Analgesic effects and anti-inflammatory properties of the crude methanolic extract of Schwenckia americana Linn (Solanaceae)

Aim of the study

To evaluate analgesic effect and anti-inflammatory properties of Schwenckia americana (Solanaceae), a medicinal plant used for treating rheumatic pains and swelling in North-western Nigeria.

Materials and methods

Three doses (25 mg/kg, 50 mg/kg and 100 mg/kg) of the crude methanolic extract of Schwenkia americana were evaluated for analgesic and anti-inflammatory activities using acetic acid induced writhing test, formalin induced nociception, and formalin induced hind paw oedema in rats.

Results

All doses (25, 50, 100 mg/kg) of the extract tested were effective. The extract at the tested doses produced a percentage inhibition of the acetic acid induced abdominal constriction of (53.3, 58.0 and 86.7%), respectively. A percentage inhibition of the formalin induced nociception of 44.00, 56.04, and 56.04% (early phase) and 33.00, 36.63 and 59.71% (late phase) was also produced. The inhibition of oedema formation increased with increasing dosage from 25 to 100 mg/kg. The crude extract produced a statistically significant analgesic and anti-inflammatory activity comparable to the effect of standard drug (10 mg/kg Piroxicam).

Conclusion

This study demonstrated the potential analgesic and anti-inflammatory properties of crude methanolic extract of Schwenkia americana thus justifying its traditional usage.     

Evaluation of antinociceptive effect of Aegiceras corniculatum stems extracts and its possible mechanism of action in rodents

Ethnopharmacological relevance

Aegiceras corniculatum (Linn.) Blanco is used in various traditional medicinal system(s) for the treatment of rheumatism, painful arthritis and inflammation. Therefore, the pharmacological studies of its antinociceptive effect was undertaken to validate its traditional use.

Materials and methods

n-Hexane, ethyl acetate and methanol extract(s) derived from Aegiceras corniculatum (stems) were studied using various nociceptive model(s) induced chemically or thermally in mice including acetic acid-induced writhing, formalin-induced paw licking and hot plate test.

Results

In acetic acid-induced writhing test, plant extracts dose dependently decreased the writhing numbers. The methanolic extract (1-10 mg/kg, i.p. in mice) of the plant was more potent than acetaminophen and acetyl salicylic acid, with an IC₅₀ of 4.2 ± 0.99 mg/kg. Moreover, the time of nociceptive behaviors induced by intraplantar formalin injection was also suppressed during 1st and 2nd phases in the presence of ethyl acetate extract whereas, n-hexane and methanolic extracts inhibited the paw licking in mice during the 1st (IC₅₀ 12 ± 0.76 mg/kg) and 2nd phases (IC₅₀ 3.8 ± 0.55 mg/kg). Naloxone, β-funaltrexamine, and naltrindole antagonized the n-hexane extract-induced antinociception in the first phase of formalin test indicating its non-selective analgesic response via opioid receptor(s). However, ethyl acetate extract was devoid of any opioid action. Additionally, these extracts significantly inhibited the pain stimulation in hot plate test. Withdrawal syndrome of morphine dependence was also diminished in the presence of plant extracts via potentiation of GABAergic system.

Conclusion

These results suggested that Aegiceras corniculatum extract(s) possesses analgesic properties and acts on the central nervous system, thereby suppressing the inflammatory pain justifying its folklore use.     

Antinociceptive activity of ethanolic extract and isolated compounds of Urtica circularis

Ethnopharmacological relevance

Urtica circularis (Hicken) Sorarú is a medicinal plant commonly used in traditional medicine to relieve pain in inflammatory processes.

Aim of the study

In the present study, the in vivo antinociceptive effect of Urtica circularis ethanolic extract and its isolated compounds has been investigated.

Materials and methods

Antinociceptive activity was evaluated through writhing, formalin and hot plate tests in mice. The phytochemical analysis was performed.

