Printer Friendly

Analgesic and anti-inflammatory activities of a fraction rich in oncocalyxone A isolated from Auxemma oncocalyx.

Summary

In the present work we studied the antinociceptive and antiedematogenic effects of a quinone fraction (QF) isolated from the heartwood of Auxemma oncocalyx Taub. The major constituent of QF, which represented around 80% of this fraction, was a terpenoid quinone named oncocalyxone A (1). Results show that QF (10 and 30 mg/kg body wt., i.p.) significantly inhibited paw edema induced by carrageenan at the second, third, and fourth hours. The effect was dose-dependent and long lasting, and QF was less effective orally. An antiedematogenic effect was also demonstrated in the dextran-induced paw edema. In this model, however, QF was somewhat less potent. QF (1 and 5 mg/kg body wt., i.p.) inhibited acetic acid-induced abdominal contractions in mice in a dose-dependent manner. In addition, QF (5 and 10 mg/kg body wt., i.p.) inhibited only the second phase (inflammatory) in the formalin test, and showed no effect in the hot-plate test in mice. The antinociceptive activity of QF was predominantly peripheral and independent of the opioid system. The observed effects of QF are, at least in part, probably due to the presence of oncocalyxone A (1).

Key words: Analgesic and antiinflammatory activities, oncocalyxone A, Auxemma oncocalyx, quinones

**********

Introduction

Auxemma oncocalyx Taub, belongs to the Boraginaceae family and is native to the northeastern Brazilian "caatinga", where it is known as "pau branco". The stem bark of the tree is astringent and used popularly in the treatment of wounds (Braga, 1976; Pessoa, 1994). Alantoin isolated from the stem hydroalcoholic extract (Pessoa and Lemos, 1997) explains partially the beneficial use of the plant in wound healing. Several other compounds were also isolated from the hydroalcoholic extract, including [beta]-sitosterol, its glycoside (3[beta]-O-D-glucopyranosylsitosterol) and seven terpenoidic quinones, among them oncocalyxone A, 1 (Pessoa et al. 1993; Pessoa et al. 1995).

Previous pharmacological studies showed that the stem hydroalcoholic extract possesses anti-platelet activity (Fontenele and Sousa, 1992) and this effect was confirmed recently in a hydrosoluble fraction, obtained from the heartwood methanolic extract, which also presented antioxidant activity (Ferreira et al. 1999; Ferreira et al. 2001). Other effects were also attributed to the hydroalcoholic extract, such as antitumoral (Pessoa et al. 1992), analgesic and anti-inflammatory (Lino et al. 1996) activities. Recently, in vitro studies demonstrated the antitumoral activity not only of oncocalyxone A (1), but also of oncocalyxone C, isolated from A. oncocalyx (Pessoa et al. 2000; Leyva et al. 2000).

Quinones show several types of biological activity, including, platelet antiaggregant (Teng et al. 1993; Chung et al. 1994), antioxidant (Belisario et al. 1992; Houghton et al. 1995; Tripathi et al. 1995; Mori et al. 1998), anti-inflammatory (Kuo et al. 1995); Vazquez et al. 1996; Odukoya et al. 1999), analgesic (Hernandez-Perez et al. 1995; Abdel-Fattah et al. 2000), antitumoral (Morello et al. 1995; Itoigawa et al. 2000), antifungal (Perry et al. 1991; Gafner et al. 1996), antimalarial (Figueiredo et al. 1998), and leishmanicidal (Sauvain et al. 1993; Sittie et al. 1999). The objectives of the present work were to study the analgesic and anti-inflammatory effects of the quinone-enriched fraction from A. oncocalyx and to elucidate its possible mechanism of action.

Materials and Methods

Plant material and fractionation

The plant was collected at the city of Pentecoste, state of Ceara, in northeastern Brazil, and identified by Prof. A.G. Fernandes of the Biology Department of the Federal University of Ceara. A voucher specimen (No. 18459) has been deposited at the Prisco Bezerra Herbarium. The quinone fraction (QF) was prepared from the ground heartwood methanolic extract through exhaustive aqueous extraction followed by lyophilization. The hydrosoluble fraction contained around 80% of oncocalyxone A (1) (Fig. 1), which was previously characterized by spectroscopy analysis as MS, IR and NMR including 2D sequences such as COSY, HETCOR and COLOC (Pessoa et al. 1993). The concentration of (1) in the hydrosoluble fraction was calculated on the basis of the HPLC-fingerprint analysis using YMC-Pack C-18 column, acetonitrile: [H.sub.2]O (1:1, v/v) as mobile phase with an isocratic elution program at room temperature and detection at 280 nm.

[FIGURE 1 OMITTED]

Test animals

Male Wistar rats (150-200 g each) and male Swiss mice (25-30 g each) were obtained from the Animal House of the Federal University of Ceara. Animals were maintained in an air-conditioned room at 23-25[degrees]C, 12 h light-12 h dark cycle, and fed with a standard laboratory diet and tap water ad libitum. Experiments were performed according to the Guide for use and care of laboratory animals of the Department of Health and Human Services of the United States of America.

Drugs

Carrageenan, dextran and naloxone were purchased from Sigma, U.S.A. Morphine sulfate was obtained from Cristalia, Brazil. All other drugs were of analytical grade.

