In vivo antigiardial activity of three flavonoids isolated of some medicinal plants used in Mexican traditional medicine for the treatment of diarrhea

Mexican traditional medicine uses a great variety of plants in the treatment of gastrointestinal disorders such as diarrhea. In order to understand the properties of some of their chemical constituents, three flavonoids (kaempferol, tiliroside and (-)-epicatechin) isolated from Geranium mexicanum, Cuphea pinetorum, Helianthemum glomeratum, and Rubus coriifolius, were assayed to demonstrate their in vivo antiprotozoal activity; using an experimental infection of Giardia lamblia in suckling female CD-1 mice. Compounds tested showed antigiardial activity with values of ED(50) (micromol/kg) 0.072 for (-)-epicatechin, 2.057 for kaempferol and 1.429 for tiliroside. The most active flavonoid was the (-)-epicatechin, its activity was higher than metronidazole and emetine, drugs used as positive controls. In the case of kaempferol and tiliroside their potency was close to that of the metronidazole, but far less than emetine.     

Additional antiprotozoal flavonol glycosides of the aerial parts of Helianthemum glomeratum

Bioassay-guided fractionation of the methanol extract of aerial parts from Helianthemum glomeratum afforded five antiprotozoal flavonol glycosides: tiliroside, kaempferol-3-O-(3',6'di-O-E-p-coumaroyl)-betad-glucopyranoside, astragalin, quercitrin and isoquercitrin. The in vitro antiprotozoal assay showed that tiliroside was the most potent antiamoebic and antigiardial compound with IC(50) values of 17.5 microg/mL for Entamoeba histolytica and 17.4 microg/mL for G. lamblia. Isoquercitrin showed selectivity against E. histolytica (IC(50) 14.7 microg/mL) and quercitrin toward G. lamblia (IC(50) 24.3 microg/mL). All isolated compounds were less active than metronidazole and emetine, two antiprotozoal drugs used as positive controls.     

Antiprotozoal activity of the constituents of Rubus coriifolius

Extraction of the aerial parts of Rubus coriifolius, a medicinal plant used by the Maya communities in Southern Mexico to treat bloody diarrhoea, resulted in the isolation of seven known compounds (-)-epicatechin, (+)-catechin, hyperin, nigaichigoside F1, beta-sitosterol 3-O-beta-d-glucopyranoside, gallic acid and ellagic acid. All compounds were tested for their antiprotozoal activity against Entamoeba histolytica and Giardia lambia. Epicatechin was the main responsible for the antiprotozoal activity of the extract against both protozoa, its activity was comparable to emetine, but no exceeded that of metronidazole.     

Antiprotozoal properties of Helianthemum glomeratum

Structure characterization and biological evaluation of the compounds isolated from Helianthemum glomeratum, particularly that of the polyphenols, has been the aim of a series of studies carried out to define the further potential use of this plant in the treatment of infectious diarrhoea in children. The flavan-3-ols, (-)-epigallocatechin and (-)-epigallocatechin gallate, isolated from Helianthemum glomeratum roots were tested for their antiamoebic and antigiardial effects in vitro. Compared with the activity determined with the leaf and the root methanol extracts, the effect of (-)-epigallocatechin against Entamoeba histolytica was of a similar potency, nevertheless, it also suppressed the growth of Giardia lamblia in axenic cultures, a parasite that proved to be resistant to the crude extracts. It might be assumed that determined biological properties are due to the presence of (-)-epigallocatechin in the plant, although the flavonoids, kaempferol and tiliroside isolated from the leaves, could account for the antiprotozoal properties of this herbal resource, used in Mayan traditional medicine for the treatment of bloody diarrhoea.     

