Valuation of anti-inflammatory and antinociceptive activities of Erica species native to Turkey

Erica L. species (Ericaceae) have been popularly used as antirheumatic, diuretic, astringent and treatment of urinary infections. In order to evaluate this information, anti-inflammatory and antinociceptive activities of different extracts prepared with methanol, chloroform, ethyl acetate, n-butanol and water from the aerial parts of Erica arborea L., Erica manipuliflora Salisb., Erica bocquetii (Pe?men) P.F. Stevens and Erica sicula Guss. subsp. libanotica (C.&;W. Barbey) P.F. Stevens (Ericaceae) of Turkish origin were investigated by using in vivo methods. For the anti-inflammatory activity, carrageenan-induced hind paw edema model, PGE₂-induced hind paw edema model, and 12-O-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema model and for the antinociceptive activity p-benzoquinone-induced writhing test in mice were employed. The ethyl acetate extracts of Erica arborea (EAE), Erica bocquetii (EBE) and Erica manipuliflora (EME) exhibited notable inhibition against carrageenan-induced (24.1–32.3%, 23.8–36.1%, 29.2–35.1%, respectively) and PGE₂-induced (21.2–37.7%, 6.8–29.7%, and 6.2–34.1%, respectively) hind paw edema as well as TPA-induced mouse ear edema models in mice, while the ethyl acetate extract of Erica sicula subsp. libanotica (ESE) (10.7–29.7%) displayed potent anti-inflammatory activity only on the PGE₂-induced hind paw edema model. However, the remaining extracts were found to be inactive against inflammatory models. Same extracts, i.e., EAE, EBE and EME were also found to exhibit remarkable antinociceptive activity in p-benzoquinone-induced abdominal constriction test at a dose of 100 mg/kg (46.5%, 27.7% and 36.3%, respectively).     

Assessment of anti-inflammatory and antinociceptive activities of Daphne pontica L. (Thymelaeaceae)

The n-hexane, diethylether, ethyl acetate and methanol extracts from roots, leaves, stems and flowers with young leaves of Daphne pontica L. (Thymelaeaceae) were investigated for their in vivo anti-inflammatory and antinociceptive activities. For the anti-inflammatory activity assessment, carrageenan-induced hind paw edema, PGE(2)-induced hind paw edema and 12-O-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema models and for the antinociceptive activity, p-benzoquinone-induced abdominal constriction test were used. Only ethyl acetate extracts of the roots showed significant anti-inflammatory activity on carrageenan-induced (22.7-32.0% inhibition) and PGE(2)-induced hind paw edema (3.2-27.3% inhibition) as well as 12-O-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema (47.8-43.3% inhibition) models at 50 mg/kg dose without inducing any apparent gastric lesion or acute toxicity, whereas the other extracts were shown to be ineffective. In addition to roots, ethyl acetate extracts of the stems exhibited 19.5-29.9%; 5.3-23.9%; 36.6-28.1% inhibition on carrageenan-induced and PGE(2)-induced hind paw edema as well as 12-O-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema models, respectively. On the other hand, none of the extracts showed any significant antinociceptive activity.     

Estimation of antinociceptive and anti-inflammatory activity on Geranium pratense subsp. finitimum and its phenolic compounds

To obtain experimental evidence on the therapeutic efficacy of Geranium species and its phenolic compounds for inflammatory diseases, we examined the effects of the aqueous extract of the aerial parts of Geranium pratense subsp. finitimum (Woronow) Knuth, its fractions and isolated compounds, the mixture of quercetin 3-O-alpha-arabinopyranoside, kaempferol 3-O-beta-galactopyranoside (1), the mixture of quercetin 3-O-beta-glucopyranoside, quercetin 3-O-beta-galactopyranoside (2), kaempferol 3-O-beta-glucopyranoside (3), (-)-6-chloroepicatechin (4), the mixture of quercetin 3-O-(2'-O-galloyl)-beta-glucopyranoside, quercetin 3-O-(2'-O-galloyl)-beta-galactopyranoside (5) and myricetin 3-O-(2'-O-galloyl)-beta-glucopyranoside (6), on carrageenan-, PGE(2)- and TPA-induced inflammation in mice and p-benzoquinone-induced writhing reflex to assess anti-inflammatory and antinociceptive activities. The effective dose of materials for the inhibition of carragenan-induced hind paw edema assay was determined to be 100 mg/kg, which was also used in the assays with the extract, its fractions and isolated compounds in all other experiments. The aqueous extract, 1, 2 (100 mg/kg), as well as indomethacin (10 mg/kg) inhibited significantly the formation of the carrageenan-induced hind paw edema. There was also a significant reduction in PGE(2)-induced hind paw edema and TPA-induced ear edema models with 5, in addition to the aqueous extract and the other active components 1 and 2. In the antinociceptive assay, the aqueous extract and its fractions, as well as 1, 2, and 5 diminished significantly the number of writhings. Based on the results obtained it is suggested that the aqueous extract of Geranium pratense subsp. finitimum and its phenolic compounds display anti-inflammatory activity, supporting the folkloric use.     

