Influence of excipients and technological process on anti-inflammatory activity of quercetin and Achyrocline satureioides (Lam.) D.C. extracts by oral route.
The flavonoids quercetin, 3-O-methylquercetin and luteolin play an important role in the anti-inflammatory activity of Achyrocline satureioides ethanol extracts when administered intraperitoneally. The present work describes the oral anti-inflammatory effect of quercetin and A. satureioides extracts and the role played by the solvent concentration, adjuvant and drying processes of freeze-drying (FD) or spray-drying (SD) on the effect. The best anti-edema effect was observed with 250 mg/kg body wt of the freeze-dried powder (FDP), prepared with 40% (v/v) ethanol (FDP40). In contrast, 250 mg/kg body wt of FDP80, prepared with ethanol 80% (ES80), did not significantly inhibit the carrageenan-induced rat paw edema. However, when ES80 was freeze-dried in the presence of polysorbate 80 (FDP80-P80) or spray-dried in the presence of colloidal silicon dioxide (CSD) and P80 (SDP80), both dried extracts became more active. Quercetin suspension in saline did not inhibit paw edema, but the mixture of quercetin with polysorbate 80 was effective in edema inhibition by the oral route. Aqueous extract (ESAQ), freeze-dried (FDPAQ, FDPAQ-P80) or spray-dried (SDPAQ) did not exhibit the edema-inhibition effect. Taken together, the results point to the following order of efficacy (at 4h, for example): FDP40 > indomethacin > SDP40 > SDP80 = FDP80-80 > Quercetin-P80. Additionally, the FDP40, SDP40 (prepared from 40% v/v ethanol added of CSD) and SDP80 reduced the total leukocyte and polymorphonuclear cell migration in the pleural cavity.
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Keywords: Achyrocline satureioides; Quercetin; Flavonoids; Anti-inflammatory activity
Infusions of influorescence of Achyrocline satureioides (Lam.) D.C., a plant known as "marcela" or "macela", are used widely in Brazilian folk medicine as a digestive, anti-spasmodic, anti-inflammatory and hypoglycemic agent, to treat gastrointestinal disorders and to reduce blood cholesterol levels (Simoes, 1984). This plant is available in Brazil from Minas Gerais to Rio Grande do Sul. Two reports on chemical composition have demonstrated that the flavonoids quercetin, 3-O-methylquercetin and luteolin are the main constituents extracted by ethanol from the influorescences (Ferraro et al., 1981; Simoes, 1984).
Pharmacological investigations of ethanol extracts from A. satureioides influorescence have indicated that these flavonoids play an important role in the anti-inflammatory activity when administered intraperitoneally (Simoes et al., 1988). However, no studies on the anti-inflammatory activity of orally administered ethanol extracts have been reported. The well-known controversy over flavonoid aglycone absorption by oral administration (Graefe et al., 1999, 2001) confirms the importance of investigating of the anti-inflammatory effect of A. satureioides ethanol extracts administered orally.
Few reports exist regarding the influence of adjuvants and technological process on the herbal extracts or herbal substances. Piskula and Terao (1998) and Azuma et al. (2002) have demonstrated the effect of lipids, emulsifiers and ethanol on the absorption of orally administered quercetin in rats. The combination of lipids and emulsifiers enhanced the absorption of quercetin significantly, demonstrating that quercetin absorption-enhancing effects on these constituents seem to be affected by quercetin solubility in the respective vehicles used for administration.
In this context, the present work was designed to investigate the anti-inflammatory activity of quercetin and A. satureioides extracts administered orally, as well as the influence of the solvent, adjuvant, freeze-drying (FD) or spray-drying (SD) processes on this activity.
Materials and methods
Influorescences from A. satureioides (Lam.) D.C. were collected in Porto Alegre, Morro Santana, Rio Grande do Sul, Brazil in March 1998. The plant was botanically identified by Marcos Sobral and a voucher specimen (ICN 9170) was deposited at the Herbarium, Departamento de Botanica (ICN), UFRGS, Porto Alegre, Brazil. The plant was dried and ground for extraction.
