Diuretic activity of some Smilax canariensis fractions

Ethnopharmacological relevance

Smilax canariensis is an endemic species of the Canary Islands, popularly known as “Zarzaparrilla sin espinas”. The rhizome, leaves and stem of this species has wide use in folk medicine practice on the islands, where they are habitually employed as diuretic.

Aim of the study

In this paper we report on the diuretic activity in experimental animals of several fractions of the methanol extract of this species.

Materials and methods

Four fractions of the methanol extract of the rhizomes, leaves and stem of Smilax canariensis (50 and 100 mg/kg), Furosemide and Hydrochlorotiazide (10 mg/kg), were orally administered to laboratory animals to evaluate their diuretic activity. Water excretion rate, pH, conductivity, and content of Na⁺, K⁺ and Cl⁻ were measured in the urine of saline-loaded mice.

Results

All the studied extracts showed an interesting increase of the diuresis, although the n-buthanol (27%; p < 0.05) and ethyl acetate extract (35%; p < 0.01), at 100 mg/kg p.o., showed the most interesting diuretic activity, which suggested that this diuretic effect is associated with the compounds contained in the fractions of intermediate polarity (ethyl acetate and n-butanol), decreasing in the most extreme apolar and polar sub-extracts (dichloromethane and methanol: water respectively). The increase in diuresis produced by these two extracts was very close to the values of Hydrochlorothiazide (32%) or Furosemide (39%), used as reference diuretics.

Conclusions

These data, together with previous results on the aqueous and methanol extracts, reaffirm assertions made regarding the effectiveness of the extracts of this plant against urinary pathologies in the Canary Islands folk medicine.     

Immunomodulatory potential of Enicostema axillare (Lam.) A. Raynal, a traditional medicinal plant

Ethnopharmacological relevance

Enicostema axillare (Lam.) A. Raynal., (Gentianaceae) has been used in traditional Indian system of medicine as depurative and for the treatment of psoriasis, intermittent fever and cancer. Ethnobotanical survey conducted in Theni District indicated a high consensus for this plant as blood purifier and to treat dermatopathy and venereal infections. The present study evaluated the immunomodulatory activity of the methanol extract of Enicostema axillare.

Materials and methods

In vivo immunomodulatory activity of Enicostema axillare methanol extract (100 and 200 mg/kg b.w) was evaluated by assessing its effect on the total and differential leukocyte count, organ weight, hemaggultinating antibody titer, plaque forming cells, quantitative hemolysis of SRBC and delayed type hypersensitivity. Sheep red blood cells (SRBC, 5 × 10⁸ cells/0.1 mL) were used to immunize the animals. In vitro immunomodulatory potential of the extract was studied using peritoneal macrophages by evaluating its effects on NBT reduction, NO production and cytokine release.

Results

The animals treated with Enicostema axillare methanol extract showed a significant (P ≤ 0.05) increase in weight of the thymus and spleen. The total leukocyte and lymphocyte count was increased significantly (P < 0.005) by the treatment. There was no significant alteration in neutrophil count. A dose dependent increase in antibody titer value was observed. A decreased response to DTH reaction induced by SRBC was recorded. A potential phagocytic response was seen on treatment with the extracts at 10 and 25 μg/mL. The extract inhibited the release of pro-inflammatory cytokines and production of NO significantly in a dose dependent manner.

Conclusion

These findings suggested that the methanol extract of Enicostema axillare acted on both humoral and cell mediated immune functions and decreased the release of pro-inflammatory cytokines in the peritoneal macrophages.     

Hibiscus vitifolius (Linn.) root extracts shows potent protective action against anti-tubercular drug induced hepatotoxicity

Ethnopharmacological relevance

The roots of Hibiscus vitifolius Linn. (Malvaceae) is used for the treatment of jaundice in the folklore system of medicine in India. This study is an attempt to evaluate the hepatoprotective activity of the roots of Hibiscus vitifolius against anti-tubercular drug induced hepatotoxicity.

Materials and methods

Hepatotoxicity was induced in albino rats of either sex by oral administration of a combination of three anti-tubercular drugs. Petroleum ether, chloroform, methanol and aqueous extracts of roots of Hibiscus vitifolius (400 mg/kg/day) were evaluated for their possible hepatoprotective potential.