In vivo anti-inflammatory and antinociceptive activity of the crude extract and fractions from Rosa canina L. fruits

The aqueous and ethanol extracts of Rosa canina L. (Rosaceae) fruits and the fractions prepared from the latter were investigated for their anti-inflammatory and antinociceptive activities in several in vivo experimental models. The ethanolic extract was shown to possess significant inhibitory activity against inflammatory models (i.e., carrageenan-induced and PGE(1)-induced hind paw edema models, as well as on acetic acid-induced increase in a capillary permeability model) and on a pain model based on the inhibition of p-benzoquinone-induced writhing in mice. Hexane, chloroform, ethylacetate, n-butanol and the remaining water fractions were obtained through bioassay-guided fractionation. Ethylacetate and n-butanol fractions displayed potent anti-inflammatory and antinociceptive activities at a dose of 919 mg/kg without inducing acute toxicity. Further attempts to isolate and define the active constituent(s) were inconclusive, possibly due to the synergistic interaction of components in the extract.     

Anti-inflammatory and antinociceptive potential of Maclura pomifera (Rafin.) Schneider fruit extracts and its major isoflavonoids, scandenone and auriculasin

The aqueous, ethanolic and chloroform extracts and two prenylated isoflavones: scandenone (I) and auriculasin (II), isolated from the fruits of Maclura pomifera (Rafin.) Schneider, were investigated for their in vivo anti-inflammatory and antinociceptive activity. For the anti-inflammatory activity, both carrageenan-induced hind paw edema and 12-O-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema models and for the antinociceptive activity, p-benzoquinone-induced abdominal constriction test were used. Scandenone, the chloroform and the ethanolic extracts were shown to possess antinociceptive activity and anti-inflammatory activity on carrageenan-induced hind paw edema model at 100 mg/kg dose. The same compound and the extract were also found to be highly active in (TPA)-induced mouse ear edema model whereas auriculasin and the H(2)O extract showed to be inactive in all of the assays.     

A comparative study on the antinociceptive and anti-inflammatory activities of five Juniperus taxa

Ethnopharmacological relevance

Juniperus L. (Cupressaceae) species have been used to various inflammatory and infectious diseases such as bronchitis, colds, cough, fungal infections, hemorrhoids, gynecological diseases, and wounds in Turkish folk medicine.

Aim of the study

To evaluate this traditional information, anti-inflammatory and antinociceptive activities of the methanolic and aqueous extracts prepared from different parts (stem, fruit and leaves) of the five Turkish taxa under Juniperus section of the gender; J. drupacea, J. communis var. communis, J. communis var. saxatilis, J. oxycedrus subsp. oxycedrus, and J. oxycedrus subsp. macrocarpa growing were investigated.

Methods

For the anti-inflammatory activity, carrageenan-induced and PGE₂-induced hind paw edema models, and for the antinociceptive activity p-benzoquinone-induced writhing and hot plate tests in mice were employed.

Results

The methanolic extracts of fruit and leaves from J. oxycedrus subsp. oxycedrus and J. communis var. saxatilis exhibited notable inhibition in carrageenan-induced edema model at a dose of 100 mg/kg. The same extracts also displayed significant activity against PGE₂-induced edema model. While, the remaining extracts were found inactive against these edema models. A similar activity pattern was observed against p-benzoquinone-induced abdominal constriction test without inducing any gastric damage or apparent acute toxicity, whereas all extracts were inactive in hot plate test.