Anticonvulsant properties of the methanolic extract of Cyperus articulatus (Cyperaceae)

The methanolic extract of rhizomes of Cyperus articulatus, a plant used in traditional medicine in Africa and Latin America for many diseases, possesses anticonvulsant activity in mice. This extract protected mice against maximal electroshock (MES)- and pentylenetetrazol (PTZ)-induced seizures. It also delayed the onset of seizures induced by isonicotinic acid hydrazide and strongly antagonized N-methyl-D-aspartate-induced turning behavior. The ED(50) for protection against seizures was 306 (154-541) mg/kg intraperitoneally (i.p.) for the PTZ test and 1005 (797-1200) mg/kg i.p. for the MES test. The ED(50) of methanolic extract against N-methyl-D-aspartate-induced turning behavior was 875 (623-1123) mg/kg i.p. C. articulatus L. methanolic extract protected 54% of mice from seizures induced by strychnine at the dose of 1000 mg/kg i.p. but had no or a moderate effect only against picrotoxin- or bicuculline-induced seizures. With these effects, the rhizome of C. articulatus L. possesses anticonvulsant properties in animals that might explain its use as a traditional medicine for epilepsy in Africa.     

Screening for polyphenols,antioxidant and antimicrobial activitiesof extracts from eleven Helianthemum taxa (Cistaceae) used in folk medicine in south-eastern Spain

Ethnopharmacological relevance

The Helianthemum genus contains approximately one hundred taxa. Some of them are important medicinal plants used in several countries for many different purposes. However, studies addressing the phytochemistry of many of these species or their biological activities are currently nonexistent.

Aim of the study

To perform a comparative analysis of the qualitative composition of secondary metabolites and biological activities of the leaves of the most commonly distributed and used Spanish Helianthemum taxa in order to find a relationship between chemotype, biological activity and uses.

Materials and methods

water and 80% methanolic extracts derived from 11 different Helianthemum taxa were analyzed for their phytochemical composition using standard methods. Furthermore, HPLC analysis was performed for the estimation of gallic acid, ellagic acid, tannins and flavonols as marker compounds. The antioxidant activity was measured by employing the scavenging activity on DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonate) radicals. The 80% methanolic extracts were evaluated for antibacterial (Gram-positive Staphylococcus aureus, Enterococcus faecalis and Listeria monocytogenes and Gram-negative Escherichia coli and Salmonella enterica) activity using the micro dilution technique.

Results

The total polyphenolic content and antioxidant capabilities differed significantly among the Helianthemum leaf extracts. A strong correlation between total phenolic content, antioxidant properties and antimicrobial activity was found. The polyphenolic profile was specific for each taxon. Whereas Helianthemum alypoides, Helianthemum cinereum subsp. rotundifolium, Helianthemum hirtum, Helianthemum asperum, and Helianthemum marifolium subsp. marifolium were characterized by the presence of gallic acid, egallic derivatives and ellagitannins; the polyphenolic profile of Helianthemum apenninum subsp. cavanillesianum, Helianthemum syriacum and Helianthemum polygonoides was mostly based on flavonoids. Helianthemum cinereum, Helianthemum alypoides and Helianthemum marifolium consistently presented the strongest radical scavenging activity (in water extracts EC₅₀ ranges from 29.88 to 38.1 against DPPH and from 8.11 to 20.8 against ABTS, and in 80% MeOH extracts from 25.3 to 31.8 against DPPH and from 6.15 to 8.6 against ABTS), total phenol content (>117 mg GAE/l) and antimicrobial activities.

Conclusion

The Helianthemum taxa used in folk medicine did not cluster in a unique section, being equally distributed in two out of the four sections analysed. There was not a clear relationship between the chemotype, based on the polyphenolic composition of the taxa, and their taxonomical classification. However, the composition of the methanolic and water extracts from the leaves of plants belonging to the Helianthemum genus was strongly related to their medicinal uses.     