Results

All the extracts were found to be safe up to a dose of 2000 mg/kg. Among the four extracts studied, oral administration of methanol extract of Hibiscus vitifolius at 400 mg/kg showed significant difference in all the parameters when compared to control. There was a significant (P < 0.001) reduction in the levels of serum aspartate amino transaminase, alanine amino transferase, alkaline phosphatase, lactate dehydrogenase, total and direct bilirubin, where as an increase was found in the levels of total cholesterol, total protein and albumin. Liver homogenate studies showed a significant increase in the levels of total protein, phospholipids and glycogen, and a reduction in the levels of total lipids, triglycerides, and cholesterol against control animals. In the tissue anti-oxidant studies, we found a significant increase in the levels of catalase and superoxide dismutase, whereas there was marked reduction in the levels of thiobarbituric acid reactive substances, as compared to control. Histology of liver sections of the animals treated with the extracts showed significant reduction of necrosis and fatty formation when compared with control specimens.

Conclusion

These findings suggest that the root extracts of Hibiscus vitifolius have potent hepatoprotective activity, thereby justifying its ethnopharmacological claim.     

Assessment of the antidiarrhoeal properties of the aqueous extract, the 80% methanol extract and its soluble fractions of the leaves of Alstonia congensis Engl. (Apocynaceae) in Wistar rats

Aim of the study

To evaluate the antidiarrhoeal properties of Alstonia congensis leaves claimed to be effective for the treatment of diarrhoea by traditional healers during our ethnopharmacological investigation conducted in Kinshasa, Democratic Republic of Congo.

Materials and methods

The aqueous extract (decoction), and the 80% hot methanol extract (Soxhlet extraction) were obtained. This last extract was fractionated. The antidiarrhoeal activity was evaluated using castor oil and magnesium sulphate-induced diarrhoea in animals. The potential antibacterial activity of all samples was also assessed in vitro.

Results

At all oral doses of 100 and 200 mg/kg body weight, all A. congensis samples showed significant and dose-dependent antidiarrhoeal activity in treated Wistar rats characterised by significant increase of onset time and decrease of all other diarrhoeal parameters at various degrees compared to untreated groups in both models. At the highest oral dose of 200 mg/kg bodyweight, the 80% hot methanol and aqueous extracts produced 79.8±2.1% and 78.6±0.5%, and 75.0±2.1% and 71.4±2.1% inhibition of defecation and diarrhoea respectively against castor oil-induced diarrhoea, and 75.0±1.2% and 73.3±1.2% inhibition of diarrhoea respectively against magnesium sulphate-induced diarrhoea. The 80% hot methanol and aqueous detannified extracts showed low activity (42–47% inhibition of defecation and/or diarrhoea in both tests) suggesting that tannins may be responsible for the observed activity. At the same oral doses, the total alkaloid extract, the chloroform soluble fraction rich in alkaloids, the 80% methanol and the alkaline aqueous soluble subfractions produced more than 50% inhibition of defecation and/or diarrhoea in both tests. From the antibacterial testing in vitro, results indicated that all A. congensis samples exhibited an antibacterial activity mainly against bacteria implicated in diarrhoea with MIC and MBC values in the range of 15.6–500 μg/ml. The most active samples were the aqueous (decoction) and the 80% hot methanol dried extracts, the chloroform subfraction rich in alkaloids and the total alkaloid extract (MIC: 15.7–125 μg/ml, MBC: 31.2–250 μg/ml). Proteus varibilis was found to be the most resistant microorganism.

Conclusion

These reported results can partly support and justify the traditional use of extracts from Alstonia congensis leaves for the treatment of diarrhoea in tradittional medicine.     

Hyperpigmentant activity of leaves and flowers extracts of Pyrostegia venusta on murine B16F10 melanoma

Ethnopharmacological relevance

Pyrostegia venusta is a native Brazilian plant which has a variety of uses in traditional folk medicine including the treatment of vitiligo. However, its effectiveness on melanogenesis is not yet elucidated.

Aim of the study

This study aimed to investigate the melanogenic activity of hydroalcoholic extracts from the leaves and flowers of P. venusta on murine B16F10 melanoma cells.