Pharmacological tests

Carrageenan-induced paw edema (Winter et al. 1962): Male Wistar rats (150-200 g each) divided into groups of 6 animals each were used. QF was administered at doses of 1, 10 and 30 mg/kg body wt., i.p., 30 min before the intraplantar injection of a 1% carrageenan solution (0.1 ml in the right hind paw). Paw volume was measured using a plethysmometer (Ugo Basile, Italy), before and at 1;2;3;4 and 24 h after carrageenan administration. The difference between the paw volume before and after carrageenan administration was considered as an index of edema formation at each observation time. Controls received saline (10 ml/kg body wt., i.p.). QF (100 and 200 mg/kg body wt.) was also administered orally, 60 min before injection of carrageenan, and in this case controls received a 2% DMSO solution in saline (used as vehicle for QF) and animals were fasted for approximately 12 h.

Dextran-induced paw edema (Parrat and West, 1958): Male Wistar rats (150-200 g each) divided into 6 animals per group were used. QF was administered at doses of 1, 10 and 30 mg/kg body wt., i.p., 30 min before the intraplantar injection of 0.1 ml of 1.5% dextran solution in the right hind paw. The paw volume was measured using a plethysmometer before and 1/2; 1; 2; 3 and 4 h after dextran injection. The difference between the paw volume before and after dextran injection is considered as an index of edema at each observation time. Controls received saline (10 ml/kg body wt., i.p.).

Acetic acid-induced abdominal contractions (Koster et al. 1959): Male Swiss mice (20-30 g each), divided into groups of 10 animals each were used. The quinone fraction (QF) was administered once at doses of 0.1, 1 and 5 mg/kg body wt., i.p., 30 min before the 0.6% acetic acid injection (10 ml/kg body wt., i.p.). Ten minutes later, the number of abdominal contractions was registered for 20 min. The control group received saline, used as vehicle for QF.

Formalin test (Hunskaar et al. 1985; Tjolsen et al. 1992); Male Swiss mice (20-30 g each) divided into groups of 6 animals each were used. QF was administered once at doses of 1, 5, 10 and 30 mg/kg body wt., i.p., 30 min before the intraplantar injection of 20 [micro]l of 1% formalin in the right hind paw. The licking time (in seconds) was registered at 5 min (first phase, neurogenic), and after 20 min for 5 min (second phase, inflammatory), after formalin injection. The control group received saline, 10 ml/kg body wt., i.p. Morphine was used as a standard (5 mg/kg body wt., i.p.). In order to verify the possible involvement of the opioid system in the analgesic effect of QF, the formalin test was performed also in the presence of the opioid antagonist, naloxone (2 mg/kg body wt., subcutaneously). QF (30 mg/kg body wt., i.p.) and morphine were administered 15 min after and 30 min before naloxone and formalin injections, respectively.

Hot-plate test (Eddy and Leimback, 1953): Male Swiss mice (20-30 g each) divided into groups of 10 animals each were used. Animals were selected previously from those presenting a latency to the thermal stimulus equal to or less than 20 sec; and the cut off point was set at 40 sec. QF was administered once at doses of 5, 10 and 30 mg/kg body wt., i.p. Latency to the thermal stimulus was registered before and at 30, 60 and 90 min after QF administration. The control group received saline (10 ml/kg body wt., i.p.). Morphine (5 mg/kg body wt., i.p.) was used as a standard.

Statistical analysis

Data were analyzed by one-way Analysis of Variance (ANOVA) followed by the Tukey-Kramer test for multiple comparisons.

Results

The quinone fraction was very active in reducing the rat paw edema after carrageenan administration (Table 1). The results show inhibitions of approximately 46, 57 and 48%, at the second, third and fourth hours, respectively, after the dose of 10 mg/kg body wt., i.p. and of 50, 60 and 51% at the same intervals following the dose of 30 mg/kg body wt., i.p. The effect was long-lasting and reductions of 38% in the edema could still be seen after a 24-h period (with the highest dose). Although less potent, QF was also effective after oral administration (100 and 200 mg/kg body wt.); however, under these conditions, decreases of edema ranging from 25 to 29% were observed at the third, fourth and twenty-fourth hours after carrageenan administration, as compared to controls.

Data in Table 2 show the effect of QF (1, 10 and 30 mg/kg body wt., i.p.) on dextran-induced paw edema in rats. Significant reductions of paw edema were observed at the dose of 30 mg/kg body wt., i.p., at the first (27%), second (45%), third (48%) and fourth (42%) hours. QF was active at the dose of 10 mg/kg body wt., i.p., only at the second (36%), third (42%) and fourth (33%) hours.

Table 3 shows the effects of QF on acetic acid-induced abdominal contractions in mice. QF (1 and 5 mg/kg body wt., i.p.) inhibited abdominal contractions by 26 and 60%, respectively, as compared to controls (24.5 [+ or -] 1.8 contractions/20 min). No effect was observed, however, at the lower dose of QF (0.1 mg/kg body wt., i.p.).

Table 4 shows the effects of QF on formalin-induced nociception in mice. QF at doses of 5, 10 and 30 mg/kg body wt., i.p., significantly inhibited licking time in the second phase of the response by 27, 52 and 62%, respectively, as compared to controls. No effect was observed in the first phase of the response. On the other hand, morphine inhibited licking time in the first and second phases of the response by 55 and 85%, respectively. Although the effects of morphine were totally reversed, no change was seen in the effect of QF in the presence of naloxone, as compared to the effect of QF alone.