Antisecretory activity of plants used to treat gastrointestinal disorders in Mexico

Aqueous and methanolic extracts from 26 medicinal plants used in Mexico to treat gastrointestinal disorders were screened to evaluate their antisecretory activity on cholera toxin-induced intestinal secretion in rat jejunal loops model. Extracts were tested at a dose of 300 mg/kg. From 56 samples tested, both extracts from Chiranthodendron pentadactylon, Hippocratea excelsa and Ocimum basilicum were the most potent with inhibition values ranging from 68.0 to 87.6%. On the other hand, the methanolic extract of Geranium mexicanum (aerial parts) and the aqueous extract of Bocconia frutescens showed the highest activity with inhibition values of 93.4 and 86.0%, respectively. The results obtained in this study give some scientific support to the use of the Mexican medicinal plants employed for the treatment of gastrointestinal disorders such as diarrhea.     

Xanthine oxidase inhibitory activity of some Indian medical plants

Xanthine oxidase inhibitory activity was assayed from six species belonging to different families traditionally used for the treatment of gout and related symptoms by indigenous people of India. The aqueous, methanol-water mixture and methanolic extract of these plants were used for the experiment. Of the 18 extracts assayed, 14 extracts demonstrated xanthine oxidase inhibitory activity at 100 microg/ml, among which 10 extracts showed an inhibition greater than 50% and IC(50) values below 100 microg/ml. The methanolic extracts of Coccinia grandis, Datura metel, Strychnos nux-vomica and Vitex negundo showed more than 50% inhibition, hence, they were screened for their in vivo hypouricaemic activity against potassium oxonate-induced hyperuricaemia in mice. Methanolic extracts of Coccinia grandis and Vitex negundo showed a significant decrease in the serum urate level (3.90+/-0.07 mg/dl, P<0.001) and (6.26+/-0.06 mg/dl, P<0.01), respectively, when compared to hyperuricaemic control (11.42+/-0.14 mg/dl). This effect is almost similar to the serum urate level of allopurinol (3.89+/-0.07 mg/dl).     

Evaluation of the anti-inflammatory properties of Sclerocarya birrea (A. Rich.) Hochst. (family: Anacardiaceae) stem-bark extracts in rats

This study was designed to evaluate the anti-inflammatory effect of Sclerocarya birrea (family: Anacardiaceae) stem-bark aqueous and methanolic extracts in rats. Young adult, male Wistar rats (Rattus norvegicus) weighing 250-300g were used. The anti-inflammatory effects of aqueous and methanolic stem-bark extracts of the plant (SB, 500mg/kg p.o.) were examined on rat paw oedema induced by subplantar injections of fresh egg albumin (0.5ml/kg). Acetylsalicylic acid (ASA, 100mg/kg p.o.) was used as the reference anti-inflammatory agent for comparison. Both the aqueous and methanolic stem-bark extracts of S. birrea (SB, 500mg/kg p.o.) progressively and time-dependently reduced rat paw oedema induced by subplantar injections of fresh egg albumin. However, the methanolic extract of the plant produced relatively greater and more pronounced anti-inflammatory effect than its aqueous extract counterpart in the experimental animal model used. The two extracts of S. birrea stem-bark were found to be markedly less potent than ASA as anti-inflammatory agent. Although both the aqueous and methanolic extracts of S. birrea stem-bark are less potent than ASA as anti-inflammatory agent, the results of this experimental animal study indicate that the extracts possess anti-inflammatory activity, and thus lend credence to the suggested folkloric use of the plant in the management and/or control of arthritis and other inflammatory conditions in certain communities of South Africa.     

In vitro susceptibility of Entamoeba histolytica and Giardia lamblia to some medicinal plants used by the people of Southern Mexico

The antiprotozoal properties of six medicinal plants, used by the people of Southern Mexico to treat diarrhoea, were evaluated in vitroagainst Entamoeba histolyticaand Giardia lambliatrophozoites, the most active being the methanol extracts of Rubus coriifolius, Cuphea pinetorumand Helianthemum glomeratum. Further activity guided fractionation of Cuphea pinetorumroots extract led to the isolation of kaempferol and quercetin as the active principles. © 1998 John Wiley & Sons, Ltd.     