Results

The extract produced significant inhibition on nociception induced by acetic acid (ED50: 72.2 mg/kg, i.p.) and formalin (ED50: 15.8 mg/kg, i.p.) administered intraperitoneally and also orally. Atropine diminished the activity of the extract in the acetic acid test. In this model, at dose of 10 mg/kg i.p., vitexin was the most active of the isolated compounds (inhibition of 91%), and chlorogenic acid, caffeic acid and vicenin-2 (6,8-di-C-glucosyl apigenin) produced an inhibition of 72%, 41% and 41%, respectively, whereas apigenin did not show any activity.

Conclusions

These results suggest that Urtica circularis extract produced antinociception possibly related to the presence of vitexin, chlorogenic, caffeic acid and vicenin-2. The activation of cholinergic systems seems to be involved in the mechanism of antinociception of the extract.     

Effects of Scutellariae radix and Aloe vera gel extracts on immunoglobulin E and cytokine levels in atopic dermatitis NC/Nga mice

Aim of the study

The present study aimed to investigate the effects of Scutellariae radix (SR) and Aloe vera gel (AV), alone or in combination, on levels of immunoglobulin E (IgE) and inflammatory cytokines in spontaneous atopic dermatitis(AD)-like skin lesions.

Materials and methods

After spontaneous AD-like skin lesion was developed by adaptation to conventional conditions, mice were randomly assigned to control, SR (50 mg/kg, p.o.), AV (0.8 mg/kg, p.o.) and SRAV (50 mg of SR and 0.8 mg of AV/kg, p.o.) groups, and were treated for 6 weeks.

Results

SR and SRAV suppressed IL-5 levels compared with control, but had no effects on IgE levels (P < 0.05). AV increased IgE levels, but decreased both IL-5 and IL-10 compared with control (P < 0.05).

Conclusion

These results suggest that SR and AV modulate immunological responses in AD, mainly through influencing IL-5 or IL-10 levels.     

Evaluation of the use of Cocos nucifera as antimalarial remedy in Malaysian folk medicine

Ethnopharmacological relevance

White flesh extract of Cocos nucifera (coconut) was studied to ascertain the ethnopharmacological standing of its antimalarial usage in Malaysian folk medicine.

Materials and methods

The crude methanol extract was investigated for phytochemical constituents and acute oral toxicity. Antimalarial activity of different extract doses of 50, 100, 200 and 400 mg/kg were investigated in vivo against Plasmodium berghei (NK65) infections in mice during early, established and residual infections. Chloroquine (20 mg/kg) and pyrimethamine (1.2 mg/kg) were used as reference drugs.

Results

The results revealed that the extract contained some phytochemical constituents and is toxicologically safe by oral administration. The extract significantly reduced the parasitaemia by the 200 and 400 mg/kg doses in the all three in vivo assessment assays. However, the extract did not significantly increase the survival time of the infected mice.

Conclusions

The observed pharmacological activities suggest that the Malaysian folkloric medicinal application of Cocos nucifera has a pharmacological basis.     

The analgesic and anti-inflammatory effect of WIN-34B, a new herbal formula for osteoarthritis composed of Lonicera japonica Thunb and Anemarrhena asphodeloides BUNGE in vivo

Ethnopharmacological relevance

: Lonicera japonica Thunb and Anemarrhena asphodeloides BUNGE have been used for the treatment of a variety of inflammatory diseases, cold and infective diseases in many countries, including Korea and China.

Aim of the study

: This study aimed to assess the anti-nociceptive and anti-inflammatory activities of n-butanol fraction (WIN-34B) prepared from dried flowers of Lonicera japonica and dried roots of Anemarrhena asphodeloides as potential novel treatment of osteoarthritis.

Materials and methods

: Anti-nociceptive activities of WIN-34B (100, 200 and 400 mg/kg, p.o.) were measured using acetic acid-induced writhing response, formalin-induced paw licking, hot plate, radiant heat tail-flick, carrageenan-induced paw pressure, and Hargreaves tests, respectively. Anti-inflammatory activities of WIN-34B (100, 200 and 400 mg/kg, p.o.) were assessed using acetic acid-induced vascular permeability, carrageenan-induced paw edema, and croton oil-induced ear edema. Anti-osteoarthritis effect of WIN-34B was analyzed using monosodium iodoacetate (MIA)-induced osteoarthritis animal model.