QF at doses of 5, 10 and 30 mg/kg body wt., i.p. did not, however, alter latency of reaction to the thermal stimulus, as demonstrated by the hot-plate test. Morphine, used as positive control, produced increases on the order of 84, 57 and 65% after 30, 60 and 90 min, respectively, as compared to control (Table 5).

Discussion

The present work showed that QF, administered intraperitoneally or orally, inhibited carrageenan-induced rat paw edema in a dose-dependent manner. The maximum effect was observed even at the dose of 10 mg/kg body wt., i.p. The inhibition was long-lasting, still observable 24 h after carrageenan administration. The inhibition was significant at the second, third and fourth hours, when bradykinin is being released and there occurs an accumulation of prostaglandins and infiltration of polymorphonuclear cells. At this stage, there is also production of free radicals, among other mediators. The antiedematogenic effect of QF was greater after intraperitoneal administration, indicating that oral absorption is less efficient. This effect was also seen in the dextran-induced paw edema test, where QF inhibited the edema dose-dependently after intraperitoneal administrations of 10 and 30 mg/kg body wt.

Earlier research showed that in carrageenan-induced edema, several mediators (histamine, 5-hydroxytryptamine, kinins, prostaglandins) of the inflammatory process are involved, including the complement system (Willis, 1969; Willoughby et al. 1969). Di Rosa et al. (1971) showed that this system participates in the process from the very beginning, and that carrageenan-induced inflammation is divided into 3 steps according to the mediators released. In the initial step (first 90 min), the release of both histamine and 5-hydroxytryptamine (5-HT) occurs; and the second step (from 90 to 150 min) is mediated by kinins, while in the third step (from 150 min on), prostaglandin release takes place. All these mediators are dependent upon the complement system. Histamine and 5-HT are responsible for vasodilation and increase in vascular permeability in the initial phase of the inflammatory process. Bradykinin has been implicated in acute inflammatory processes due to its ability to induce an increase in blood vessel permeability. Its involvement in carrageenan-induced edema was demonstrated by Ronald and Christopher (1990) and by Damas and RemacleVolon (1992). Another characteristic of carrageenan-induced edema is the massive infiltration of polymorphonuclear leucocytes observed in the third step (Di Rosa and Willoughby, 1971; Ialenti et al. 1992; Masso et al. 1993). By contrast, dextran-induced edema involves mast cell degranulation, resulting in histamine as well as 5-HT release.

Histamine, 5-HT and bradykinin are able to induce nitric oxide (NO) release from vascular endothelial cells in vitro via a mechanism involving receptor occupation and stimulation of NO synthase. In addition, macrophages produce NO when activated by lipopolysaccharide or cytokines. Thus Salvemini et al. (1996) pointed to NO as an important mediator of carrageenan-induced edema and suggested that constitutive NO synthase (cNOS) acts at the initial steps, while inducible NO synthase (iNOS) is involved in the last step of the inflammatory reaction.

The acute or chronic inflammatory process is a model of nociception (Besson, 1997) and several works have shown that inflammatory mediators can generate nociceptive impulses (Steen et al. 1996; Besson, 1997). Thus, bradykinin and 5-HT are able to stimulate cutaneous nociceptors and 5-HT can also increase nociceptive response to bradykinin (Lang et al. 1990; Rueff and Dray, 1993). Although prostaglandins are not able to activate these nociceptors, they promote sensitization of these same nociceptors to bradykinin in some tissues (Schaible and Schmidt, 1988). Additionally, neuropeptides such as substance P play a role in neurogenic inflammation (Steen et al. 1996). Recent evidence showed the importance of cytokines, including tumor necrosis factor [alpha] (TNF[alpha]), in the generation and maintenance of hyperalgesia associated with neuropathic pain (Jungler and Sorkin, 2000).

Considering the relationship between anti-inflammatory and analgesic effects, another objective of the present work was to study the antinociceptive activity of QF using the writhing and formalin test in mice. Acetic acid produces a painful reaction and acute inflammation in the peritoneal area. The stimulation of peritoneal nociceptors is indirect and occurs through the release of endogenous substances, which stimulate nervous endings (Gyires and Torna, 1984). A great increase occurs in prostaglandins [E.sub.2] and [F.sub.2a] levels in peritoneal fluid after acetic acid injection, and the analgesic effect of substances similar to aspirin could be due to the blockade of prostaglandin synthesis.

QF was able to significantly and dose-dependently inhibit the abdominal contractions induced by acetic acid, as well as the second phase of the response to formalin. The formalin test is considered a model for chronic pain (Dubuisson and Dennis, 1977). In this test, animals present two distinct nociceptive behavior phases, which probably involve different stimuli. The first phase initiates immediately after formalin injection and lasts 3 to 5 min, resulting from chemical stimulation of nociceptors. The second phase initiates 15 to 20 min after formalin injection, lasts 20 to 40 min and seems to depend on a peripheral mechanism as well as a central one. While substance P and bradykinin are involved in the first phase, histamine, 5-HT, prostaglandins and bradykinin are involved in the second phase. The effect of QF was significant only in the second phase, indicating an action related to the inflammatory process. This QF effect was not reversed by naloxone, pointing to the non-involvement of the opioid system. Finally, QF showed no effect in the hotplate test, which involves higher brain functions and consists of responses to nociceptive stimuli organized at a supraspinal level (Gardmark et al. 1998). Analgesic drugs such as aspirin and paracetamol do not have any effect in this test (Tjolsen et al. 1992), while it is largely used to measure opioid effects (Gong et al. 1991; Plone et al. 1996).
Table 1. Effect of the quinone fraction (QF) of Auxemma oncocalyx Taub.
on carrageenan-induced paw edema in rats.