In vitro antiprotozoal activities and cytotoxicity of some selected Cameroonian medicinal plants

Eight extracts from seven selected Cameroonian medicinal plants, traditionally used to treat malaria and other protozoal diseases, were tested in vitro for their antiprotozoal activities against Plasmodium falciparum K1 chloroquine-resistant strain, Leishmania donovani, Trypanosoma cruzi and Trypanosoma brucei rhodesiense, protozoa responsible for malaria, visceral leishmaniasis, Chagas disease and African trypanosomiasis, respectively. The most active extract against Plasmodium falciparum K1 strain and Trypanosoma brucei rhodesiense was the methanolic extract of Albizia zygia (Fabaceae) stem bark with IC(50) values of 1.0 microg/ml and 0.2 microg/ml, respectively. Five extracts showed IC(50) values below 5mug/ml against Leishmania donovani, with the methanolic seed extract of Harungana madagascarensis showing the highest activity, but only the methanolic extract of Albizia zygia showed activity against Trypanosoma cruzi. Cytotoxicity and selectivity indexes were estimated for the most active extracts. The best ratio of cytotoxicity to antiplasmodial activity (SI(a)=14) was established for the methanolic leaf extract of Symphonia globulifera (Clusiaceae), while the methanolic stem bark extract of Albizia zygia showed the best ratio of cytotoxicity to antitrypanosomal activity (SI(b)=22.5).     

In vitro cytotoxic, antileishmanial and antifungal activities of ethnopharmacologically selected Gabonese plants

Seventy-seven crude extracts from leaves and stem barks of 15 Gabonese plants used in traditional medicine were evaluated for their cytotoxic, antileishmanial and antifungal activities. Most of the extracts exhibited cytotoxic activities toward human monocytes, and most particularly the hydromethanolic 50% (v/v) fraction of Ganophyllum giganteum leaves (IC(50)=1.3 microg/ml) as well as the methanolic extracts of Polyalthia suaveolens, Dioscorea preussii, Augouardia letestui leaves and Cola lizae stem barks (IC(50)<5 microg/ml). The methanolic extract of Polyalthia suaveolens displayed a strong antiproliferative activity against the promastigote form of Leishmania infantum parasites and presented a good antifungal activity on all the tested strains (IC(50)<1mg/ml). This extract was divided into six fractions: fraction F6 demonstrated a cytotoxic activity stronger than those of the crude extract (IC(50)=0.6 microg/ml), fractions F4 and F5 were devoid of cytotoxicity (IC(50)>100 microg/ml) and displayed interesting antileishmanial activity against the intracellular amastigote form of the parasite (IC(50)=5.6 and 12.4 microg/ml), respectively. However, the antifungal activity observed for the crude extract could not be recovered in the corresponding fractions.     

Effect of Mexican medicinal plant used to treat trichomoniasis on Trichomonas vaginalis trophozoites

Crude methanolic extracts from 22 Mexican medicinal plants were screened for antitrichomonal activity against Trichomonas vaginalis, which is the etiological agent of trichomoniasis. Among the plants tested Carica papaya and Cocos nucifera showed the best antitrichomonal activity with IC(50) values of 5.6 and 5.8 microg/ml, respectively. The extracts of Bocconia frutescens, Geranium mexicanum, and Lygodium venustum showed moderate activity with IC(50) values ranging from 30.9 to 60.9 microg/ml. All the other plant extracts were inactive (IC(50)>100 microg/ml). All extracts tested were less active than metronidazole (IC(50) 0.037 microg/ml), an antiprotozoal drug used as positive control. The results of the antiprotozoal screening support the popular uses of five of the plants tested for the treatment of some urogenital tract disorders in Mexican traditional medicine. However, seeds of Carica papaya and aerial parts of Bocconia frutescens should be used in herbal medicine with care to avoid toxicity.     