Results

: WIN-34B exhibited better anti-inflammatory activity than that of celecoxib in carrageenan at the dose of 200 mg/kg and croton oil-induced paw edema and ear edema at the doses of 200 and 400 mg/kg. WIN-34B exhibited significant anti-inflammatory effects on vascular permeability. WIN-34B also exhibited significant anti-nociceptive activities in the late phase of formalin-induced paw licking and writhing response model in mice. In radiant heat tail-flick and carrageenan-induced paw pressure tests, WIN-34B at the dose of 400 mg/kg and at the doses of 200 and 400 mg/kg presented similar activities to indomethacin and celecoxib. Compared to indomethacin WIN-34B at 400 mg/kg showed similar or better anti-nociceptive activities after 1 and 2 h of theraphy in the hot plate test and better anti-nociceptive activity than that of celecoxib in Hargreves test. In the MIA-induced osteoarthritis animal models, WIN-34B at 400 mg/kg exhibited similar or better anti-nociceptive property than that of celecoxib throughout the pain measurement periods.

Conclusion

: When compared to celecoxib, WIN-34B exhibited similar or better anti-nociceptive and anti-inflammatory activities in osteoarthritic animal models, which may become a potential novel treatment for osteoarthritis.     

Anti-inflammatory effect of a methanolic extract of leaves of Dregea volubilis

Aim of study

The aim of this study was to establish the anti-inflammatory activity of the methanolic extract of Dregea volubilis leaves (MEDV) with its fractions and to delineate the possible mechanism of action for MEDV.

Materials and methods

The anti-inflammatory activities of MEDV along with its petroleum ether and chloroform fractions were evaluated in a carrageenan induced model of acute inflammation. The effect of MEDV on lipopolysaccharide induced production of nitric oxide (NO) in macrophages was also studied.

Results

MEDV (100, 200 and 400 mg/kg body weight) significantly reduced carrageenan induced paw edema; chloroform fraction was most potent (66%, p < 0.001). MEDV was non-toxic up to 125 μg/ml in mouse peritoneal macrophages wherein it (0-100 μg/ml) reduced lipopolysaccharide induced NO production.

Conclusion

MEDV possesses significant anti-inflammatory activity. Chloroform fraction of MEDV showed best anti-inflammatory activity.     

Analgesic, anti-inflammatory and antipyretic activities of the petroleum ether fraction from the ethanol extract of Desmodium podocarpum

Ethnopharmacological relevance

Desmodium podocarpum is a plant that has been used in the folk medicine to treat febrile diseases, cough and bleeding wounds. However, there is no scientific basis or reports in the modern literature regarding its effectiveness as an analgesic, anti-inflammatory and antipyretic agent.

Aims of the study

The objective of this study is to evaluate the analgesic, anti-inflammatory and antipyretic activities of the petroleum ether fraction (PEF) from the ethanol extract of Desmodium podocarpum.

Materials and methods

PEF (50, 100, 200 mg/kg) was estimated for its pharmacological properties by using the acetic acid-induced writhing test, the hot plate test, the Carrageenan-induced rat paw edema model, the dimethylbenzene-induced mouse inflammation model, and the lipopolysaccharide (LPS)-induced rat fever model. In addition, the acute toxicity of PEF was also studied.

Results

PEF significantly and dose-dependently inhibited the writhing responses in mice, increased reaction time of mice in the hot plate test, reduced carrageenan-induced paw edema in rats and the dimethylbenzene-induced ear edema in mice, and attenuated LPS-induced fever in rats. No death of mice was observed when orally administered PEF up to 4.2 g/kg.

Conclusions

These findings suggest that PEF possesses evident analgesic, anti-inflammatory and antipyretic activities, and has a favorable safety, which supports the use of Desmodium podocarpum as an analgesic, anti-inflammatory and antipyretic drug in the folk medicine.     