Materials and methods

Different concentrations of the hydroalcoholic extracts of flowers and leaves of P. venusta were evaluated in trials of spontaneous melanin content (4 days), and cell viability by the MTT assay in murine B16F10 cells, and in the mushroom tyrosinase activity in vitro.

Results

Both extracts, leaves (0.1; 0.3; 1 and 3 μg/mL) and flowers (0.03 and 0.1 μg/mL) increased the melanin content in a concentration dependent manner after 4 days of incubation on melanoma cells. Leaves extract promoted enhancement of melanogenesis with maximum effect of 33.3 ± 3% (3 μg/mL), and the flower extract increased in 23.4 ± 3% (0.1 μg/mL). The cell viability test using MTT showed that in the same tested concentrations of both extracts no cell death was detected. Actually, either extract was not able to cause any change in the tyrosinase activity. HPLC analysis of P. venusta extracts found 0.09% and 1.08% of allantoin on leaves and flowers extracts, respectively.

Conclusions

The leaves and flowers extracts of P. venusta stimulates B16F10 melanogenesis at very low concentrations. These findings support the folk medicinal use of P. venusta on the treatment of hypopigmentation diseases, such as vitiligo.     

Investigation for anti-inflammatory and anti-thrombotic activities of methanol extract of Capparis ovata buds and fruits

Ethnopharmacological relevance

Capparis ovata Desf. has wide natural distribution in Turkey and it is consumed in pickled form. Flower buds, root bark, and fruits of the plant are used traditionally due to their analgesic, anti-inflammatory, wound healing, anti-rheumatismal, tonic, and diuretic effects.

Aim of the study

The aim of this study was to investigate the possible anti-inflammatory and anti-thrombotic effects of methanol extracts prepared from flower buds (CBE) and fruits (CFE) of C. ovata.

Materials and methods

Anti-inflammatory effects of CBE and CFE were assessed using carrageenan-induced and prostaglandin E₂-induced mouse paw edema models. For the anti-thrombotic effect evaluation, carrageenan-induced tail thrombosis model was performed in mice. The extracts were administered intraperitonally (i.p.) at the doses of 100, 200, and 300 mg/kg. The anti-inflammatory effect of Capparis extracts were tested in comparison to 10 mg/kg diclofenac and anti-thrombotic activity to 10 and 100 IU heparin.

Results

CBE at the doses of 200, and 300 mg/kg and CFE at the doses of 100, 200, and 300 mg/kg showed significant anti-inflammatory activity and CFE reached therapeutic concentration early than CBE in carrageenan inflammation model. In prostaglandin E₂ inflammation model, CBE and CFE exhibited significant inhibitory effects. The C. ovata extracts did not show remarkable anti-thrombotic effect.

Conclusions

Based on the results obtained, it can be concluded that fruits of C. ovata have more potent anti-inflammatory effect than flower buds. It has been suggested that inhibition of cyclooxygenase pathway is one of the mechanisms of the activity. C. ovata may be potentially used as therapeutic agents for inflammatory diseases.     

Antitussive,expectorant and bronchodilating effects of ethanol extract of Sorghum bicolor (L.) Moench roots

Ethnopharmacological relevance

Root of Sorghum bicolor (L.) Moench (RSB) is an herbal medicine in Traditional Chinese Medicine, still used in some rural areas in Central China as an alternative remedy to treat cough and asthma.

Aim of the study

The study was aimed at evaluating the antitussive, expectorant and bronchodilating effects of ethanol extract of RSB, support its folk use with scientific evidence, and lay a foundation for its further researches.

Materials and methods

RSB was extracted with 80% ethanol aqueous in reflux conditions, solutions were concentrated in reduced pressure, and lyophilized in vacuum to yield the RSB extract. Antitussive evaluations were carried out with three different models including ammonia liquor induced mice cough, capsaicin induced mice cough, and citric acid induced guinea pigs cough; phenol red secretion experiments in mice were performed to evaluate the expectorant ability; bronchodilating effects were evaluated with a bronchoconstrictive challenge induced by acetylcholine chloride and histamine in guinea pigs.