The following compounds were used: methanol liquid chromatography (LC) grade and phosphoric acid (Merck, Darmstadt, Germany), LC-grade water obtained in Milli-Q system (Millipore, Bedford, MA, USA), indomethacin (Sigma, St. Louis, MO, USA), carrageenan (Sigma, St. Louis, MO, USA), sodium pentobarbital (Cristalia, Sao Paulo, Brazil), polysorbate 80 (Delaware, Porto Alegre, Brazil), quercetin (Merck, Darmstadt, Germany) and colloidal silicon dioxide (CSD) (Degussa, Dusseldorf, Germany).
Preparation of A. satureioides extractive solutions (ESs)
Three ESs were prepared. An aqueous extractive solution (ESAQ) was prepared by decoction. Two other ESs, ES40 and ES80, were prepared by maceration in ethanol 40% and 80% (v/v), respectively. A plant:solvent ratio of 0.75:10 was employed for the three ESs, which were used to prepare the six dried extracts.
The percentage of extractives in ESAQ, ES40 and ES80 was determined according to the method reported by Hartke and Mutschler (1987). Samples of 20.0g of ESs were employed. Each analysis was repeated three times (Hartke and Mutschler, 1987).
Preparation of A. satureioides freeze-dried powders (FDPs)
Three A. satureioides FDPs were prepared. The first one, named FDPAQ, was obtained from 500 ml of ESAQ. The FDP40 and FDP80 were prepared, respectively, from 500 ml of ES40 and ES80 (Table 1). The extracts were freeze-dried (Edward) under the following operating conditions: temperature, -60 [degrees]C; pressure, -[10.sup.-2] bar.
Preparation of A. satureioides spray-dried powders (SDP)
An A. satureioides SDP, SDPAQ, was obtained from 500 ml of ESAQ, and the other two SDPs, SDP40 and SDP80, were prepared from 500 ml of ES40 and ES80, respectively. All the ESs were dried by atomization in a Mini-Spray Dryer Buchi 190, with two component nozzle and current flow, under the following operating conditions: inlet temperature, 157-160 [degrees]C; feed rate, 3ml/min and spraying pressure, 2 bar (Lemos-Senna et al., 1997). All the SDPs contained 50% extract and 50% CSD, as technological adjuvant.
Preparation of quercetin spray-dried powder (SDPQ)
One gram of quercetin was dispersed in a mixture of 10 ml water and added CSD. This mixture was dried under the same conditions described for SDP. The ratio quercetin:adjuvant in the SDPQ was 1:1.
LC flavonoid assay
Apparatus and chromatographic conditions
LC analysis was carried out following the method described by De Souza et al. (2002) using a Waters equipment (Milford, MA, USA): pump Waters 510, automatic controller of flow Waters 600, a Rheodyne 7125 injection valve with a 20 [micro]l loop, detector 486 UV variable-wavelength (set at 362 nm) and a Waters 746 integrator. Flavonoids were analyzed using a Shim-pack column CLC-ODS (M) RP-18, 5 [micro]m, 250 x 4[mm.sup.2] i.d. The mobile phase consisted of a mixture 53:47 (v/v) of methanol:phosphoric acid 0.16 M. The flow rate was 0.6ml/min and the analyses were carried out at 23[+ or -]1 [degrees]C.
Preparation of samples
Samples of 25 mg of the FDPs (FDPAQ, FDP40 and FDP80) were dissolved in methanol and diluted to 50.0 ml. The methanol solution was diluted in methanol-water (53:47, v/v) yielding a concentration of 75 [micro]g/ml.
Samples of 250 mg of the SDPs (SDPAQ, SDP40 and SDP80) were extracted with ethyl acetate, the solution was filtered and the ethyl acetate extract evaporated to dryness, and then made up to 25.0 ml with methanol. The methanol solution was diluted in a mixture of methanol-water (53:47, v/v) yielding a solution containing 1.5 mg/ml extracts.
The solution was filtered through a 0.45-[micro]m membrane filter (Millipore-HVHP, MA, USA). Experiments were performed in triplicate.
Male Wistar rats (180-200 g each) were purchased from the animal-breeding laboratory of Universidade Federal do Rio Grande do Sul (Porto Alegre, Brazil). The animals were left for 2 days to acclimate to room conditions. They were maintained in cages at room temperature (25[+ or -]5 [degrees]C), and fed on a standard pellet diet (Nutrilab) and water ad libitum. A minimum of eight animals was used in each group.