Group (n) Edema volume in ml (% Inhibition)
 1 h 2 h

Control, i.p. (6) 0.61 [+ or -] 0.03 1.03 [+ or -] 0.10
QF
 1 mg/kg body 0.59 [+ or -] 0.06 0.98 [+ or -] 0.09
 wt., i.p. (6) (3.3) (4.9)
 10 mg/kg body 0.45 [+ or -] 0.05 0.56 [+ or -] 0.05*
 wt., i.p. (6) (26.2) (45.6)
 30 mg/kg body 0.44 [+ or -] 0.04 0.52 [+ or -] 0.05*
 wt., i.p. (6) (27.9) (49.5)
Control, p.o. (6) 0.94 [+ or -] 0.06 1.54 [+ or -] 0.13
QF
100 mg/kg body 0.79 [+ or -] 0.14 1.57 [+ or -] 0.08
 wt., p.o. (6) (15.9) (-)
200 mg/kg body 0.73 [+ or -] 0.06 1.47 [+ or -] 0.18
 wt., p.o. (6) (22.3) (4.6)

Group (n) Edema volume in ml (% Inhibition)
 3 h 4 h

Control, i.p. (6) 1.62 [+ or -] 0.10 1.41 [+ or -] 0.13
QF
 1 mg/kg body 1.35 [+ or -] 0.17 1.28 [+ or -] 0.11
 wt., i.p. (6) (16.7) (9.2)
 10 mg/kg body 0.69 [+ or -] 0.06* 0.74 [+ or -] 0.06*
 wt., i.p. (6) (57.4) (47.5)
 30 mg/kg body 0.65 [+ or -] 0.05* 0.69 [+ or -] 0.05*
 wt., i.p. (6) (59.9) (51.0)
Control, p.o. (6) 2.06 [+ or -] 0.10 2.09 [+ or -] 0.11
QF
100 mg/kg body 1.70 [+ or -] 0.13 1.93 [+ or -] 0.09
 wt., p.o. (6) (17.5) (7.7)
200 mg/kg body 1.54 [+ or -] 0.16* 1.58 [+ or -] 0.12*
 wt., p.o. (6) (25.2) (24.4)

Group (n) Edema volume in ml (% Inhibition)
 24 h

Control, i.p. (6) 0.29 [+ or -] 0.04
QF
 1 mg/kg body 0.27 [+ or -] 0.03
 wt., i.p. (6) (6.9)
 10 mg/kg body 0.20 [+ or -] 0.02
 wt., i.p. (6) (31.0)
 30 mg/kg body 0.18 [+ or -] 0.02
 wt., i.p. (6) (37.9)
Control, p.o. (6) 0.24 [+ or -] 0.04
QF
100 mg/kg body 0.19 [+ or -] 0.02
 wt., p.o. (6) (20.8)
200 mg/kg body 0.17 [+ or -] 0.02
 wt., p.o. (6) (29.2)

Male Wistar rats (150-200 g each) were used. Control animals received
saline (10 ml/kg body wt.) and treated groups received QF
intraperitoneally or orally, 30 or 60 min, respectively, before the
intraplantar administration of 1% carrageenan solution (0.1 ml, right
hind paw). Edema volume (expressed in ml) was estimated at 1, 2, 3, 4
and 24 h after carrageenan administration. Values are expressed as means
[+ or -] S.E.M. of the number of animals. *p < 0.05 vs. control (One-way
ANOVA and Tukey-Kramer as post-hoc test).

Table 2. Effect of the quinone fraction (QF) of Auxemma oncocalyx Taub.
on dextran-induced paw edema in rats.

Group (n) Edema volume in ml (% Inhibition)
 1/2 h 1 h

Control, i.p. (6) 2.4 [+ or -] 0.13 3.0 [+ or -] 0.13
QF 1 mg/kg body 2.8 [+ or -] 0.21 3.1 [+ or -] 0.18
 wt., i.p. (6)
QF 10 mg/kg body 2.2 [+ or -] 0.11 2.4 [+ or -] 0.08
 wt., i.p. (6) (8.3) (20.0)
QF 30 mg/kg body 2.0 [+ or -] 0.30 2.2 [+ or -] 0.27*
 wt., i.p. (6) (16.7) (26.7)

Group (n) Edema volume in ml (% Inhibition)
 2 h 3 h

Control, i.p. (6) 3.1 [+ or -] 0.11 3.1 [+ or -] 0.08
QF 1 mg/kg body 3.0 [+ or -] 0.12 2.7 [+ or -] 0.14
 wt., i.p. (6)
QF 10 mg/kg body 2.0 [+ or -] 0.24* 1.8 [+ or -] 0.24*
 wt., i.p. (35.5) (41.9)
QF 30 mg/kg body 1.7 [+ or -] 0.23* 1.6 [+ or -] 0.28*
 wt., i.p. (6) (45.2) (48.4)