Conclusion

The experimental data demonstrated that J. oxycedrus subsp. oxycedrus and J. communis var. saxatilis displayed remarkable anti-inflammatory and antinociceptive activities; however, further studies are warranted to define and isolate the active anti-inflammatory and antinociceptive components from these active species which may yield safe and effective agents to be used in the treatment of inflammatory disorders.     

Anti-inflammatory and antinociceptive activities of Seseli L. species (Apiaceae) growing in Turkey

The ethyl acetate and methanol (80%) extracts obtained from 10 Seseli L. species (Apiaceae) growing in Turkey, Seseli andronakii Woron., Seseli campestre Besser, Seseli gummiferum Pall. ex Sm. subsp. corymbosum (Boiss. and Heldr.) P.H. Davis, Seseli gummiferum Pall. ex Sm. subsp. gummiferum, Seseli hartvigii Parolly and Nordt, Seseli libanotis (L.) W. Koch, Seseli petraeum M. Bieb., Seseli peucedanoides (Bieb.) Koso-Pol., Seseli resinosum Freyn and Sint., Seseli tortuosum L. were evaluated for their in vivo anti-inflammatory and antinociceptive activities. For the preliminary screening of anti-inflammatory activity, carrageenan-induced hind paw edema and for the antinociceptive activity, p-benzoquinone-induced abdominal constriction test were used. Among the plant extracts, the ethyl acetate extracts from Seseli andronakii, Seseli campestre, Seseli gummiferum subsp. corymbosum, Seseli petraeum, Seseli resinosum and Seseli tortuosum showed 30.1, 32.3, 36.9, 39.8, 35.1, 37.6% inhibition in p-benzoquinone-induced abdominal constriction test, respectively. The ethyl acetate extracts of Seseli gummiferum subsp. corymbosum, Seseli petraeum, Seseli resinosum and Seseli tortuosum also exhibited notable inhibition, ranging between 24.5-29.7, 28.1-33.3, 17.4-27.5 and 27.9-31.3%, respectively, in carrageenan-induced hind paw edema model at 100 mg/kg dose without inducing any gastric damage, quite comparable to indomethacin (41.8-44.8% inhibition) as a reference sample. During the acute toxicity evaluation, neither death nor gastric bleeding was observed for any of the plant extracts. Results have supported the traditional use of some Seseli species against inflammatory disorders. Further studies are warranted to define and isolate the active anti-inflammatory and antinociceptive components from the active species which may yield safe and effective agents to be used in current treatments.     

Evaluation of anti-inflammatory and antinociceptive activities of some Onosma L. species growing in Turkey

ETHNOPHARMACOLOGICAL RELEVANCE: Roots of Onosma species are used for the treatment of various disorders such as bronchitis, tonsillitis, hemorrhoids as well as alleviating pains in folk medicine in Turkey. AIM OF THE STUDY: The chloroform and ethanol (70%) extracts obtained from the roots of Onosma species (Boraginaceae) growing in Turkey, Onosma aucheranum DC., Onosma isauricum Boiss. and Heldr. (endemic), Onosma sericeum Willd., Onosma tauricum Pallas ex Willd. var. brevifolium DC. (endemic) and Onosma tauricum Pallas ex Willd. var. tauricum (Syn: Onosma velenovskyi Davidov) were evaluated for in vivo anti-inflammatory and antinociceptive activities. MATERIALS AND METHODS: For the preliminary screening, carrageenan-induced hind paw edema for the anti-inflammatory activity and, p-benzoquinone-induced abdominal constriction test for the antinociceptive activity were used in mice. RESULTS: The chloroform extracts from Onosma aucheranum and Onosma isauricum and ethanolic extracts from Onosma isauricum and Onosma sericeum demonstrated 28.0%, 34.3%, 24.6% and, 27.5% inhibition in p-benzoquinone-induced abdominal constriction test. The chloroform and ethanol (70%) extracts of Onosma isauricum and ethanol (70%) extract of Onosma sericeum also exhibited marked inhibition, ranging between 12.3-27.3%, 10.5-25.3%, 8.2-22.6%, respectively, in carrageenan-induced hind paw edema model at 100mg/kg dose without gastric damage and the activity was quite comparable to indomethacin (32.0-38.4% inhibition) as a reference sample. Neither death nor gastric bleeding was observed for any of the plant extracts during the acute toxicity evaluation. CONCLUSION: The experimental data demonstrated that Onosma aucheranum, Onosma isauricum and Onosma sericeum displayed remarkable anti-inflammatory and antinociceptive activities.     