Antinociceptive and antiinflammatory activities of Adiantum latifolium Lam.: Evidence for a role of IL-1β inhibition

Aim of study

Adiantum, one of the most widely distributed genera of the family Pteridaceae, is employed in folk medicine worldwide. Adiantum latifolium Lam. has been used in Latin American traditional medicine as anxiolytic, analgesic and antiinflammatory. The present study investigates the antinociceptive and antiinflammatory properties of the methanolic extract of Adiantum latifolium (MEA) in animal models of pain and inflammation to confirm its medicinal use.

Material and methods

The antinociceptive and antiinflammatory activities of MEA were evaluated using the writhing, formalin, and tail-flick tests, carrageenan-induced paw edema and arachidonic acid-induced ear edema. Mice motor performance was evaluated in the rota rod test and the acute toxicity evaluated over 14 days.

Results

Intraperitoneal (1-100 mg/kg) or oral (100-400 mg/kg) administration of MEA produced a dose-related inhibition of acetic acid-induced writhing in mouse. Furthermore, treatment with MEA (100 mg/kg) inhibited both the early and late phases of formalin-induced hypernociception. In contrast, MEA (100 mg/kg/IP) did not prevent the thermal nociception in the tail-flick test. In addition, MEA (100 and 200 mg/kg/IP) inhibited important events related to the inflammatory response induced by carrageenan or arachidonic acid, namely local edema and increase in tissue interleukin-1β levels. MEA (300 mg/kg/IP)-treated mice did not show any motor performance alterations. Over the study period of 14 days, there were no deaths or toxic signs recorded in the group of mice given 1000 mg/kg of MEA.

Conclusion

The results demonstrate that Adiantum latifolium has antinociceptive and antiinflammatory activities, acting through the inhibition of IL-1β production.     

Antinociceptive and gastroprotective actions of ethanolic extract from Pluchea sagittalis (Lam.) Cabrera

Ethnopharmacological relevance

Pluchea sagittalis, an herbaceous plant widely distributed in South America, is used in folk medicine for the treatment of digestive diseases and inflammation.

Aim of the study

This study was designed to investigate the antinociceptive and gastroprotective effects of the ethanolic extract (EE) of aerial parts from Pluchea sagittalis in rodents.

Materials and methods

The antinociceptive effects of EE was evaluated in mice after oral administration in chemical tests (acetic-acid, glutamate and formalin) or by biting behavior following intrathecal administration of cytokines such as interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) in mice. Furthermore, rats were treated with EE and subsequently exposed to acute gastric lesions induced by 80% ethanol. Afterwards the gastric lesion extension and the mucus levels of gastric mucosa were measured.

Results

The oral administration of EE showed a dose-dependent inhibition of acetic acid-induced abdominal constrictions and glutamate-induced pain in mice, with ID₅₀ values of 624.0 (523.0-746.0) mg/kg and 368.0 (216.0-628.0) mg/kg, respectively. In the formalin test, the EE also produced significant inhibition of the inflammatory phase, with an ID₅₀ value of 411.0 (183.0-721.0) mg/kg; however, it was ineffective in the neurogenic phase caused by formalin. In addition, oral treatment with EE caused a significant inhibition of biting behavior induced by i.t. injection of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). The antinociception caused by the EE (300 mg/kg, p.o.) was not reversed by naloxone (1 mg/kg, i.p.) when assessed in the acetic acid writhing test. The EE (300-1000 mg/kg, p.o.) did not affect the motor coordination of animals in an open-field model. Oral treatment with the EE protected rats against gastric lesions induced by ethanol, with an ID₅₀ value of 55.0 (46.6-64.9) mg/kg, and increased the mucus levels of gastric mucosa to levels found in the non-lesioned group.

Conclusions

The mechanism by which the extract produced antinociception still remains unclear, but this effect seems to be primarily related to the modulation or inhibition of the action of pro-inflammatory mediators. Furthermore, these data support, at least in part, the ethnomedical use of Pluchea sagittalis.     

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.     

Antidiabetic effect of Symplocos cochinchinensis (Lour.) S. Moore. in type 2 diabetic rats

Aim

Symplocos cochinchinensis (Lour.) S. Moore. is used in Indian system of traditional medicine to treat diabetes mellitus. The present study aims to investigate the antidiabetic efficacy of the hexane extract of Symplocos cochinchinensis leaves in high fat diet-low streptozotocin (STZ) induced type 2 diabetic rats.