Group (n) Edema volume in ml (% Inhibition)
 4 h

Control, i.p. (6) 2.4 [+ or -] 0.15
QF 1 mg/kg body 2.7 [+ or -] 0.12
 wt., i.p. (6)
QF 10 mg/kg body 1.6 [+ or -] 0.25
 wt., i.p. (33.3)
QF 30 mg/kg body 1.4 [+ or -] 0.24*
 wt., i.p. (6) (41.7)

Male Wistar rats (150-200 g each) were used. Control animals received
saline (10 ml/kg body wt.) and treated groups received QF 30 min before
the intraplantar injection of 0.1 ml, 1.5% dextran solution in the right
hind paw. Edema volume (in ml) was estimated at 1/2, 1, 2, 3 and 4 h
after dextran administration. Values are expressed as means [+ or -]
S.E.M. of the number of animals. *p < 0.05 vs. control (One-way ANOVA
and Tukey-Kramer as post-hoc test).

Table 3. Antinociceptive effect of the quinone fraction (QF) of Auxemma
oncocalyx Taub. on acetic acid-induced abdominal contractions in mice.

 Number of
Group (n) contractions % Inhibition

Control, i.p. (16) 24.5 [+ or -] 1.80
QF 0.1 mg/kg body wt., i.p. (10) 23.6 [+ or -] 1.09
QF 1 mg/kg body wt., i.p. (10) 18.2 [+ or -] 1.42* 26
QF 5 mg/kg body wt., i.p. (10) 9.7 [+ or -] 0.92* .60

Male Swiss mice (20-30 g each) were used. Control animals received
saline (10 ml/kg body wt.) and treated groups received QF 30 min before
an 0.6% acetic acid injection (10 ml/kg body wt., i.p.). Ten minutes
later, the number of abdominal contractions was registered over 20 min.
Values are expressed as means [+ or -] S.E.M. of the number of animals
used in the test (in parentheses). *p < 0.05 vs. control (One-way ANOVA
and Tukey-Kramer as post-hoc test). Percentages of inhibition of
contraction number relative to control are also presented.

Table 4. Evaluation of the opioid system's involvement in the effect of
the quinone fraction (QF) of Auxemma oncocalyx Taub. on formalin-induced
nociception in mice.

Group Licking time (sec)
 first phase second phase

Control, i.p. (21) 56.6 [+ or -] 2.57 26.4 [+ or -] 1.86
QF 1 mg/kg body wt. (11) 55.1 [+ or -] 3.06 25.3 [+ or -] 1.26
QF 5 mg/kg body wt. (12) 53.5 [+ or -] 3.14 19.3 [+ or -] 1.68*
QF 10 mg/kg body wt. (10) 58.5 [+ or -] 2.73 12.8 [+ or -] 1.22*
Control, i.p. (12) 50.9 [+ or -] 2.63 28.2 [+ or -] 2.77
QF 30 mg/kg body wt. (11) 48.4 [+ or -] 3.24 10.7 [+ or -] 1.35*
Mor (12) 22.9 [+ or -] 1.75* 4.2 [+ or -] 0.89*
Nal + QF (10) 47.4 [+ or -] 3.34 9.1 [+ or -] 1.57*
Nal + Mor (12) 50.6 [+ or -] 4.41 25.6 [+ or -] 2.69

Group % Inhibition
 first phase second phase

Control, i.p. (21)
QF 1 mg/kg body wt. (11)
QF 5 mg/kg body wt. (12) 27
QF 10 mg/kg body wt. (10) 52
Control, i.p. (12)
QF 30 mg/kg body wt. (11) 62
Mor (12) 55 85
Nal + QF (10) 68
Nal + Mor (12)

Male Swiss mice (20-30 g each) were used. Control group received saline
(10 ml/kg body wt.) and treated groups received QF or morphine (5 mg/kg
body wt.) intraperitoneally, 30 min before the intraplantar injection of
20 [micro]l of 1% formalin in the right hind paw. The licking time (in
seconds) was registered at the first 5 min (first phase) and after 20
min over 5 min (second phase) after the formalin injection. In the test
using Naloxone (Nal) and the quinone fraction, QF was administered 15
min after naloxone (2 mg/kg body wt., subcutaneously) and 30 min before
formalin injection. Morphine was also administered 15 min after Naloxone
(Nal) and 30 min before formalin injection. Values are expressed as
means [+ or -] S.E.M. of the number of animals in parentheses. *p < 0.05
vs. control (One-way ANOVA and Tukey-Kramer as post-hoc test).
Percentages of inhibition of licking time relative to control are also
presented.

Table 5. Effect of the quinone fraction (QF) of Auxemma oncocalyx Taub.
on the hot-plate test in mice.