Biological activities of Pseudevernia furfuracea (L.) Zopf extracts and isolation of the active compounds

Ethnopharmacological relevance

Pseudevernia furfuracea (L.) Zopf (Parmeliaceae) is a common epiphytic lichen in the conifer-hardwood forest of Anatolia. This species is used in traditional medicine in Turkey as a treatment for wounds, eczema and hemorrhoids.

Aim of the study

The present study was designed to investigate the active compounds from Pseudevernia. furfuracea, and the isolation studies yielded atraric acid ( Aslan et al., 2006) as the major compound and a mixture of methyl hematommate ( Baumann, 1960) and methyl chlorohematommate ( Bay?r et al., 2006). Furthermore, methanolic extract from thalli of Pseudevernia. furfuracea and its fractions and isolates ( 1, 2 and 3) were investigated for in vitro antimicrobial and antioxidant activities, and in vivo antinociceptive, anti-inflammatory and wound healing activities.

Material and methods

Antimicrobial activities of the samples were determined by using the disc diffusion technique. 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was used as a rapid TLC screening method to evaluate the antioxidant activity of Pseudevernia. furfuracea. The thiobarbituric acid (TBA) test was used to assess the efficacy of the extracts in protecting liposomes from lipid peroxidation. In vivo inhibitory effect of the extracts on the carrageenan-induced hind paw edema model in mice was studied for the assessment of anti-inflammatory activity. p-Benzoquinone-induced abdominal constriction test was used to explore the antinociceptive effects of the extracts. Moreover, the wound healing potential of the plant extracts that were evaluated by using in vivo incision and excision wound models on rats and mice, were comparatively assessed with a reference ointment Madecassol^®.

Results

Significant antimicrobial activities were observed against Gram (+) microorganisms and Candida krusei and Candida. dubliniensis in dichloromethane (DCM) and ethyl acetate (EtOAc) extracts and isolates. The methanol (MeOH), DCM and EtOAc extracts of the lichen were found to possess moderate inhibitory activity on lipid peroxidation. Methanolic extract of the lichen was found to possess significant inhibitory activity on the carrageenan-induced hind paw edema model in mice whereas the other fractions did not show any activity. While DCM and EtOAc extracts and fractions showed notable anti-inflammatory activity on carrageenan-induced hind paw edema model without inducing any apparent acute toxicity or gastric damage. Moreover, topical application of the ointment prepared with MeOH extract and EtOAc fraction onto the incised wounds exerted remarkable wound healing activity.

Conclusion

The results of these experimental studies exhibited that nonpolar fractions of Pseudevernia. furfuracea have significant antimicrobial activity against especially Candida species and polar fractions (especially MeOH) display antioxidant, anti-inflammatory, antinociceptive and wound healing activities.     

Flavonoids with anti-inflammatory and antinociceptive activity from Cistus laurifolius L. leaves through bioassay-guided procedures

Extracts obtained from the leaves and branches of various Cistus species have been used worldwide as folk remedy for the treatment of various inflammatory ailments including rheumatism and renal inflammations. Effects of the extracts and fractions from the leaves with non-woody branches of Cistus laurifolius L. (Cistaceae) were studied using two in vivo models of inflammation in mice. Model one was based on observed potent inhibitory activity against carrageenan-induced hind paw oedema and the second model used was acetic acid-induced, increased vascular permeability model. Through bioassay-guided fractionation and isolation procedures three flavonoids; 3-O-methylquercetin (1), 3,7-O-dimethylquercetin (2) and 3,7-O-dimethylkaempferol (3) were isolated as the main active ingredients from the ethanol extract. Later on, these flavonoids were shown to possess potent antinociceptive activity, which was assessed through inhibition of p-benzoquinone-induced writhing reflex. Results of the present study have clearly supported the utilization of Cistus laurifolius in Turkish traditional medicine. Subsequently, three flavonoids were shown to have strong antinociceptive and anti-inflammatory activities, per os without inducing any apparent acute toxicity as well as gastric damage.     