Carrageenan-induced rat paw edema
The anti-edema activity was evaluated using the carrageenan-induced rat paw edema test (Winter et al., 1962; Schapoval et al., 1994). The FDPs (FDPAQ, FDP40 and FDP80) and the SDPs (SDPAQ, SDP40 and SDP80) were dispersed in saline. Further, the FDPs were tested after dispersal in 1% of polysorbate 80 in saline (FDPAQ-P80, FDP40-P80 and FDP80-P80). All FDP and SDP were administered orally, at a dose of 250 mg/kg body wt, 1 h prior to the subplantar injection of carrageenan. SDP40 was also administered at a dose of 500 mg/kg body wt.
Quercetin was administered orally (20 mg/kg body wt), using saline as vehicle. A liquid dispersion of quercetin was also tested in the absence and presence of 1% polysorbate 80 (Quercetin-P80).
The control group received, by the same route, an equivalent volume of vehicle or, in cases when adjuvants were employed, equivalent volumes of the vehicle containing the respective excipients. Indomethacin (10 mg/kg body wt, oral route), dispersed in saline, was used as positive control.
Groups of eight animals each were anesthetized using a solution of sodium pentobarbital (40 mg/kg body wt, i.p.); each animal was injected with freshly prepared suspension of carrageenan (0.1 ml-0.5 mg/ml) in saline in the subplantar tissue of the left hind paw. As the control, 0.1 ml saline solution was injected into that of the right hind paw. Edema measurements were made using a plethysmometer (model 7159, Ugo Basile, Italy) prior to and 1, 2, 3 and 4 h after carrageenan injection. Mean values of treated groups were compared with mean values of a control group and analyzed statistically.
Carrageenan-induced pleurisy in rats and collection of pleural exudate
Pleurisy was induced in rats by injecting 0.1 ml of a suspension of carrageenan in saline solution (1.0 mg/ml) into the pleural cavity of the experimental animals, following Spector (1956).
The dried extracts, FDP and SDP, that presented significant inhibition of edema in the carrageenan-induced rat paw edema test were administered orally at doses of 500 and 1000 mg/kg body wt, respectively, 1 h before the inflammation was induced. After 4 h of carrageenan injection, the pleural exudate was collected, and the total leukocytes, mononuclear and polymorphonuclear were counted.
Data obtained from animal experiments were expressed as mean[+ or -]s.e.m. Statistical differences between treatment and control were tested using the Student's t-test. A value of p<0.05 was considered statistically significant.
The concentration of extractives in the ESAQ was 0.82% (w/w), 1.09% (w/w) in the ES40 and 1.25% (w/w) in the ES80. The flavonoid contents in the FDPAQ were 4.693, 0.577 and 7.280 mg/g of quercetin, luteolin and 3-O-methylquercetin (expressed in luteolin), respectively; 17.920, 3.360 and 25.027 mg/g in the FDP40; and 25.387, 3.987 and 26.187 mg/g in the FDP80. The quercetin, luteolin and 3-O-methylquerce-tin contents in the SDP SDPAQ were 2.298, 0.346 and 3.031 mg/g of powder, respectively; 2.366, 0.749 and 4.646 in the SDP40; and 3.062, 0.841 and 4.467 mg/g in the SDP80.
Table 1 shows the results of edema inhibition by FDP, quercetin and indomethacin. The indomethacin inhibited the edema in all edema measurements reaching 64.9% inhibition at the third hour. The FDPAQ and FDP80, at a dose of 250 mg/kg body wt, administered orally, did not significantly inhibit carrageenan-induced rat paw edema as compared to the control. In contrast, the FDP40, at a dose of 250 mg/kg body wt, administered orally, showed a significant response (p<0.001, Student's t-test) on edema inhibition, of 74.3%, 55.9%, 71.6% and 63.4%, respectively, at hours 1, 2, 3, and 4, more potent than indomethacin 10 mg/kg body wt (62.1%, 53.2%, 64.9% and 47.0%).
Quercetin, at a dose of 20 mg/kg body wt, administered orally, did not significantly inhibit the carrageenan-induced rat paw edema as compared to the control (Table 1).
Table 2 shows the effect of polysorbate 80 on the activity of the FDPs or quercetin in saline. The addition of polysorbate did not change the activity of the freeze-dried extract obtained from the aqueous extract solution (FDPAQ-P80) nor from the extract solution prepared with 40% ethanol (FDP40-P80). In contrast, the FDP, obtained from the extract solution prepared with 80% ethanol (FDP80-P80), became active when polysorbate was added. The FDP80-P80, instead of FDP80, inhibited edema in all measurements, presenting the highest inhibition (49.6%) at the first hour. A similar effect was observed with the Quercetin-P80, presenting the highest effect (55.3%) at the third hour (Table 2).