Results

In all the three antitussive tests, treatment of RSB significantly inhibited the frequency of cough, and prolonged the cough latent period in animals. And high dose of RSB (200 mg/kg in mice and 100 mg/kg in guinea pigs) created therapeutic activities as good as standard antitussive drug codeine phosphate (20 mg/kg). In the expectorant evaluation, 50, 100 and 200 mg/kg RSB treatment had significantly increased the amount of phenol red output for 0.39, 1.18, and 1.96 folds in mice tracheas. In the bronchodilating test, RSB treatment at 100 mg/kg extended the preconvulsive time for 44.84% compared with that of before treatment in guinea pigs.

Conclusions

RSB is an effective alternative medicine for the treatment of cough with potent antitussive, expectorant and bronchodilating activities.     

Antiulcerogenic effect of Croton urucurana Baillon bark

Ethnopharmacological relevance

Elephantopus mollis, Spilanthes africana, Urena lobata, Momordica multiflora, Asystasia gangetica and Brillantaisia ovariensis are used in Cameroonian traditional medicine for the treatment of bone diseases and fracture repair. The aim of this study was to evaluate the effect of ethanolic extracts of six Cameroonian medicinal plants on bone regeneration following bone and marrow injury.

Materials and methods

Ethanol extract of Cameroonian medicinal plants were administered (each extract at 250, 500 and 750 mg/kg doses) orally to adult female Sprague–Dawley rats having a drill hole injury (0.8 mm) in the femur diaphysis. Vehicle (gum-acacia in distilled water) was given to the control group. After 12 days of treatment, animals were euthanized and femur bones collected. Confocal microscopy of fractured bone was performed to evaluate bone regeneration (calcein labeling). Only active plant extracts were used for further experiments. Thus, callus was analyzed by microcomputed tomography. Osteogenic effects of the extracts were evaluated by assessing mineralized nodules formation of bone marrow stromal cells and osteoblast recruitment at drill hole site by immunohistochemistry.

Results

Ethanolic extract of the leaves and twigs of Elephantopus mollis (EM) and whole plant of Spilanthes africana (SA) dose-dependently stimulated bone regeneration at the drill hole site. EM at 250 and 750 mg/kg doses and SA at 750 mg/kg dose significantly increased mineral deposition compared to controls. Both extracts at 500 and 750 mg/kg doses improved microarchitecture of the regenerating bone evident from increased bone volume fraction, trabecular thickness, trabecular number, and decreased trabecular separation and structure model index. EM and SA extracts increased the formation of mineralized nodules from the bone marrow stromal cells. In addition, EM and SA extracts increased osteoblast recruitment at the drill hole site evident from increased Runx-2 positive cells following their treatments compared to control.

Conclusion

Ethanolic extracts of EM and SA accelerate fracture repair in rats via stimulatory effects on osteoblast differentiation and mineralization, thereby justifying their traditional use.     

Antinociceptive activity of aqueous extract and isolated compounds of Lithrea molleoides

Ethnopharmacological relevance

Lithrea molleoides (Vell.) Engl. (Anacardiaceae) is a medicinal plant commonly used in traditional medicine in South America.

Aim of the study

In the present study, the in vivo antinociceptive effect of L. molleoides' aqueous 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: 8.7 mg/kg, i.p.) and formalin (ED50: 7.7 mg/kg, i.p.) administered intraperitoneally and also orally. Yohimbine diminished the activity of the extract in the acetic acid test meanwhile haloperidol enhanced its effect. Two majority compounds, shikimic and vanillic acid were active in chemical nociceptive models used in this work, producing the highest inhibition of the writhing response at a dose of 30 mg/kg i.p. (55.4% and 57.1%, respectively) meanwhile at 100 mg/kg p.o. produced a slight response (23.3% and 23.9%, respectively).

Conclusions

These results suggest that L. molleoides' aqueous extract produced antinociception possibly related to the presence of shikimic and vanillic acid. The adrenergic and dopaminergic systems seem to be involved in the mechanism of antinociception of the extract.     

Anti-inflammatory and anti-nociceptive activities of Platanus orientalis Linn. and its ulcerogenic risk evaluation

Ethnopharmacological relevance

Leaves of Platanus orientalis Linn. are used in folk medicine as a wound-healer and ophthalmologic agent. Phytol derivatives from the leaves of plane-tree show anti-ulcer activity. Its analgesic and anti-inflammatory effects for knee pain were known to Persian scientists and hakims.