Group Latency time (sec)
 0 min 30 min

Control, i.p. (16) 15.8 [+ or -] 1.13 15.2 [+ or -] 0.96
QF 5 mg/kg body wt., i.p.
 (10) 13.6 [+ or -] 1.38 14.2 [+ or -] 1.06
QF 10 mg/kg body wt., i.p.
 (10) 12.5 [+ or -] 1.34 13.7 [+ or -] 1.74
QF 30 mg/kg body wt., i.p.
 (10) 16.0 [+ or -] 1.51 16.2 [+ or -] 1.04
Morphine 5 mg/kg body wt.,
 i.p. (10) 15.3 [+ or -] 1.54 28.0 [+ or -] 3.04*

Group Latency time (sec)
 60 min 90 min

Control, i.p. (16) 13.5 [+ or -] 0.87 12.6 [+ or -] 1.10
QF 5 mg/kg body wt., i.p.
 (10) 10.9 [+ or -] 0.57 13.3 [+ or -] 1.28
QF 10 mg/kg body wt., i.p.
 (10) 11.2 [+ or -] 1.37 11.8 [+ or -] 1.16
QF 30 mg/kg body wt., i.p.
 (10) 15.8 [+ or -] 0.87 12.6 [+ or -] 1.29
Morphine 5 mg/kg body wt.,
 i.p. (10) 23.7 [+ or -] 2.09* 19.3 [+ or -] 1.68*

Male Swiss mice (20-30 g each) were used. Control animals received
saline (10 ml/kg body wt.) and treated groups received QF or morphine.
The latency to the thermal stimulus was registered (in seconds) before
(0 min) and at 30, 60 and 90 min after QF administration. Values are
expressed as means [+ or -] S.E.M. of the number of animals in
parentheses. *p < 0.01 vs. control (One-way ANOVA and Tukey-Kramer as
post-hoc test).


Acknowledgements

The authors wish to thank the Brazilian National Research Council (CNPq) for supporting this work and Maria V. R. Bastos for technical assistance.

References

Abdel-Fattah AM, Matsumo K, Watanabe H (2000) Antinociceptive effects of Nigella sativa oil and its major component, thymoquinone, in mice. Eur J Pharmacol 400: 89-97

Belisario MA, Maturo M, Pecce R, De Rosa S, Villani GR (1992) Effect of avarol and avarone on in vitro-induced microsomal lipid peroxidation. Toxicology 72: 221-233

Besson JM (1997) The complexity of physiopharmacological aspects of pain. Drugs 5: 1-9

Braga R (1976) Plantas do Nordeste, especialmente do Ceara. Mossoro: Editora Universitaria da UFRN, 540 p. (Colecao Mossoroense, 3)

Chung MI, Gan KH, Lin CN, Ko FN, Teng CM (1994) Antiplatelet effects and vasorelaxing action of some constituents of formosan plants. J Nat Prod 57: 313-316

Damas J, Remacle-Volon, G (1992) Influence of a long-acting bradykinin antagonist, Hoe 140, on some acute inflammatory reactions in the rat. Eur J Pharmacol 211: 81-86

Di Rosa M, Giroud JP, Willoughby DA (1971) Studies of the mediators of the acute inflammatory response induced in rats in different sites by carrageenan and turpentine. J Pathol 104: 15-29

Di Rosa M, Willoughby DA (1971) Screens for anti-inflammatory drugs. J Pharm Pharmacol 23: 297-298

Dubuisson D. Dennis SG (1977) The formalin test: a quantitative study of the analgesic effects of the morphine, meperidine and brain stem stimulation in rats and cats. Pain 4: 161-174

Eddy NB, Leimbach D (1953) Synthetic analgesics. II. Dithienylbutenyl and Dithienylbutylamines. J Pharmacol Exp Ther 107: 385-393

Ferreira MAD, Nunes ODRH, Fujimura AHY, Pessoa ODL, Lemos TLG, Viana GSB (1999) Inhibition of platelet activation by quinones isolated from Auxemma oncocalyx Taub. Res Commun Mol Pathol Pharmacol 106: 97-107

Ferreira MAD, Leal LKAM, Pessoa ODL, Lemos, TLG. Barros SBM (2001) VIANA GSB Antioxidant activity of quinones isolated from Auxemma oncocalyx Taub. Proceedings of the 6th Pharmatech; 3th Annual Meeting of the SBTF: APGI Symposium on Membrane Transport; 3th Meeting of Pharmaceutical Quality Control-ENECQ: Aug 5-8: Recife, Brazil

Figueiredo JN, Raz B, Sequin U (1998) Novel quinone methides from Salacia kraussii with in vitro antimalarial activity. J Nat Prod 61: 718-723

Fontenele JB, Sousa DC (1992) Efeitos de Hymenaea courbaril Linn., Auxemma oncocalyx Taub., Torresea cearensis Fr. All. e Jatropha gossypifolia Linn. sobre a agregacao plaquetaria. In: ENCONTRO UNIVERSITARIO DE INICIACAO A PESQUISA, 11, 1992. Fortaleza. Resumos ... Fortaleza: Universidade Federal do Ceara, p 128

Gafner S, Wolfender JL, Nianga M, Stoeckli-Evans H, Hostettmann K (1996) Antifungal and antibacterial naphthoquinones from Newboudia laevis roots. Phytochemistry 42: 1315-1320

Gardmark M, Hoglund AU, Hammarlund-Udenaes, M. (1998) Aspects of tail-flick, hot-plate and electrical stimulation tests for morphine antinociception. Pharmacol Toxicol 83: 252-258

Gong Q, Hedner T, Hedner J, Bjorkman R, Nordberg G (1991) Antinociceptive and ventilatory effects of the morphine metabolites: morphine-6-glucuronide and morphine-3-glucuronide. Eur J Pharmacol 193: 47-53