Table 3 shows the effect of the SD process and addition of CSD on the anti-edema action of A. satureioides SDPs. The dried extract obtained from the 40% ethanol liquid extract prepared with CSD (SDP40) reduced the edema significantly (p<0.01). The SDP80, which was obtained with CSD, reduced the edema significantly, and this effect was similar to that presented by the FDP associated with polysorbate 80 (FDP80-P80). The SDPAQ did not significantly inhibit carrageenan-induced rat paw edema as compared to the control.
Table 4 presents the results of edema inhibition by FDP40 at a dose of 250 mg/kg body wt (FDP40-250) or 500 mg/kg body wt (FDP40-500). The FDP40-250, administered by oral route, showed significant (p<0.001, Student's t-test) edema inhibition, of 74.3%, 55.9%, 71.6% and 63.4%, respectively, in the 1, 2, 3 and 4 h. The FDP40-500 inhibited the edema at the hours 2, 3 and 4, showing a significant percentage of inhibition, but a non-dose-dependent effect.
In the context of carrageenan-induced pleurisy in rats, the oral administration of FDP40, SDP40 and SDP80 significantly inhibited total leukocyte migration in the pleural cavity by, respectively, 62.4%, 53.5% and 47.9%, and reduced polymorphonuclear cell migration by 60.5%, 47.6% and 42.5%, respectively, as compared to control. The number of mononuclear cells did not change with any dried extracts (Table 5).
In the development of herbal medicines, bioguided techniques have been the methods of choice among the technological processes, especially when a group of constituents is responsible for the pharmacological activity. This approach allows optimizing formulations for the solvent selectivity, technological processing and adjuvant with a view toward maximizing the therapeutic effect.
In this study, several types of extracts, FDP and SDP, were prepared, and the influences of the solvent, drying method and excipients on anti-inflammatory activity were evaluated, with the goal of investigating the effect of orally administered A. satureioides extracts on edema. The results were compared with those of quercetin and quercetin SDP.
To determine flavonoid content in A. satureioides powders, a method based on reversed-phase LC separation combined with UV spectrometric detection was employed. FDPAQ and SDPAQ presented the lowest flavonoid concentrations. FDP80 showed a quercetin concentration slightly higher than that of FDP40; a similar profile was observed for SDP80, when compared to SDP40.
FDP40 presented the strongest anti-edema effect by oral administration at a dose of 250 mg/kg body wt, in the carrageenan-induced edema model. The significant edema inhibition observed with FDP40, in primary (1 h) and secondary (at 3 and 4 h) response, suggests its anti-edema effect occurs in the initial phase of inflammation, when the histamine, serotonin and kinins release occurs, and also later in the final phase, when prostaglandin release occurs. These results demonstrate, therefore, that FDP40 presents a complete blockade of anti-inflammatory response, more efficacious than indomethacin at 10 mg/kg body wt.
The complications involved in absorption of flavonoids administered orally (Manach et al., 1997; Graefe et al., 2001), including the poor bioavailability of quercetin aglycone in humans (Gugler et al., 1975; Graefe et al., 1999), are well known. On the other hand, its enhancement of quercetin absorption determined by lipids and emulsifiers has been demonstrated in rats (Azuma et al., 2002). The effects of A. satureioides powders could be determined by polysorbate 80 or other substances present in the extract. It is also interesting to note the fact that FDP40 and FDP80 present very different anti-inflammatory effects but not very different flavonoid concentrations. Their flavonoid concentrations of quercetin, luteolin and 3-O-methylquercetin were 17.9, 3.36 and 25.0 mg/g, respectively, for FDP40, and 25.4, 4.0 and 26.2 mg/g, respectively, for FDP80. These observations suggest that the flavonoids are not the only active constituents of the dried extracts and lend support to the hypothesis of polysaccharide contribution to the anti-inflammatory effect (Puhlmann et al., 1992). On the other hand, the comparison of the FDP40 effect with that presented by FDPAQ, the latter presenting the lowest flavonoid content, seems to confirm the Simoes statement (1998) about the important role of the flavonoids in A. satureioides anti-inflammatory activity. In this context, our hypothesis that FDP40 presents the highest efficacy due to the fact that ethanol 40% (v/v) is able to extract both flavonoids and polysaccharides, resulting in combined or synergic anti-inflammatory effects should be further investigated, without ruling out that the polysorbate effect was probably more pronounced for FDP80 because this extract contained more quercetin than the FDP40 extract.