Materials and methods

The ethanolic extract of Platanus orientalis Linn. and its various fractions were given at a dose of 100 mg/kg po and 200 mg/kg po for testing their anti-inflammatory activity by carrageenan induced hind paw edema. The analgesic activity of the ethanolic extract and its fractions has been carried out by tail-flick method and writhing test at a dosage of 200 mg/kg po. Gastric ulceration studies have been further carried out to study the ulcerogenic risk evaluation of the ethanolic extract and its various fractions at a dose of 600 mg/kg body weight.

Results

Among the tested fractions, chloroform fraction exhibited better inhibition (68.33%) at 200 mg/kg po dosage when compared to the standard drug Ibuprofen (66.66%) after 3 h in the carrageenan induced hind paw edema. The ethanolic extract and all its fractions especially the chloroform (p<0.01) showed significant analgesic activity with insignificant ulceration as compared to the standard drug i.e. Ibuprofen. The histopathological study of ethanolic extract and its fractions revealed that none of them cause ulcer.

Conclusion

The present study indicates that Platanus orientalis Linn. has significant anti-inflammatory and analgesic effect.     

Antiulcer principle from Zingiber montanum

Ethnopharmacological relevance

Rhizome of Zingiber montanum has been extensively used as a folk medicine to ameliorate peptic ulcer at northern part of Bangladesh.

Aim of the study

To identify the antiulcer principle of the MeOH extract of the rhizome of Zingiber montanum by an ex vivo bioassay guided chromatographic separation and purification, and structure elucidation of the purified compound by spectroscopic methods.

Materials and methods

Dried powder of Zingiber montanum rhizomes was extracted with MeOH. The antiulcer activity of the crude extract and its chromatographic fractions were evaluated by the inhibition of 1 N HCl induced gastric lesions in Swiss albino mice. The pure compound was purified from the active fraction by crystallization with hexanes. Structure of the pure compound was elucidated by spectroscopic methods. The antiulcer activity of the pure compound was evaluated by the inhibition of 1 N HCl, 95% ethanol and indomethacin induced gastric lesions in mice.

Results

The MeOH extract of Zingiber montanum showed 61.97% and 83.10% inhibition of the 1 N HCl induced gastric lesions at doses of 200 mg/kg and 400 mg/kg, respectively, in mice. Chromatographic separation on silica gel of the extract was yielded seven fractions and the fraction 2 was found to have most potent antiulcer activity in mice. This fraction showed 77.46% inhibition of the 1 N HCl induced gastric lesions at a dose of 40 mg/kg in mice. Crystallization of the fraction yielded 1 (zerumbone, 180 mg). It showed statistically 45.77% and 92.25% inhibition of 1 N HCl induced gastric lesions in mice at doses of 20 mg/kg and 40 mg/kg, respectively. It also showed 29.07% and 45.35% inhibition of 95% ethanol induced gastric mucosal damage, and 64.76% and 72.38% inhibition of indomethacin induced gastric lesions in mice at doses of 20 mg/kg and 40 mg/kg, respectively.

Conclusion

Zerumbone (1) showed potent cytoprotective effect against necrotizing agent (HCl) and non-steroidal anti-inflammatory drug (indomethacin) induced gastric ulceration. It also exhibited moderate cytoprotective effect against noxious agent (EtOH) induced gastric lesions. It can be considered as a promising new antiulcer natural drug lead.     

In vitro anti-prostate cancer and ex vivo antiangiogenic activity of Vernonia guineensis Benth. (Asteraceae) tuber extracts

Ethnopharmacological relevance

Prostate cancer is a major problem worldwide and affects most men above the age of forty-five. Vernonia guineensis Benth. (Asteraceae) root decoction is used in folk medicine in Cameroon to treat a number of ailments including prostate cancer. The aim of this study was to provide a preliminary validation of the use of Vernonia guineensis Benth. extracts to treat prostate cancer by evaluating the in vitro activity of its crude extracts and isolated molecules on prostate cancer cells lines and effect on angiogenesis which is essential for growth and metastases of prostate cancer.

Materials and methods

Aqueous, dichloromethane and methanol extracts of Vernonia guineensis Benth. tubers were tested for activity against three prostate cancer cell lines (PC-3, DU-145 and AT3B-1). The dichloromethane extract was subjected to bioactivity guided fractionation. Anti-proliferation, clonogenic and antiangiogenic activity of the crude extracts and isolated compound were tested. The WST-1 assay was used for the anti-proliferation activity meanwhile the standard clonogenic test and the rat ring aorta assay were carried out to determine the clonogenic and antiangiogenic activity of tested products respectively.