Anti-inflammatory and antinociceptive activity assessment of plants used as remedy in Turkish folk medicine

Ethanolic and aqueous extracts from seven plant species used in Turkish traditional medicine were evaluated for in vivo anti-inflammatory and antinociceptive activities; Helleborus orientalis Lam. roots and herbs, Juglans regia L. leaves, Laurocerasus officinalis Roemer leaves, Nerium oleander L. dried and fresh flowers and leaves, Rhododendron ponticum L. leaves, Rubus hirtus Walds. et Kit aerial parts and Rubus sanctus Schreber aerial parts and roots. All the plant extracts, except the aqueous extract of Rubus hirtus, were shown to possess significant antinociceptive activity in varying degrees against p-benzoquinone-induced abdominal contractions in mice. However, only the ethanolic extracts of Helleborus orientalis roots, Juglans regia leaves, Laurocerasus officinalis leaves, Nerium oleander dried and fresh flowers, and Rhododendron ponticum leaves exhibited potent anti-inflammatory activity against carrageenan-induced hind paw edema model in mice without inducing any gastric damage. Results of the present study confirmed the folkloric claim that all the selected materials to possess potent antinociceptive and anti-inflammatory activity.     

Ethnopharmacological evaluation of some Scorzonera species: in vivo anti-inflammatory and antinociceptive effects

Ethnopharmacological relevance

Scorzonera species are mainly used against inflammation and to relieve pain in Turkish traditional medicine. Therefore, we aimed to assess in vivo anti-inflammatory and antinociceptive activities of the aerial part and root extracts of Scorzonera acuminata, Scorzonera cana var. alpina, Scorzonera cana var. jacquiniana, Scorzonera cana var. radicosa, Scorzonera cinerea, Scorzonera eriophora, Scorzonera incisa, Scorzonera laciniata ssp. laciniata, Scorzonera parviflora and Scorzonera sublanata.

Materials and methods

For the anti-inflammatory activity evaluation carrageenan, PGE₂ and serotonin-induced hind paw edema and 12-O-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema models were used. p-Benzoquinone-induced abdominal constriction test was employed in mice for the assessment of antinociceptive activity. Furthermore chemical composition of the tested extracts was investigated qualitatively and quantitatively by using RP-HPLC method. Some phenolic acids and flavonoids were used as standards.

Results

Extracts prepared from the aerial parts of Scorzonera cana var. jacquiniana, Scorzonera cinerea, Scorzonera eriophora, Scorzonera incisa and Scorzonera parviflora showed significant inhibitory effect on carrageenan and PGE₂-induced hind paw edema model as well as on p-benzoquinone-induced abdominal constriction test. The extracts did not show any remarkable activity on serotonin-induced hind paw edema and TPA-induced mouse ear edema models. Chlorogenic acid was detected as major compounds in all the species investigated. Additionally, among the tested flavonoids, luteolin-7-glucoside, hyperoside and rutin were found to be in different amounts in Scorzonera species.

Conclusion

The experimental data revealed that Scorzonera cana var. jacquiniana, Scorzonera cinerea, Scorzonera eriophora, Scorzonera incisa and Scorzonera parviflora possess significant anti-inflammatory and antinociceptive activity. It has been suggested that flavonoids and chlorogenic acid are partly responsible for mentioned activities of Scorzonera species.     

Estimation of anti-inflammatory,antinociceptive and antioxidant activities on Arctium minus (Hill) Bernh. ssp. minus

Ethnopharmacological relevance

Arctium minus (Hill) Bernh. ssp. minus (Asteraceae) leaves are used to alleviate rheumatic pain, against fever and sunstroke with externally application in Turkish folk medicine.

Aim of the study

To evaluate the anti-inflammatory, antinociceptive and antioxidant activities of aqueous and ethanol extracts prepared from the leaves of Arctium minus ssp. minus.