Gyires K, Torna Z (1984) The use of the writhing test in mice for screening different types of analgesics. Arch Int Pharmacodyn 267: 131-140

Hernandez-Perez M, Rabanal RM, De La Torre MC, Rodriguez B (1995) Analgesic, anti-inflammatory, antipyretic and haematological effects of Aethiopinone, an o-naphthoquinone diterpenoid from Salvia aethiopis roots and two hemisynthetic derivates. Planta Med 61: 505-509

Houghton PI, Zarka R, De Las Heras B, Hoult JR (1995) Fixed oil of Nigella sativa and derived thymoquinone inhibit eicosanoid generation in leucocytes and membrane lipid peroxidation. Planta Med 61: 33-36

Hunskaar S, Fasmer OB, Hole K (1985) Formalin test in mice, a useful technique for evaluating mild analgesics. J Neurosci Meth 14: 69-76

Ialenti A, Ianaro A, Moncada S, Di Rosa M (1992) Modulation of acute inflammation by endogenous nitric oxide. Eur J Pharmacol 211: 177-182

Itoigawa M, Kashiwada Y. Ito C. Furukawa H, Tachibana Y, Bastow KF, Lee KH (2003) Antitumor agents, 203. Carbazole alkaloid murryaquinone A and related synthetic carbazolequinones as cytotoxic agents. J Nat Prod 63 (7): 893-897

Jungler H, Sorkin LS (2000) Nociceptive and inflammatory effects of subcutaneous TNF-alfa. Pain 85: 145-151

Koster R, Anderson M, De Beer EJ (1959) Acetic acid for analgesic screening. Fed Proc 18: 412-417

Kuo SC, Chen SC, Chen LH, Wu JB, Wang JP, Teng CM (1995) Potent antiplatelet, anti-inflammatory and antiallergic isoflavanquinones from the roots of Abrus precatorius. Planta Med 61: 307-312

Lang E, Novak A, Reeh PW, Handwerker HO (1990) Chemosensitivity of fine afferents from rat skin in vitro. J Neurophysiol 63: 887-901

Leyva A, Pessoa C, Boogaerdt F, Sokaroski R, Lemos TL, Wetmore LA, Huruta RR. Moraes MO (2000) Oncocalyxones A and C, 1.4-anthracenediones from Auxemma oncocalyx: comparison with anticancer 1.9-anthracendiones. Anticancer Res 20: 1029-1031

Lino CS, Pessoa ODL, Lemos TLC, Viana GSB (1996) Estudo da atividade analgesica e antiedematogenica do extrato hidroalcoolico de Auxemma oncocalyx e oncocalyxona A. In: SIMPOSIO DE PLANTAS MEDICINAIS DO BRASIL, 14, 1996, Florianopolis. Resumos ... Florianopolis, p 95

Masso JM, Conde JR, Villar AM, Martorell J (1993) Effect of fepradinol on rat hind paw oedema induced by several inflammatory agents. J Pharm Pharmacol 45: 959-962

Morello A, Pavani M, Garbarino JA, Chamy MC, Frey C, Mancilla J (1995) Effects and mode of action of 1.4-naphthoquinones isolated from Calceolaria sessilis on tumoral cells and trypanosoma parasites. Comp Biochem Physiol Pharmacol Toxicol Endocrinol 112: 119-128

Mori K, Ushio T, Okamoto T, Kishi T, Sayo H (1998) Effect of arylthiolated 2,3-dimetoxy-1,4-benzoquinones on respiratory activity and lipid peroxidation in bovine heart mitochondria. Biol Pharm Bull 21: 293-296

Odukoya OA, Houghton PJ, Raman A (1999) Lipoxigenase inhibitors in the seeds of Aframomum danelli K. Schum. (Zingiberaceae.). Phytomedicine 6: 251-256

Parrat JR, West GB (1958) Inhibition by various substances of oedema formation in the hind-paw of the rat induced by 5-hydroxytryptamine, histamine, dextran, eggwhite and compound 48/80. Br J Pharmacol 13: 65-70

Perry NB, Blunt JW, Munro MHG (1991) A cytotoxic and antifungal 1,4-naphthoquinone and related compounds from a New Zealand brown alga, Landsburghia quercifolia. J Nat Prod 54: 978-985

Pessoa C, Mendes CS, Pessoa LO, Sabino SH, Lemos TL. Moraes MO (1992) Avaliacao da atividade antitumoral de Auxemma oncocalyx Taub (Pau Branco). In: VII REUNIAO DA FESBE, 7., Caxambu. Resumos ... Caxambu, p 168

Pessoa C, Silveira ER, Lemos TL., Wetmore LA, Moraes MO, Leyva A (2000) Antiproliferative effects of compounds derived from plants of Northeast Brazil. Phytother Res 14: 187-191

Pessoa ODL (1994) Contibuicao ao conhecimento quimico de plantas nativas do nordeste: Auxemma oncocalyx Taub. 1994. Tese (Doutorado)--Departamento de Quimica Organica, Universidade Federal do Ceara, Fortaleza

Pessoa ODL, De Lemos TLG (1997) Allantoin and fatty acid composition in Auxemma oncocalyx. Rev Bras Farm 78: 9-10

Pessoa ODL, De Lemos TLG, Silveira ER, Braz-Filho R (1993) Novel cordiachromes isolated from Auxemma oncocalyx. Nat Prod Lett 2: 145-150