Antiulcerogenic activity of Zizyphus lotus (L.) extracts

Oral administration of aqueous extracts of Zizyphus lotus root barks (50-200 mg/kg) leaves (50-200 mg/kg) and fruits (200-400 mg/kg) produced a significant (p<0.01) and dose dependent inhibition to the acute ulcer induced by HCl/ethanol solution. The methanolic (MeOH), ethyl acetate (EtOAc) and chloroformic (CHCl(3)) leaves extracts when administered orally at the dose of 200 mg/kg, exhibited a significant (p<0.01) inhibition of gastric lesions by 45%, 76% and 33%, respectively. Indeed, methanolic and ethyl acetate root barks extracts significantly reduced the gastric lesions by 47% and 41%, respectively. While the chloroformic root barks extract had no significant activity (19%). The effect of all extracts was compared with cimetidine (100 mg/kg, 62%) and omeprazole (30 mg/kg, 93%). Volume, pH and acidity of gastric juice were studied in pylorus-ligated rats. Root barks (200 mg/kg, p<0.01), leaves (200 mg/kg, p<0.01) and fruits (400 mg/kg, p<0.05) aqueous extracts showed significant reduction of gastric juice secretion in pylorus ligated rats, whereas the other extracts did not show any significance. Thus, Zizyphus lotus extracts act essentially as cytoprotective agents, which support the antiulcer effect of this plant in the traditional medicine.     

Evaluation of antidepressant activity of methanolic and hydroalcoholic extracts of Acorus calamus L. rhizome through tail suspension test and forced swimming test of mice

ObjectiveAcorus calamus (AC) L. (Araceae) is an annual semi-aquatic and aromatic plant found in Europe, North America and Asia. Its rhizomes are often used by Native Americans, Americans, and Chinese as well as by other cultures. Ethnobotanical studies and documents have shown their use in various disease treatments, such as insomnia, mental disorders, diabetes mellitus, epilepsy, inflammation, asthma, neuropathic pain, and diarrhea. In this study, the antidepressant activity of methanolic and hydroalcoholic extracts of the AC rhizome part in mice was investigated.MethodsThree doses of methanolic extract of AC rhizome (MEACR) (25, 50 and 100 mg/kg b.wt), three doses of hydroalcoholic extract of AC rhizome (HAACR) (100, 200 and 400 mg/kg b.wt), and standards (imipramine, 15 mg/kg b.wt and fluoxetine, 20 mg/kg b.wt) was daily oral administration to the mice for consecutive 14 days. The extract effect on the immobility time was monitored by a tail suspension test (TST) and a forced swimming test (FST). Monoamine oxidase (MAO) levels were also analyzed using standard methods.ResultsThe optimum antidepressant activity was viewed at 100 mg/kg b.wt of MEACR extract and 400 mg/kg b.wt of HAACR extract with 23.82% and 20.59% immobility period reduction, respectively. Besides, the extracts weakened the FST-induced elevation of MAO activity significantly and returned to near-normal levels of neurotransmitters in the brain. 100 mg/kg b.wt or above of MEACR extract significantly prevented the MAO-A and MAO-B activities in mice brain at a dose-dependent fashion. But, just 400 mg/kg b.wt of HAACR extract prevented the activity of MAO-A and MAO-B. Fluoxetine and imipramine showed a tendency to prevent the activity of MAO-A and MAO-B.ConclusionThis study suggests that AC rhizome extract mediated antidepressant activity by modulating the central neurochemical and hypothalamic-pituitary-adrenal (HPA) axis in response to FST and TST-induced stress. Therefore, AC rhizome extract can be used as a valuable plant supplement to treat depressive disorders.     