Materials and methods

The ethanolic and aqueous extracts from the leaves of Arctium minus ssp. minus were evaluated in mice for anti-inflammatory activity using carrageenan-induced hind paw edema model and for antinociceptive activity using p-benzoquinone-induced abdominal contractions test. Moreover, the antioxidant power of the extracts has been determined by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and flow injection analysis-luminol chemiluminescence (FIA-CL). In addition, the total phenolic content in both extracts was determined with spectrophotometric method.

Results

Our results showed that only the ethanol extract exhibited a dose-dependent anti-inflammatory activity ranging between 11.1 and 23.6% at 200 mg/kg dose as well as displayed a significant antinociceptive activity without inducing any gastric damage. Although, both extracts were shown to possess significant DPPH radical-scavenging activity, that of aqueous extract was found to have more pronounced activity. In FIA-CL system, the ethanol extract was shown to possess a significant scavenger activity against H₂O₂ while the aqueous extract was much more potent antioxidant activity against HOCl-luminol CL than ethanol extract.

Conclusion

According to our results, it was concluded that Arctium minus ssp. minus contains potent natural antioxidants. In this study, in vivo experimental results have also supported the folk medicinal utilization of Arctium minus ssp. minus.     

Bioactivity guided evaluation of anti-inflammatory and antinociceptive activities of Arceuthobium oxycedri (D.C.) M. Bieb.

Ethnopharmacological relevance

Arceuthobium oxycedri (D.C.) M. Bieb. (Loranthaceae) or dwarf mistletoe is a rare semi-parasitic plant that lives on the branches of Juniperus oxycedrus as the host plant. In Turkish folk medicine, the plant is prescribed as a panacea for every kind of diseases, including, against infectious and inflammatory disorders of upper respiratory system and gastro-intestinal complaints or as a hypotensive remedy.

Aim of the study

In the current study, in vivo anti-inflammatory and antinociceptive effects of Arceuthobium oxycedri have been investigated.

Material and methods

The crude ethanolic extract of the whole plant was sequentially fractionated into five subextracts; explicitly, n-hexane, chloroform, ethyl acetate (EtOAc), n-butanol, and remaining water extracts. Further studies were carried out on the most active subextract, i.e. the EtOAc subextract, was further subjected to fractionation through successive column chromatographic applications on Silica gel 60, Sephadex LH-20 and LiChropep RP-18. For the activity assessment, each extract or fraction was submitted to bioassay systems; carrageenan-induced hind paw edema model for anti-inflammatory activity and p-benzoquinone induced abdominal contraction test for antinociceptive activity assessment.

Results

Among the extracts obtained, the ethanolic extract, EtOAc and n-butanol subextracts showed significant inhibitory activity in the bioassay systems. From the EtOAc subextract, a major component was isolated and its structure was determined as (+)-catechin by means of spectral techniques.

Conclusion

Present study confirms the claimed utilization of the plant against inflammatory complaints in Turkish folk medicine.     

Anti-inflammatory and antinociceptive effects of the stem bark of Byrsonima intermedia A. Juss

Ethnopharmacological relevance

Byrsonima intermedia A. Juss. is popularly known as “murici pequeno” and is native to the Brazilian Cerrado. This species has been used as an antimicrobial, anti-hemorrhagic, anti-diarrheal and anti-inflammatory. Nevertheless, scientific information regarding Byrsonima intermedia is limited; there are no reports related to its possible anti inflammatory and antinociceptive effects. This study employed in vivo inflammatory and nociceptive models to evaluate the scientific basis for the traditional use of Byrsonima intermedia.

Materials and methods

Carrageenan-induced paw edema, peritonitis and fibrovascular tissue growth induced by s.c. cotton pellet implantation tests were used to investigate the anti-inflammatory activity of Byrsonima intermedia aqueous extract (BiAE) in rats. Mechanical nociceptive paw, formalin and hot plate tests were used to evaluate the antinociceptive activity in mice. High-performance liquid chromatography (HPLC), phytochemistry screening and determination of total phenolics and flavonoids were used to determine the chemical profile of the BiAE.

Results

BiAE at test doses of 30-300 mg/kg p.o. clearly demonstrated anti-inflammatory effects by reduced carrageenan-induced paw edema, by inhibited leukocyte recruitment into the peritoneal cavity and, in the model of chronic inflammation, by using the cotton pellet-induced fibrovascular tissue growth in rats. The extracts at test doses of 30-300 mg/kg p.o. clearly demonstrated antinociceptive activity in all tests. Administration of the opioid receptor antagonist naloxone completely inhibited the antinociceptive effect induced by BiAE (100 mg/kg).