Materials and methods

The doses for the study were fixed based on Irwin test. The hypoglycemic effect of the hexane extract of Symplocos cochinchinensis leaves were studied in normal rats. Oral glucose and insulin tolerance tests were carried out. The antihyperglycemic effect of the hexane extract at 250 and 500 mg/kg was studied in high fat diet-low STZ induced type 2 diabetic rats for 28 days.

Results

The extracts showed no adverse effects up to 5 g/kg concentration. In hypoglycemic study, after treatment with hexane extract at 250 and 500 mg/kg the blood glucose was mildly reduced. In oral glucose tolerance test, the treatment with the hexane extract at 250 and 500 mg/kg showed a highly significant reduction of 12.07% and 23.58% in plasma glucose levels, respectively 30 min after glucose load. The insulin tolerance test also showed improved insulin sensitivity after 60 min of insulin treatment. In high fat diet-low STZ induced type 2 diabetic rats, after 28 days treatment with the hexane extract at 250 and 500 mg/kg reduced the plasma glucose level by 17.04% and 42.10%, respectively. A significant reduction in plasma insulin, plasma and hepatic total cholesterol (TC), triglycerides (TG) and free fatty acids (FFA) and a significant increase in liver glycogen were observed in treated diabetic rats.

Conclusion

This study demonstrated the potential antidiabetic property of hexane extract of Symplocos cochinchinensis leaves on type 2 diabetes mellitus, thus justifying its traditional usage.     

Evaluation of anti-migraine potential of Areca catechu to prevent nitroglycerin-induced delayed inflammation in rat meninges: possible involvement of NOS inhibition

Ethnopharmacological relevance

Areca catechu nut extract is a popular folk remedy for the treatment of migraine in Kerala and Tamil Nadu states of India.

Aim of the study

In order to prove the claimed utilization of plant, the effect of hydroalcoholic extract of Areca catechu nut (ANE) was investigated in nitroglycerine induced inflammation in rat meninges. In these models infusion of nitric oxide donor glyceryl trinitrate (GTN) produces augmented plasma protein extravasation (PPE) in dura mater, provides an important substrate for the development of migraine in rats.

Materials and methods

The effect on plasma protein extravasation was assessed in both the models of intravenous and topical GTN application following oral administration of ANE (250 mg/kg and 500 mg/kg) in both curative and preventive treatment and compared with that of control positive. The l-NAME (15 mg/kg, i.v.) was used as reference standard. Plasma protein extravasation was measured using fluorescein as marker and was measured using a Perkin-Elmer LS-30 luminescence spectrometer.

Results

Expression of iNOS in the spleen after intravenous injection produced PPE into the dura mater in control positive group was significantly (P < 0.01) reduced to 1.553 ± 0.02499 and 1.398 ± 0.01887 by preventive treatment with ANE at the dose of 250 and 500 mg/kg, orally, respectively. The extravasation produced by topical GTN due to expression of iNOS in dural macrophages was also reduced to 1.555 ± 0.03384 and 1.425 ± 0.01204 by preventive treatment with ANE at the dose of 250 and 500 mg/kg, orally, respectively. While ANE do not showed any significant results in curative treatment in both the models of i.v. and topical GTN application.

Conclusion

These findings collectively indicate that the extract exhibited significant inhibition of iNOS, which may be the probable mechanism for its anti-migraine activity, providing evidence, at least in part, for its folkloric use.     

Polysaccharide fractions of Caesalpinia ferrea pods: potential anti-inflammatory usage

Ethnopharmacological relevance

Caesalpinia ferrea (Caesalpinioideae), known as pau-ferro or juca, has been used in the traditional medicine in North and Northeast of Brazil in inflammatory disorder, among others. Thus, experimental evaluation of the anti-inflammatory activity of extracts and fraction polysaccharides of Caesalpinia ferrea pods, and correlation with its anti-inflammatory activity and popular use is important.