The FDP prepared from a liquid extract containing 80% ethanol, FDP80, and the FDPs obtained from auqueous extractive solution, FDPAQ, at a dose of 250 mg/kg body wt, and quercetin at 20 mg/kg body wt, did not significantly inhibit paw edema when administered orally.
Comparing FDP40-P80, FDP80-P80 and Quercetin-P80 it seems that the extracts prepared with solvent of the higher polarity, water and ethanol (40%, v/v), did not contain active insoluble compounds in high concentration and, therefore, that the polysorbate 80 did not have a solubilizing effect. In contrast, the suspension obtained by dispersion of FDP80 and quercetin in saline after polysorbate addition (FDP80-P80 and Quercetin-P80) became active, demonstrating that a solubilizing effect probability occurred in these two cases.
SD is a method that is used to produce redispersable and soluble powders. It is employed in the present study, in addition to the adjuvant CSD to produce A. satureioides and quercetin redispersable powders. SDP80 and SDPQ presented low edema inhibition. This observation suggests that the CSD and the SD process did not promote solubilizing effects as polysorbate 80 did.
Taken together, our results demonstrate clearly the influence of polysorbate 80 on the anti-inflammatory activity of quercetin and A. satureioides extracts, administered by oral route. SD process or CSD had this effect in the powdered extract. It is important to point out that this effect was observed only for the dried extract, prepared from ES using a less polar solvent (ethanol 80% v/v), where the presence of less polar (low-water-soluble) active constituents is probable.
Carrageenan-induced pleurisy in rats is an excellent acute inflammatory model in which fluid extravasation, leukocyte migration and the various biochemical parameters involved in the inflammation response can be measured readily in the exudate (Mikami and Miyasaka, 1983). The number of total leukocyte and polymorphonuclear cells observed for the animals treated with FDP40, SDP40 and SDP80 was lower than the number of cells presented for the control groups, suggesting that these dried extracts inhibited leukocyte mobilization.
In conclusion, the A. satureioides dried powders, FDP40, SDP40, SDP80 and FDP80-P80 inhibited the acute inflammation process when administered orally. The addition of polysorbate 80 increased the anti-inflammatory effect of both quercetin and the extract prepared with a less polar solvent (FDP80), in the presence of less water-soluble substances. This finding also represents a contribution to explaining the complexity of quercetin absorption by the oral route, in which low-water solubility seems to play an important role.
The authors thank the Brazilian Government Research Agency CNPq for financial assistance, in the form of a scholarship of K.C.B. De Souza and support of the research of E.E.S. Schapoval and V.L. Bassani.
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K.C.B. De Souza, V.L. Bassani*, E.E.S. Schapoval
Programa de Pos-graduacao em Ciencias Farmaceuticas, Faculdade de Farmacia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
*Corresponding author. Tel.: +55 51 33165090; fax: +55 51 33165437.
E-mail address: email@example.com (V.L. Bassani).