Conclusion

BiAE markedly exhibits anti-inflammatory action in rats and antinociceptive activity in mice. Thus, it may be useful in the treatment of inflammatory hyperalgesic disorders, which supports previous claims of its traditional use.     

Molecular mechanism of anti-inflammatory activity of Pluchea indica leaves in macrophages RAW 264.7 and its action in animal models ofinflammation

Ethnopharmacological relevance: Pluchea indica Less.

(Asteraceae) is a Thai medicinal plant used in traditional medicine for the treatment of hemorrhoids, lumbago, leucorrhoea and inflammation. This study investigated the molecular mechanism of anti-inflammatory activity of Pluchea indica leaf extract in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and also determined its action in acute inflammation animal models.

Materials and methods

The inhibitory effect of Pluchea indica leaf extract on LPS-induced nitric oxide (NO) production was evaluated by Griess reaction. Protein and mRNA expressions were determined by real time RT-PCR and Western blotting analysis, respectively. Inducible nitric oxide synthase (iNOS) promoter activity was evaluated by iNOS promoter based reporter gene assay. In vivo anti-inflammatory effect was examined in ethylphenylpropiolate (EPP)-induced ear edema and carrageenan-induced paw edema in rat models.

Results

Ethyl acetate fraction of ethanol extract of Pluchea indica leaves (EFPI) exhibited the potent inhibitory effect on NO production in LPS-induced macrophages and also inhibited PGE₂ release. EFPI reduced iNOS mRNA and protein expression through suppressed iNOS promoter activity and nuclear translocation of subunit p65 of nuclear factor-κB, but did not inhibit phosphorylation of the mitogen-activated protein kinases (MAPKs). Moreover, EFPI possessed anti-inflammatory activities on acute phase of inflammation as seen in EPP-induced ear edema and carrageenan-induced paw edema inrats.

Conclusions

These data support the pharmacological basis of Pluchea indica plant as a traditional herbal medicine for treatment of inflammation.     

Flavonoids with antinociceptive and anti-inflammatory activities from the leaves of Tilia argentea (silver linden)

Silver linden, Tilia argentea Desf. ex DC (Tiliaceae) leaves are used in the treatment of common cold and bronchitis. In order to evaluate this information, antinociceptive and anti-inflammatory activities of the two main flavonoid glycosides: kaempferol-3,7-O-alpha-dirhamnoside and quercetin-3,7-O-alpha-dirhamnoside isolated from the leaves, were investigated. For the antinociceptive activity, p-benzoquinone-induced writhing test and for the anti-inflammatory activity, carrageenan-induced hind paw edema model in mice were used. Both compounds were shown to possess potent antinociceptive and anti-inflammatory activity at 50 mg/kg dose, per os, without inducing any apparent acute toxicity as well as gastric damage.     

Experimental study on anti-inflammatory activity of a TCM recipe consisting of the supercritical fluid CO2 extract of Chrysanthemum indicum, Patchouli Oil and Zedoary Turmeric Oil in vivo

Ethnopharmacological relevance

Chrysanthemum indicum (Compositae) Linné, Pogostemon cablin (Blanco) Benth and Curcuma wenyujin (Zingiberaceae) Y. H. Chen et C. Ling are three of the extensively used herbal remedies among traditional Chinese medicines for the purpose of anti-inflammation. A traditional Chinese medicine (TCM) recipe named CPZ consisting extracts of the above three herbs, has shown noteworthy anti-influenza activity, which is closely related to its anti-inflammatory feature.

Aim of this study

To investigated the anti-inflammtory activity of CPZ in vivo for a further exploration of the recipe's anti-inflammatory properties.

Materials and methods

The anti-inflammatory property of CPZ on acute inflammation was evaluated by inflammatory models of dimethylbenzene (DMB)-induced ear vasodilatation and acetic acid-induced capillary permeability enhancement in mice, as well as the carrageenan-induced paw edema rat model, in which inflammation-related cytokine including prostaglandin E₂ (PGE₂), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) in the edematous paw tissue were determined by enzyme linked immunosorbent assay (ELISA). Moreover, effect of CPZ on chronic inflammation was observed through granuloma formation in rats subjected to cotton pellet implantation.