Materials and methods

Total polysaccharides (TPL) were applied to ion exchange chromatography and eluted stepwise. Paw edema was induced s.c. by λ-carrageenan, dextran, histamine, serotonin, compound 48/80, bradykinin, prostaglandin E₂ (PGE₂) or l-arginine and analyzed by plethysmometry and protein leakage by spectrophotometry. Peritonitis was induced i.p. by carrageenan or N-formyl-methionyl-leucyl-phenylalanine (fMLP) and analyzed 4 h later for leukocyte migration and protein leakage. Animals were treated i.v. with TPL or polysaccharide fractions (0.01, 0.1, 1 mg/kg) 30 min before stimuli in both models. Toxicity (variation of body/organ mass and hematological/biochemical parameters) was evaluated after the seven-day treatment with the most active polysaccharide fraction (1 mg/kg; i.v.).

Results

Chromatography of TPL (2.8% yield) provided three major polysaccharide fractions (FI, FII, FIII). At 1 mg/kg, TPL inhibited the paw edema induced by carrageenan (60%) and FIII (fraction presenting high carbohydrate and low protein content) inhibited the inflammatory parameters in the paw edema induced by the following stimuli: carrageenan (70%), dextran (53%), histamine (65%), serotonin (62%), bradykinin (60%), PGE₂ (63%), nitric oxide (61%) and compound 48/80 (36%). Additionally, FIII at 1 mg/kg inhibited the carrageenan-induced edema in animals with intact mast cells, but only the late phase of those with degranulated mast cells elicited by compound 48/80. Moreover, FIII inhibited cell migration and protein leakage in the model of peritonitis elicited by carrageenan (88%) and fMLP (64%), being well tolerated by animals.

Conclusions

Extracts and polysaccharide fractions of Caesalpinia ferrea pods exhibit potent anti-inflammatory activity via negative modulation of histamine, serotonin, bradykinin, PGE₂ and NO released in the carrageenan-induced edema, showing involvement of mast cells. FIII could be interfering not only in the vascular, but also in cellular inflammatory events, revealing to be an important active component of traditionally prepared remedies used to treat inflammatory states.     

Increased sexual motivation in female rats treated with Humulus lupulus L. extract

Aim of the study

To evaluate the influence of Humulus lupulus extract on sexual behavior in female rats.

Materials and methods

Ovariectomized rats hormonally primed with estradiol benzoate (1.5 μg/rat) and progesterone (500 μg/rat) were acutely treated by oral gavage with Humulus lupulus extract dosed at 5, 10 and 25 mg/kg and then tested for partner preference and sexual receptivity.

Results

The administration of Humulus lupulus extract at the highest dose significantly increased the preference for the stimulus male during the partner preference test and the number of proceptive behaviors during the receptivity test, without affecting the lordosis response.

Conclusions

Humulus lupulus extract increased sexual motivation in hormone-primed female rats.     

Anti-inflammatory and analgesic activity of different extracts of Commiphora myrrha

Aim of the study

This present study was carried out to evaluate the anti-inflammatory and analgesic effects of 85% ethanol extract (EE) of Commiphora myrrha and its different fractions partitioned with petroleum ether extract (EPE), ethyl acetate extract (EEA), n-butanol extract (EBu), and the water extract (ECY). Moreover, the chemical constituents in EPE were analyzed and identified by UPLC-QTOF/MS/MS.

Materials and methods

The anti-inflammatory activities were investigated by utilizing the paw edema mice induced by formalin. In addition, we determined the levels of PGE₂ in the edema paw. While the analgesic activity was examined against thermally and chemically induced nociceptive pain in mice, using the acetic acid and hot-plate test methods. The effects of the administration of dolantin or indomethacin were also studied for references. The components in EPE were analyzed by the ultra-performance liquid chromatography coupled with mass spectrum.