In vitro susceptibility of Entamoeba histolytica and Giardia lamblia to plants used in Mexican traditional medicine for the treatment of gastrointestinal disorders

In our search for new antiprotozoal chemotherapy, we collected a selection of 26 plants used in Mexican traditional medicine for the treatment of gastrointestinal disorders. Methanolic extracts of these species were screened for their antiprotozoal activity against Entamoeba histolytica and Giardia lamblia trophozoites using in vitro tests. Among the tested extracts, the derivates of following species showed selectivity and significant antiprotozoal activity: Chiranthodendron pentadactylon, Annona cherimola and Punica granatum were the most active on Entamoeba histolytica with IC50 < 30 microg/ml. Dorstenia contrajerva, Senna villosa and Ruta chalepensis were the most active toward Giardia lamblia with IC50 < 38 microg/ml. The potency of Chiranthodendron pentadactylon (IC50 2.5 microg/ml) on Entamoeba histolytica was close that of to emetine, but far less than metronidazole, drugs used as control. The results of the antiprotozoal screening support the popular uses of the studied species for the treatment of diarrhoea and dysentery in Mexican traditional medicine.     

Evaluation of ethnobotanically selected Benin medicinal plants for their in vitro antiplasmodial activity

Twenty extracts from nine Benin medicinal plants, traditionally used to treat malaria, were screened for in vitro antiplasmodial activity towards Plasmodium falciparum K1 chloroquine resistant and 3D7 chloroquine sensitive strains. All plants showed antiplasmodial activity below 10 μg/ml. Nine extracts exhibited IC₅₀ values below 5 μg/ml towards one or both of the two strains. The most active extract towards the sensitive 3D7 strain was the methanolic extract of Croton lobatus aerial part, with an IC₅₀ value of 0.38 μg/ml. The best inhibition of the growth of Plasmodium falciparum resistant K1 strain was observed with the methylene chloride extract of Hybanthus enneaspermus and with the methanolic extract of Croton lobatus roots (IC₅₀=2.57 and 2.80 μg/ml, respectively).     

Hepatoprotective activity of Apium graveolens and Hygrophila auriculata against paracetamol and thioacetamide intoxication in rats

Seeds of Apium graveolens L. (Apiaceae) and Hygrophila auriculata (K. Schum.) Heine (Syn. Astercantha auriculata Nees, Acanthaceae) are used in Indian systems of medicine for the treatment of liver ailments. The antihepatotoxic effect of methanolic extracts of the seeds of these two plants was studied on rat liver damage induced by a single dose of paracetamol (3 g/kg p.o.) or thioacetamide (100 mg/kg, s.c.) by monitoring several liver function tests, viz. serum transaminases (SGOT and SGPT), alkaline phosphatase, sorbitol dehydrogenase, glutamate dehydrogenase and bilirubin in serum. Furthermore, hepatic tissues were processed for assay of triglycerides and histopathological alterations simultaneously. A significant hepatoprotective activity of the methanolic extract of the seeds of both the plants was reported.     

Screening for acetylcholinesterase inhibitory activity in plants used in Thai traditional rejuvenating and neurotonic remedies

Acetylcholinesterase (AChE) inhibitor has been used as a drug for the symptomatic treatment of Alzheimer's disease. In order to search for new AChE inhibitors, 32 plants used in Thai traditional rejuvenating and neurotonic remedies were collected. The plant methanolic extracts were tested for AChE inhibitory activity using Ellman's colorimetric method in 96-welled microplates. The results showed that the methanolic extracts from roots of Stephania suberosa Forman. and Tabernaemontana divaricata (L.) R.Br. ex Roem. & Schult. at concentration of 0.1 mg/ml inhibited more than 90% of AChE activity. At the same concentration, four extracts, i.e. stems of Piper interruptum Opiz., seeds of Piper nigrum L., rootbarks of Butea superba Roxb. and roots of Cassia fistula L. extracts showed 50-65% inhibitory activity on AChE. The rest of the extracts showed the AChE inhibitory activity below 50%.