Table 1. Effect of quercetin and freeze-dried powders (FDPs) from Achyrocline satureioides administered orally at a dose of 250 mg/kg body wt, on carrageenan-induced rat paw edema Edema size mean[+ or -]s.e.m. (% inhibition) Treatment 1 h 2 h Control 2.276[+ or -]0.200 2.848[+ or -]0.436 FDPAQ 2.021[+ or -]0.223 (11.2) 2.729[+ or -]0.327 (4.2) FDP40 0.586[+ or -]0.177*** (74.3) 1.255[+ or -]0.135** (55.9) FDP80 1.773[+ or -]0.155 (22.1) 2.181[+ or -]0.201 (23.4) Quercetin 1.464[+ or -]0.356 (35.7) 3.013[+ or -]0.294 (-5.8) Indomethacin 0.863[+ or -]0.264*** (62.1) 1.333[+ or -]0.213** (53.2) Edema size mean[+ or -]s.e.m. (% inhibition) Treatment 3 h 4 h Control 3.146[+ or -]0.388 3.008[+ or -]0.336 FDPAQ 3.066[+ or -]0.498 (2.5) 2.480[+ or -]0.371 (17.6) FDP40 0.894[+ or -]0.234*** (71.6) 1.101[+ or -]0.173*** (63.4) FDP80 2.490[+ or -]0.226 (20.9) 2.541[+ or -]0.264 (15.5) Quercetin 3.439[+ or -]0.287 (-9.3) 3.407[+ or -]0.255 (-13.3) Indomethacin 1.104[+ or -]0.231*** (64.9) 1.594[+ or -]0.319** (47.0) Eight animals for each experimental group; FDPAQ, freeze-dried prepared from aqueous extractive solution; FDP40, freeze-dried powder prepared from extractive solution containing 40% of ethanol; FDP80, freeze-dried powder prepared from extractive solution containing 80% of ethanol; dose = 10 mg/kg body wt for indomethacin and 20 mg/kg body wt for quercetin; control, saline. *p<0.05, **p<0.01 and ***p<0.001, compared to control group; Student's t-test. Table 2. Effect of quercetin and freeze-dried powders from Achyrocline satureioides added to 1% polysorbate 80, administered orally at a dose of 250 mg/kg body wt, on carrageenan-induced rat paw edema Edema size mean[+ or -]s.e.m. (% inhibition) Treatment 1 h 2 h Control 1.436[+ or -]0.146 2.826[+ or -]0.224 FDPAQ-P80 1.938[+ or -]0.146 (-34.9) 3.406[+ or -]0.168 (-20.5) FDP40-P80 0.713[+ or -]0.125** (50.3) 1.321[+ or -]0.364** (53.3) FDP80-P80 0.724[+ or -]0.160* (49.6) 1.518[+ or -]0.383* (46.3) Quercetin-P80 0.940[+ or -]0.136 (34.5) 1.483[+ or -]0.327* (47.5) Indomethacin 0.863[+ or -]0.264 (39.9) 1.333[+ or -]0.213** (52.8) Edema size mean[+ or -]s.e.m. (% inhibition) Treatment 3 h 4 h Control 3.604[+ or -]0.426 3.369[+ or -]0.361 FDPAQ-P80 3.939[+ or -]0.305 (-9.3) 3.536[+ or -]0.180 (5.0) FDP40-P80 1.361[+ or -]0.313*** (62.2) 1.326[+ or -]0.329** (60.6) FDP80-P80 2.109[+ or -]0.432* (41.5) 2.018[+ or -]0.452 (40.1) Quercetin-P80 1.610[+ or -]0.324** (55.3) 2.097[+ or -]0.348 (37.8) Indomethacin 1.104[+ or -]0.231*** (69.4) 1.594[+ or -]0.319** (52.7) Eight animals for each experimental group; FDPAQ-P80, freeze-dried powder aqueous added polysorbate 80; FDP40-P80, freeze-dried powder 40 added polysorbate 80; FDP80-P80, freeze-dried powder 80 added polysorbate 80; Quercetin-P80, dispersion of quercetin associated to 1% of polysorbate 80; dose = 250 mg/kg body wt for dried-powders and, 10 mg/kg body wt for indomethacin and 20 mg/kg body wt for quercetin; control, saline and polysorbate 80. *p<0.05, **p<0.01 and ***p<0.001, compared to control group; Student's t-test. Table 3. Effect of quercetin and Achyrocline satureioides spray-dried powders prepared with colloidal silicon dioxide, administered orally (500 mg/kg body wt) on carrageenan-induced rat paw edema Edema size mean[+ or -]s.e.m. (% inhibition) Treatment 1 h 2 h Control 1.450[+ or -]0.118 2.298[+ or -]0.211 SDPAQ 1.151[+ or -]0.202 (20.6) 2.394[+ or -]0.512 (-4.2) SDP40 0.849[+ or -]0.095** (41.4) 0.986[+ or -]0.306** (57.1) SDP80 0.749[+ or -]0.199** (48.3) 1.484[+ or -]0.169* (35.4) Indomethacin 0.863[+ or -]0.264* (40.5) 1.333[+ or -]0.213** (42.0) Edema size mean[+ or -]s.e.m. (% inhibition) Treatment 3 h 4 h Control 2.375[+ or -]0.250 2.612[+ or -]0.286 SDPAQ 2.855[+ or -]0.494 (-20.2) 2.340[+ or -]0.506 (10.4) SDP40 0.991[+ or -]0.413** (58.3) 1.149[+ or -]0.343** (56.0) SDP80 1.546[+ or -]0.182* (34.9) 1.563[+ or -]0.249* (40.2) Indomethacin 1.104[+ or -]0.231** (53.5) 1.594[+ or -]0.319* (39.0) Eight animals for each experimental group; SDPAQ, spray-dried powder obtained from aqueous extractive solution; SDP40, spray-dried powder prepared from extractive solution containing ethanol 40%; SDP80, spray- dried powder prepared from extractive solution containing ethanol 80%; dose = 10 mg/kg body wt for indomethacin; control, saline and colloidal silicon dioxide. *p<0.05, **p<0.01 and ***p<0.001, compared to control group; Student's t-test. Table 4. Effect of freeze-dried powders of Achyrocline satureioides prepared from extract solution containing 40% ethanol, administered orally at doses of 250 (FDP40-250) and 500 (FDP40-500) mg/kg body wt, on carrageenan-induced rat paw edema Edema size mean[+ or -]s.e.m. (% inhibition) Treatment 1 h 2 h Control 2.276[+ or -]0.200 2.848[+ or -]0.436 FDP40-250 0.586[+ or -]0.177*** (74.3) 1.255[+ or -]0.135** (55.9) FDP40-500 0.906[+ or -]0.256** (60.2) 1.158[+ or -]0.413** (59.3) Indomethacin 0.863[+ or -]0.264*** (62.1) 1.333[+ or -]0.213** (53.2) Edema size mean[+ or -]s.e.m. (% inhibition) Treatment 3 h 4 h Control 3.146[+ or -]0.388 3.008[+ or -]0.336 FDP40-250 0.894[+ or -]0.234*** (71.6) 1.101[+ or -]0.173*** (63.4) FDP40-500 1.194[+ or -]0.443** (62.0) 0.958[+ or -]0.532*** (68.2) Indomethacin 1.104[+ or -]0.231*** (64.9) 1.594[+ or -]0.319** (47.0) Eight animals for experimental group FDP-250 and six animals for experimental group FDP-500; dose = 10 mg/kg body wt for indomethacin; control, saline. *p<0.05, **p<0.01 and ***p<0.001, compared to control group; Student's t-test. Table 5. Leukocyte migration to pleural cavity after 4 h of the inflammation induction in rats treated with dried powders obtained from the Achyrocline satureioides by oral route, and of the control group (saline) Mononuclear TLC cells x cells x Treatment [10.sup.5][+ or -]s.e.m. [10.sup.5][+ or -]s.e.m. FDP40 (500 mg/kg 75.1[+ or -]8.4** 17.2[+ or -]2.1 body wt) Control 120.3[+ or -]9.1 24.6[+ or -]3.5 SDP40 (1000 mg/kg 38.8[+ or -]3.5* 12.8[+ or -]1.2 body wt) Control 72.6[+ or -]12.9 17.8[+ or -]2.7 SDP80 (1000 mg/kg 34.8[+ or -]6.8* 11.5[+ or -]2.3 body wt) Control 72.6[+ or -]12.9 17.8[+ or -]2.7 Polymorphonuclear Treatment cells x [10.sup.5][+ or -]s.e.m. FDP40 (500 mg/kg 57.9[+ or -]7.3** body wt) Control 95.7[+ or -]9.1 SDP40 (1000 mg/kg 26.1[+ or -]3.6* body wt) Control 54.8[+ or -]10.4 SDP80 (1000 mg/kg 23.3[+ or -]5.0** body wt) Control 54.8[+ or -]10.4 FDP40, freeze-dried powder prepared from extractive solution containing 40% of ethanol; SDP40, spray-dried powders prepared from extractive solution containing ethanol 40% with colloidal silicon dioxide; SDP80, spray-dried powders prepared from extractive solution containing ethanol 80% with colloidal silicon dioxide; TLC, total leukocyte count. *p<0.05 and **p<0.01, compared to control rats; Student's t-test.
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|Author:||De Souza, K.C.B.; Bassani, V.L.; Schapoval, E.E.S.|
|Publication:||Phytomedicine: International Journal of Phytotherapy & Phytopharmacology|
|Date:||Feb 1, 2007|
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