Pessoa ODL, De Lemos TLG, De Carvalho MG, Braz-Filho, R. (1995) Cordiachromes from Auxemma oncocalyx. Phytochemistry 40: 1777-1786

Plone MA, Emerich DF, Lindner MD (1996) Individual differences in the hotplate test and effects of habituation on sensitivity to morphine. Pain 66: 265-270

Ronald MB, Christopher DH (1990) A bradykinin antagonist inhibits carrageenan edema in rats. Arch Pharmacol 342: 189-193

Rueff A, Dray A (1993) Pharmacological characterization of the effects of 5-hydroxytryptamine and different prostaglandins on peripheral sensory neurons in vitro. Agents Actions 38: C13-C15

Salvemini D, Wang ZQ, Wyatt PS, Bourdon DM, Marino MH, Manning PT, Currie MG (1996) Nitric oxide: a key mediator in the early and late phase of carrageenan-induced rat paw inflammation. Br J Pharmacol 118:829-838

Sauvain M, Dedet JP, Kunesch N, Poisson J, Ganter JC, Gayral P, Kunesch G (1993) In vitro and in vivo leishmanicidal activities of natural and synthetic quinoides. Phytother Res 7: 167-171

Schaible HG, Schmidt RF (1988) Excitation and sensitization of fine articular afferents from cat's knee joint by prostaglandin [E.sub.2]. J Physiol 403: 91-104

Sittie AA, Lemmich E, Olsen CE, Haviid L, Kharazmi A, Nkrumah FK, Christensen SB (1999) Structure-activity studies: in vitro antileishmanial and malarial activity of anthraquinones from Morinda lucida. Planta Med 65: 259-261

Steen KH, Steen AE, Kreysel HW, Reeh PW (1996) Inflammatory mediators potentiate pain induced by experimental tissue acidosis. Pain 66: 163-170

Teng CM, Lin CH, Lin CN, Chung MI, Huang TF (1993) Frangulin B, an antagonist of collagen-induced platelet aggregation and adhesion, isolated from Rammus formosana. Thromb Haemostas 70: 1014-1018

Tjolsen A, Berge OG, Hunskaar S, Rosland JH, Hole K (1992) The formalin test: an evaluation of the method. Pain 51: 5-17

Tripathi YB, Shukla S, Sharma M, Shukla VK (1995) Antioxidant property of Rubia cordifolia extract and its comparison with vitamin E and parabenzoquinone. Phytother Res 9: 440-443

Vazquez B, Avila G, Segura D, Escalante B (1996) Antiinflammatory activity of extracts from Aloe vera gel. J Ethnopharmacol 55: 69-75

Willis AL (1969) Parallel assay of prostaglandin-like activity in rat inflammatory exudate by means of cascade superfusion. J Pharm Pharmacol 21: 126-128

Willoughby DA, Coote E, Turk JL (1969) Complement in acute inflammation. J Pathol 97: 295-305

Winter CA, Risely EA, Nuss GW (1962) Carrageenan-induced edema in the hind paw of the rat as an assay for antiinflammatory drugs. Proc Soc Exp Biol Med 111: 544-547

M. A. D. Ferreira (1), Osmar D. R. H. Nunes (1), Juvenia B. Fontenele (3), Otilia D. L. Pessoa (2), Telma L. G. Lemos (2) and Glauce S. B. Viana (3)

(1) Departamento de Farmacia

(2) Departamento de Quimica Organica e Inorganica

(3) Departamento de Fisiologia e Farmacologia Universidade Federal do Ceara, Fortaleza--Ce--Brasil

Address

G. S. B. Viana, Departamento de Fisiologia e Farmacologia, Faculdade de Medicina, Universidade Federal do Ceara, Rua Barbosa de Freitas, 130/1100, CEP: 60170-020--Fortaleza--Ceara--Brazil

Fax: 55 (85) 288-8333; e-mail: osorio@roadnet.com.br
COPYRIGHT 2004 Urban & Fischer Verlag
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2004 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Author:Ferreira, M.A.D.; Nunes, Osmar D.R.H.; Fontenele, Juvenia B.; Pessoa, Otilia D.L.; Lemos, Telma L.G.
Publication:Phytomedicine: International Journal of Phytotherapy & Phytopharmacology
Geographic Code:1USA
Date:Apr 1, 2004
Words:6072
Previous Article:Protective role of Apigenin on the status of lipid peroxidation and antioxidant defense against hepatocarcinogenesis in Wistar albino rats.
Next Article:Evaluation of antipyretic potential of Clitoria ternatea L. extract in rats.
Topics:


Related Articles
Phytochemical analysis and analgesic properties of Curcuma zedoaria grown in Brazil.
Topical anti-inflammatory activity of Bauhinia tarapotensis leaves.
Analgesic and antiinflammatory effects of chalcones isolated from Myracrodruon urundeuva Allemao.
Extracts and constituents of Lavandula multifida with topical anti-inflammatory activity.
Antinociceptive activity and chemical composition of constituents from Caragana microphylla seeds.
Bioactivity-guided fractionation for anti-inflammatory and analgesic properties and constituents of Xanthium strumarium L.

Terms of use | Copyright © 2017 Farlex, Inc. | Feedback | For webmasters