidonic acid via the lipoxygenase and cyclooxygenase pathways.
Furthermore, Ozaki et al. (2000), examining the action of curcumin on rabbit osteoclast
apoptosis, demonstrated that curcumin drastically inhibits bone resorption in parallel with its
stimulation of apoptosis in the cells. Since cancer and bone inflammation are diseases that
increase bone resorption, the authors suggest that curcumin may be useful in the therapy of
these pathogenies.
MATERIALS AND METHODS
Reagents
AlamarBlue was purchased from Biosource (Camarillo - CA, USA, cat.# DAL1025),
curcumin (cat.# C1386) was acquired from Sigma (St. Louis - MO, USA) and TPP 96-well
tissue culture microplates were from TPP (Trasadingen, Switzerland). DMEM media
supplemented with 100 U/mL of penicillin and 100 g/mL of streptomycin, serum fetal
bovine, and Trypsin-EDTA solution were purchased from CultiLab (Campinas - SP, Brazil).
Curcumin Cytotoxicity Assay
In the cytotoxicity assay, 5x104 confluent monolayer adherent MB49 cells grown in
DMEM medium supplemented with 10% of serum fetal bovine, 100 U/mL of penicillin,
and 100 g/mL of streptomycin were seeded in a 96-well tissue culture microplate and
maintained for 24 hours at 37oC in a 5% CO2 atmosphere. Thus, cells were exposed to
different concentrations of curcumin (from 6 uM up to 400 uM) diluted in a fresh
supplemented media for 20 hours in triplicate. After two washings with PBS1x, cells were
incubated in fresh media containing 10% of AlamarBlue (resazurin) for three hours, and
the reduction level of resazurin was quantified using the microplate reader UVM340
(ASYS Hitech, Eugendorf - Austria) which measured the difference between the density
optics of wavelengths 570 and 600 um. As a 0% viability control (OD
min
), triton X 100
(1% of final concentration) was added to the cells one hour before the changing of
media. The complete media without curcumin was used as 100% viability control
(OD
max
). This experiment was repeated twice.
Animal Model
The technique of the establishment of animal tumor model was already described by
Chade et al. (10), briefly: Eight to ten-week-old female C57BL/6 mice, weighing 15-20g,
were provided by the University of Sao Paulo and maintained at our animal care facility
for one week prior to use. The mice were housed in groups of five per cage in a limited-
access area at a controlled room temperature, with food and water ad libitum. The
experiments were approved by the institution's ethical board council.
Orthotopic Tumor Implantation
Six- to eight-week-old female C57BL/6 mice were administered general anesthesia with
i.p. injection of a mixture of xylazine-ketamine (0.1 mL/10g/mouse). Then, a 24-gauge
Teflon i.v. catheter (Nipro Medical Ltda, Sorocaba, SP, Brazil) was inserted through the
urethra into the bladder using an inert lubricant (sterile contact gel). In order to prepare
the bladder for tumor implantation, a chemical lesion on the bladder wall was made by
intravesical instillation of 0.3 M AgNO
3
(8 L). This promoted an adequate and controlled
diffuse bladder wall cauterization. After 10 seconds, the content was washed out by
transurethral infusion of PBS. Then, a suspension of 1 x 10
5
viable tumor cells was
instilled into the bladder.