Results

The aqueous and methanol extracts of Vernonia guineensis Benth. demonstrated weak activity against prostate cancer cell lines in vitro with IC₅₀ > 100 μg/mL. The dichloromethane extract was more potent with IC₅₀ of 56.233 ± 3.630 μg/ml and 67.316 ± 2.452 μg/ml against the DU-145 and PC-3 cell lines respectively. Activity guided fractionation of this extract yielded a Pentaisovalerylsucrose (1) isolated for the first time from a natural source to the best of our knowledge. Compound 1 demonstrated in vitro activity against the human prostate cancer cell lines PC-3 and DU-145 with IC₅₀ of 5.701 ± 0.142 μM and 4.275 ± 0.710 μM, respectively. The IC₅₀ of the compound was 5.763 ± 0.425 μM against AT3B-1, a rat prostate cancer cell line expressing P-glycoprotein which is linked to drug resistance in most metastatic cancers. Compared to compound 1, Paclitaxel and Docetaxel were active against AT3B-1 at 2.641 ± 1.253 μM and 0.613 ± 0.251 μM. Paclitaxel showed IC₅₀ values of 0.004 ± 0.002 μM and 0.003 ± 0.001 μM against DU-145 and PC-3 prostate cancer cell lines respectively. Docetaxel showed IC₅₀ values of 0.002 ± 0.001 μM and 0.004 ± 0.001 μM against DU-145 and PC-3 prostate cancer cell lines respectively.

Conclusion

The in vitro anti-prostate cancer and the antiangiogenic activity of Vernonia guineensis Benth. extracts and isolated compound support the use of the tubers of this plant for the treatment of prostate cancer.     

Anticonvulsant effect of Marsilea quadrifolia Linn. on pentylenetetrazole induced seizure: a behavioral and EEG study in rats

Ethnopharmacological relevance

Marsilea quadrifolia Linn (MQ) extract has been used traditionally as sedative and antiepileptic drug in India.

Aim of this study

To investigate the anticonvulsive potential of MQ extracts by using behavior and electroencephalographic (EEG) analysis on pentylenetetrazole (PTZ) induced seizure model in rats.

Materials and methods

For anticonvulsant effect, 60 minutes after administration of MQ, behavior and EEG were analyzed during PTZ (60 mg/kg) induced seizures. Changes of EEG power, latency of onset of seizure, seizure severity score, and duration of epileptic seizure were determined.

Results

Both the water and ethanol extract of MQ increased the latency of seizure but also decreased duration of epileptic seizure and seizure severity score. This reduction of seizure severity was also observed in EEG recording and EEG power analysis. The effectiveness of MQ ethanol extract is better than MQ water extract.

Conclusion

Both water and ethanol extract of MQ were effective in reducing the severity of behavioral and EEG seizures induced by PTZ in rats. This study justifies the traditional use of this plant in epilepsy.     

Anti-urolithiatic effects of Punica granatum in male rats

Ethnopharmacological relevance

The traditional use of Punica granatum has been reported to regulate urine discharge and controls the burning sensation of urine.

Materials and methods

Animals model of calcium oxalate urolithiasis was developed in male rats by adding ethylene glycol 0.75% in drinking water. The Punica granatum chloroform extract (PGCE) and Punica grantum methanol extract (PGME) orally at 100, 200 and 400 mg/kg, respectively, were administered along with ethylene glycol for 28 days. On 28 day, 24 h urine was collected from individual rats and used for estimation of urine calcium, phosphate and oxalate. The serum creatinine, urea and uric acid levels were estimated in each animal. The kidney homogenate was used for the estimation of renal oxalate contents. The paraffin kidney sections were prepared to observe the CaOx deposits.

Results

The ethylene glycol control (Gr.-II) had significant (P < 0.001 vs. normal) increase in levels of urine oxalate, calcium and phosphate, serum creatinine, urea and uric acid and renal tissues oxalates, as compared to normal (Gr.-I). The paraffin kidney sections show significant histopathological changes. The treatment of PGCE and PGME at 100, 200 and 400 mg/kg doses, significantly (P < 0.001 vs. control) decreased the urine oxalate, calcium and phosphate, renal tissue oxalates and serum creatinine, urea and uric acid, in EG induced urolithiasis after 28 days.