Results

In the anti-inflammatory test, EE inhibited the development of paw swelling induced by formalin significantly. The pharmacological activities of the petroleum ether fraction (EPE) were stronger than the EE extract and other fractions at the dose of 100 mg/kg, and furthermore significantly decreased the levels of inflammatory factor PGE₂ in the edema paw tissue at the fourth hour after formalin injection. It has been also shown that the ethanol extract (EE) significantly reduced acetic acid-induced writhing response in mice at the dose of 200 mg/kg, and 100 mg/kg. The petroleum ether fraction (EPE) showed significant analgesic activity in the model at the dose of 100 mg/kg (p < 0.01), and the ethyl acetate fraction (EEA) exhibited less analgesic activity (p < 0.05). All test samples showed no significant analgesic activity on the hot plate pain threshold in mice. The UPLC-MS/MS chromatogram analysis of EPE stated that EPE contains the ingredients of sesquiterpenes, diterpenes, and diterpenic acids. Moreover, seven main compounds were identified.

Conclusion

These data demonstrated that the EE and EPE posses analgesic and anti-inflammatory activities and may support the fact the traditional application of this herb in treating various diseases associated with inflammatory pain.     

Antinociceptive activity of methanolic extract of Acmella uliginosa (Sw.) Cass

Ethnopharmacological relevance

Acmella uliginosa (Sw.) Cass. is a medicinal herbaceous plant that is commonly used by the Malay community in Malaysia to relieve pain often associated with mouth ulcers, toothache, sore throat, and stomach ache.

Aim

The study was carried out to investigate the antinociceptive effect of the methanolic extract of A. uliginosa (Sw.) Cass. flowers (MEAU) using murine models of chemicals and thermal nociception.

Materials and methods

Chemicals (acetic acid-induced abdominal constriction and formalin-, capsaicin-, glutamate-induced paw licking test) and thermal models (hot plate test) of nociception in mice were employed to evaluate the MEAU analgesic effect. The extract was given via oral administration at doses of 3, 10, 30 and 100 mg/kg.

Results

It was demonstrated that MEAU produced significant antinociceptive response in all the chemical- and thermal-induced nociception models, which indicates the presence of both centrally and peripherally mediated activities. Furthermore, the reversal of antinociception of MEAU by naloxone suggests the involvement of opioid system in its centrally mediated analgesic activity. Moreover, MEAU-treated mice did not show any significant motor performance alterations. No mortality and signs of toxicity were recorded following treatment of the MEAU.

Conclusion

The results from the present study appear to support the folkloric belief in the medicinal properties of A. uliginosa (Sw.) Cass. which against pain at both central and peripheral levels, in which the central antinociception is probably due to the participation of the opioid receptors.     

Evaluation of the sub-acute toxicity of the sclerotium of Lignosus rhinocerus (Cooke), the Tiger Milk mushroom

Ethnopharmacological relevance

Lignosus rhinocerus (known locally as ‘Tiger Milk mushroom’) is the most important medicinal mushroom used by the indigenous communities of Malaysia to treat fever, cough, asthma, cancer, food poisoning and as a general tonic. The sclerotium of the mushroom is the part with medicinal value. Lignosus rhinocerus was hitherto unexploited commercially because of limited supply. Recently, the mushroom was successfully cultivated.

Materials and methods

Sprague Dawley rats (5 rats/group/sex) were fed orally with 250, 500 and 1000 mg/kg TM02, 1000 mg/kg TM03 as well as 1000 mg/kg wild type Lignosus rhinocerus sclerotial powder. Sclerotial powder was orally administered once daily and consecutively for 28 days. Body weight of each animal was measured and any gross behavioral change was observed daily. Hematological and clinical biochemical parameters as well as histopathological analysis were carried out on 29th day.

Results

The results showed that oral administration of the sclerotial powder at daily dose of up to 1000 mg/kg had no adverse effect on the growth rate, hematological and clinical biochemical parameters (including renal and liver function parameters). Histological studies showed that the treatments did not induce any pathological changes in the liver, kidney, heart, spleen and lung of the animals.

Conclusion

In conclusion, our results show that there was no treatment-related sub-acute toxicity in rats following 28-days oral administration of 250, 500 and 1000 mg/kg TM02, 1000 mg/kg TM03 as well as 1000 mg/kg wild type Lignosus rhinocerus sclerotial powder. As the highest tested dose of 1000 mg/kg was not associated with any toxicity concern, the NOAEL dose is higher than 1000 mg/kg.     