Results

CPZ (340, 170, and 85 mg/kg for mice, p.o.) not only decreased the DMB-induced ear vasodilatation but also attenuated capillary permeability under acetic acid challenge in mice. And the significant inhibition on carrageenan-induced paw edema was observed. Further more, the ELISA results showed that CPZ (170, 85, and 42.5 mg/kg for rats, p.o.) could up-regulate the level of IL-1β in the edema paw tissue of rats significantly while down-regulate that of PGE₂, but no apparent effect on TNF-α or NO was observed in the test. Besides, CPZ had a certain degree of restraining effect on the cotton pellet-induced granuloma formation in rats and the highest dose of 170 mg/kg even showed a significant suppression on it.

Conclusion

The above results indicated that CPZ possessed a potent anti-inflammatory activity, which is indicated to be closely associated with its regulation on IL-1β and PGE₂ thereby mediating the inflammatory response acting at an appropriate level.     

Inhibitory effect of eupatilin and jaceosidin isolated from Artemisia princeps on carrageenan-induced inflammation in mice

Ethnopharmacological relevance

Artemisia princeps Pampanini (family Asteraceae) is an herbal medicine widely used as a hepatoprotective, antioxidative, anti-inflammatory, and antibacterial agent in Korea, China, and Japan.

Aim of the study

This study aimed to elucidate the anti-inflammatory effect of the main constituents, eupatilin and jaceosidin, isolated from Artemisia princeps.

Materials and methods

We used carrageenan-induced inflammation in an air pouch on the back of mice and carrageenan-induced hind paw edema in rats to determine the anti-inflammatory effects of eupatilin and jaceosidin. Inflammatory makers, such as expression of pro-inflammatory cytokines and cyclooxygenase (COX)-2, and activation of nuclear factor-kappa B (NF-κB), were measured by enzyme-linked immunosorbent assays and immunoblot analyses.

Results

Eupatilin and jaceosidin blocked carrageenan-induced increase in leukocyte number and protein levels in air pouch exudates. Eupatilin and jaceosidin inhibited COX-2 expression and NF-κB activation, and markedly reduced TNF-α, IL-1β, and prostaglandin E2 (PGE₂) levels. They also inhibited hind paw edema induced by carrageenan. Eupatilin and jaceosidin had similar activity.

Conclusions

These findings suggest that eupatilin and jaceosidin may reduce inflammation by inhibiting NF-κB activation, and that Artemisia princeps inhibits inflammation because of these constituents.     

Antioxidant, anti-inflammatory, anti-nociceptive activities and composition of Lythrum salicaria L. extracts

Lythrum salicaria (purple loosestrife) known as "Tibbi hevhulma" in Turkish is used for its several beneficial health effects against as diarrhea, chronic intestinal catarrh, hemorrhoid and eczema in the form of a decoction or a fluid extract and to treat varicose veins, bleeding of the gums, hemorrhoid and eczema, externally. Dried herbal parts of Lythrum salicaria L. (Lythraceae) were sequentially extracted with different solvents such as petroleum ether, ethyl acetate, methanol and 50% aqueous methanol. Water extract of Lythrum salicaria was also prepared under reflux. Antioxidant, anti-inflammatory and anti-nociceptive activities of all the extracts were investigated using in vitro and in vivo methods, respectively. Free radical scavenging activity (1,1-diphenyl-2-picrylhydrazyl, DPPH* assay), iron(III) reductive activity, capacity of the inhibition of linoleic acid peroxidation and MDA formation, anti-nociceptive activity (p-benzoquinone-induced abdominal constriction test) and anti-inflammatory activity (carrageenan-induced hind paw edema model) were used for all the extracts. In addition, the content of total phenolics, flavonoids and flavonols in all the extracts were determined with spectrophotometric methods. Results were compared with reference antioxidants via ascorbic acid, butylated hydroxytoluene, and gallic acid. Qualitative and quantitative compositions of all the extracts were analysed using a HPLC-PDA system. Polar fractions were found to be rich in flavonoids such as isovitexin and isoorientin.