Conclusions

The PGCE and PGME at the doses of 400 mg/kg, found to be more effective in decreasing the urolithiasis and regeneration of renal tissues in male rats.     

Effects of Plantago major L. leaf extracts on oral epithelial cells in a scratch assay

Ethnopharmacological relevance

The present study was undertaken to evaluate the effects from different leaf extracts of the traditional medicinal herb Plantago major L. (plantain) on cell proliferation and migration in vitro, as a test for potential wound healing properties.

Materials and methods

Water and ethanol-based extracts were prepared from Plantago major fresh and dried leaves, and tested in vitro in a scratch assay with oral epithelial cells.

Results

The scratch assay produced reliable results after 18 h. Most of the tested extracts increased the proliferation/migration of the oral epithelial cells compared to the negative control. A concentration of 1.0 mg/mL (on dry weight basis) appears to be optimal regardless of type of extract, and among the alternatives, 0.1 mg/mL was always better than 10 mg/mL. Ethanol-based extracts with a concentration of 10 mg/mL had very detrimental effects on cell proliferation/migration. At the other two concentrations, ethanol-based extracts had the most beneficial effect, followed by water extracts of fresh leaves, ethanol plus water extracts of dried leaves and, finally, water extracts of dried leaves.

Conclusions

This study suggests that both the water extracts and the more polyphenol-rich ethanol-based extracts of Plantago major leaves have medicinal properties. Further research is, however, needed to determine what compounds are responsible for the wound healing effects.     

Hypoglycemic and antihyperglycemic effect of Begonia malabarica Lam. in normal and streptozotocin induced diabetic rats

Aim of the study

The stem of Begonia malabarica was used traditionally by the Malasar tribe to treat diabetes. To validate the hypoglycemic and antihyperglycemic effects of the hexane, ethylacetate and methanol extracts obtained from an ethnomedicinal plant, Begonia malabarica.

Materials and methods

The doses for the study were fixed based on Irwin test. The hypoglycemic effect of hexane, ethylacetate and methanol extracts of Begonia malabarica stems were studied in normal animals. The antihyperglycemic effect of the methanol extract was studied in streptozotocin induced diabetic rats.

Results

In normal rats the treatment with the methanol extract of Begonia malabarica had shown a highly significant reduction (16.54 and 34.47%) in plasma glucose levels from the 0 h values at the dose of 100 and 200 mg/kg respectively. In streptozotocin induced diabetic rats the body weight of the Begonia malabarica methanol extract treated animals had shown a significant increase (13.38% at 200 mg/kg) after 4 weeks treatment. The plasma glucose levels were reduced significantly by 46.57 and 50.20% after 4 weeks treatment at 100 and 200 mg/kg respectively. Likewise the absolute kidney weight was also reduced in a significant manner. After 25 days treatment the Begonia malabarica methanol extract treated animals had shown low fasting plasma glucose levels (54.29, 61.34% in 100 and 200 mg/kg) and reduced postprandial plasma glucose levels (54.23, 65.96% in 100 and 200 mg/kg) when compared with diabetic control values. Serum insulin levels and liver glycogen levels were increased to 40.04 and 42.18% in 200 mg/kg Begonia malabarica methanol extract treated animals respectively. The treatment with Begonia malabarica methanol extract did not change the triglycerides and total cholesterol levels. The urea and creatinine levels were also reduced significantly by this treatment. The reduction in SGPT levels indicated the absence of toxicity of Begonia malabarica extract at this dose level.

Conclusion

This study supports the use of Begonia malabarica by the Malasar tribe for the treatment of diabetes. Fractionation of this extract may yield novel prototypes to manage diabetes mellitus.     

Anti-nociceptive and anti-inflammatory effects of Croton crassifolius ethanol extract

Ethnopharmacological relevance

Croton crassifolius has been used to treat snake bites, stomach ache, sternalgia, joint pain, as well as pharyngitis, jaundice, and rheumatoid arthritis in traditional Chinese medicine. However, there is no scientific evidence which supports the use in the literature.