Anti-inflammatory and anti-nociceptive effect of Betula platyphylla var. japonica in human interleukin-1β-stimulated fibroblast-like synoviocytes and in experimental animal models

Ethnopharmacological relevance

Traditional medicine has widely been used Betula platyphylla var. japonica to treat various inflammatory diseases including arthritis.

Aim of the study

To determine the anti-inflammatory, anti-nociceptive, and anti-arthritic effects of Betula platyphylla in interleukin-1β (IL-1β)-stimulated fibroblast-like synoviocytes from human rheumatoid arthritis and in nociceptive and inflammatory animal model.

Materials and methods

The inflammatory mediators such as IL-6, tumor necrosis factor (TNF)-α matrix metalloproteinase (MMP)-1, MMP-13, inducible nitric oxide synthesis (iNOS), nitrites, prostaglandin E₂ (PGE₂) and cyclo-oxygenase 2 (COX-2) activity of Betula platyphylla were tested in IL-1β-stimulated fibroblast-like synoviocytes. Tail withdrawal in response to thermal stimulation in tail flick test or paw flinching and shaking in response to sc hind paw formalin injection was measured 1 h after oral administration of Betula platyphylla. The former was evaluated with a paw pressure test, and the latter was measured using the squeaking score, and paw volume in inflammatory arthritis tests.

Results

Betula platyphylla significantly inhibited proliferation of IL-1β-induced synoviocytes. Betula platyphylla reduced the levels of inflammatory mediators, such as IL-6, TNF-α, MMP-1, MMP13, and PGE₂. In particular, Betula platyphylla significantly inhibited the releases of nitrites and iNOS, as well as release of NFκB, into the nucleus of IL-1β-treated synoviocytes, even at concentrations as low as 1 μg/ml. Oral administrant of Betula platyphylla at 400 mg/kg significantly decreased about 27.8% of tail flick withdrawal and inhibited about the number of paw flinches in both phases 1 and 2 of the formalin test. In the carrageenan-induced acute pain and arthritis model, Betula platyphylla dose dependently reduced the nociceptive threshold and the arthritic symptoms at day 8, respectively, and Betula platyphylla at 400 mg/kg markedly reduced the inflammatory area about 48% in the ankle joints. This capacity of Betula platyphylla at 400 mg/kg was similar to that of the celecoxib-2 inhibitor in carrageenan-induced nociceptive and inflammatory arthritis model.

Conclusions

These results suggest that Betula platyphylla has anti-nociceptive and anti-inflammatory effects in IL-1β-stimulated RA FLS and in an animal model of arthritis. Thus, the use of Betula platyphylla as a pharmaceutical candidate for the treatment of arthritis should be further studied.     

Orally administered mycelial culture of Phellinus linteus exhibits antitumor effects in hepatoma cell-bearing mice

Aim of the study

The aim of this study was to evaluate the anticancer effect of a mycelial culture from Phellinus linteus PL-7 (MCPL-7) and to elucidate its potential mechanism in vivo.

Materials and methods

SCID CB-17 mice received a transplant of Hep3B cells followed by daily MCPL-7 administrations for 8 weeks. Following tumor implantation, groups C-E were subcutaneously administered 50 mg/kg, 100 mg/kg, or 250 mg/kg MCPL-7 powder per day, respectively, for 8 weeks. Groups A and B received saline solution subcutaneously for 8 weeks.

Results

MCPL-7 administration induced a significant reduction in tumor size and was associated with a significant increase in T cell numbers; IL-12, IFN-γ and TNF-α secretion; NK cell activity; and phagocytic ability. Therefore, increased numbers of CD4⁺cells could have been caused by greater numbers of dendritic cells and macrophages in the spleen. Furthermore, the activation of dendritic cells and macrophages resulted in increased IL-12 secretion, which could upregulate NK cell activation. The increased secretion of IL-12, IFN-γ, and TNF-α enhanced the activity and phagocytic ability of NK cells. Thus, MCPL-7 may provide a potential therapeutic approach for both immunomodulatory and antitumor effects.