Aim of the study

To investigate the anti-nociceptive and anti-inflammatory effects of ethanol extract of C. crassifolius.

Materials and methods

Anti-nociceptive actions of C. crassifolius were assessed in mice using the hot-plate test, acetic acid-induced writhing test, and formalin test. Anti-inflammatory effects of C. crassifolius were determined in three animal models: acetic acid-induced capillary permeability accentuation in mice, carrageenan-induced edema of the hind paw in rats, and cotton pellet-induced granuloma formation in rats.

Results

Ethanol extract of C. crassifolius showed no significant anti-nociceptive activity in the hot-plate test. However, extract at dosages of 45, 90 and 180 mg/kg significantly reduced acetic acid-induced writhing by 28.89% (P<0.05), 38.37% (P<0.05), and 56.53% (P<0.001), respectively. The extract also caused marked dose-related inhibition of formalin-induced pain in the second phase (P<0.05 for 45 mg/kg, P<0.001 for 90 and 180 mg/kg extract). C. crassifolius extract at dosages of 45, 90 and 180 mg/kg significantly reduced acetic acid-induced capillary permeability accentuation in mice by 26.18% (P<0.05), 65.70% (P<0.001), and 79.19% (P<0.001), and suppressed carrageenan-induced paw edema by 21.28% (P<0.05), 30.69% (P<0.01), and 49.17% (P<0.001) at 6 h after carrageenan injection, respectively. 180 mg/kg of the extract also showed significant activity against carrageenan-induced paw edema at 4 h. At 90 and 180 mg/kg, the extract inhibited cotton pellet-induced granuloma formation in rats.

Conclusions

These results collectively demonstrate that the ethanol extract of C. crassifolius possesses peripheral anti-nociceptive and anti-inflammatory effects, providing evidence to rationalize the traditional use of C. crassifolius for the treatment of pain and inflammation.     

Sedative activity of hexane extract of Keampferia galanga L. and its active compounds

Ethnopharmacological relevance

It is well known that fragrance impacts behaviors and autonomic functions, and is increasingly used as relaxant, carminative, as well as sedative in aromatherapy. Kaempferia galanga L. is one of the popular traditional aromatic medicinal plants used in tropics and subtropics of Asia including China, Japan and Indochina.

Aim of the study

The aim of present study was to investigate sedative effects of hexane extract of Kaempferia galanga L.and 2 active aromatic compounds (compound 1: ethyl trans-p-methoxycinnamate and compound 2: ethyl cinnamate ) included in the extyract by means of inhalation in mice.

Materials and methods

Sedative activity was assessed by inhalation of the hexane extract and two major aromatic compounds.

Results

Inhalation of hexane extract at the doses of 1.5 and 10 mg showed significant reduction of locomotor activity, indicating considerable sedative and relaxant effects. Compound 1 and 2 were proved to possess sedative effects at 0.0014 mg and 0.0012 mg respectively.

Conclusions

These results suggest the promising application of Kaempferia galanga L. and its constituents in aromatherapy.     

Validation of use of a traditional remedy from Bridelia grandis (Pierre ex Hutch) stem bark against oral Streptococci

Aim of the study

Bridelia grandis (Pierre ex Hutch) (Euphorbiaceae) is a medicinal plant traditionally used in Cameroon by pygmies Baka as a remedy for oral cavity affection. Bioassay-guided stem bark extracts were investigated for their in vitro antimicrobial properties as well as their phytochemical constituents.

Materials and methods

The first extraction was carried out according to the traditional use. Further extractions were carried out with solvents of different polarity such as methanol (MeOH), ethyl acetate (EtOAc) and mixtures of MeOH–H₂O. The antimicrobial activity of the extracts against oral Streptococci was evaluated on the basis of the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) by the macrodilution method; the bacterial surface hydrophobicity was also evaluated.

Results

Water, methanol and mixtures methanol–water extracts, exhibited antibacterial activity with MIC between 0.5 and 2 mg/ml justifying the traditional use of Bridelia grandis stem bark for oral cavity affection. Preliminary phytochemical analysis was performed on the most active extract (methanol) using appropriate tests and well established analytical screening methods, such as TLC and RP-HPLC/DAD.

Conclusions

The data obtained indicate that tannins constitute the chemical family responsible for the biological activity.