Antimicrobial interactions of Khoi-San poly-herbal remedies with emphasis on the combination; Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus

Ethnopharmacological relevance

Plants are often combined in traditional herbal remedies to increase medicinal efficacy, thus this investigation provides some insight into the antimicrobial efficacies of selected combinations.

Aims of the study

The first aim was to scientifically validate antibacterial efficacy of plant mixtures that are traded within peri-urban centres of Cape Town (Western Cape, South Africa). This was followed by an in-depth evaluation of the most antimicrobially active mixture; Agathosma crenulata, Dodonaea viscosa with Eucalyptus globulus.

Materials and methods

Methanol and aqueous extracts of six plant mixtures were screened for antibacterial properties against two Gram-negative and two Gram-positive bacteria using the minimum inhibitory microdilution method. Thereafter, chloroform: methanol (1:1; v/v) extracts, essential oils and aqueous extracts of Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus were assayed for antimicrobial activity independently and in various combinations. The fractional inhibitory concentration indices (?FIC) were determined for double and triple plant combinations to establish antimicrobial interactions.

Results

From the six plant mixtures prepared by herbalists, a methanol extract derived from combining Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus showed the best antibacterial activity. The MIC values of 49 μg/ml for Staphylococcus aureus and Bacillus subtilis, and 98 μg/ml for Klebsiella pneumoniae and Escherichia coli were recorded. When Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus were mixed in various 1:1 combinations, mostly additive and synergistic interactions were noted. The most noteworthy synergistic (ΣFIC value 0.07) 1:1 combinations were observed for the chloroform: methanol extracts of Agathosma crenulata mixed with Eucalyptus globulus against Klebsiella pneumoniae, Staphylococcus aureus and Bacillus subtilis. When combined in a mixture of three plants (1:1:1), enhanced efficacy was evident against most of the pathogens, for both organic and aqueous extracts. The triple combination against Bacillus subtilis demonstrated the greatest synergy (ΣFIC values of 0.03).

Conclusions

The enhanced antimicrobial efficacy and synergistic interactions noted for some of the mixtures, particularly the combination of Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus support the Western Cape Khoi-San traditional medicinal practices of combining plants for enhanced efficacy.     

An antimicrobial evaluation of plants used for the treatment of respiratory infections in rural Maputaland,KwaZulu-Natal,South Africa

Ethnopharmacological relevance

Abundant availability of medicinal plants in the study area offers low cost health care, but scientific validation is needed in order to lend credibility to the traditional use against respiratory infections.

Aim of the study

This study focussed on determining the antimicrobial efficacies of 30 plant species (independently and in various combinations) used for respiratory related infections in rural Maputaland.

Materials and methods

In vitro minimum inhibitory concentration (MIC) assays were undertaken on dichloromethane-methanol (CH₂Cl₂: MeOH) and aqueous extracts, as well as the hydro-distilled essential oils (for aromatic plants). Selected plant parts were assessed for antimicrobial activity against a range of respiratory pathogens i.e. Cryptococcus neoformans (ATCC 14116), Klebsiella pneumoniae (ATCC 13883), Moraxella catarrhalis (ATCC 23246), Mycobacterium smegmatis (ATCC 14468) and Staphylococcus aureus (ATCC 6538). The sum of the fractional inhibitory concentrations (∑FIC) was determined for plants traditionally used in combination. Isobolograms represent MIC values for a selection of interactions where two plants were combined in various ratios.

Results

The most antimicrobially active aqueous extracts were that of Ozoroa obovata and Sclerocarya birrea (0.10 mg/ml) while organic extracts from Parinari capensis subsp. incohata and Tetradenia riparia demonstrated the most noteworthy (0.03 mg/ml) activity. Although both Lippia javanica and Eucalyptus grandis were by far the most popular plants traditionally used for respiratory infections, the antimicrobial activity was mostly only moderate. Furthermore, the traditional use in a 1:1 combination did not display strong antimicrobial interactions, but isobolograms demonstrate (against some test organisms) that when combined in various ratios, predominant additive interactions are evident where E. grandis was present in larger proportions. The combination of E. grandis with O. obovata demonstrated synergism against both C. neoformans and K. pneumoniae, with ∑FIC values of 0.34 and 0.28 respectively. Various ratios of these two plants also demonstrated a predominantly synergistic profile.

Conclusion

Although this in vitro study supports the traditional use of some plants independently and in combination for the treatment of respiratory ailments in rural Maputaland, results demonstrate that the traditional selection of plants in higher frequency do not necessarily correlate with higher antimicrobial efficacy.     

Plants traditionally used individually and in combination to treat sexually transmitted infections in northern Maputaland,South Africa: Antimicrobial activity and cytotoxicity

Ethnopharmacological relevance

Although medicinal plants are used extensively to treat sexually transmitted infections (STIs) in rural northern Maputaland, KwaZulu-Natal, the efficacy and safety of these plants have not previously been evaluated.

Aim of study

A study was designed to investigate the in vitro antimicrobial activity and cytotoxicity profiles of a selection (individual plants and selected combinations) of traditionally used plants in this study area.

Materials and methods

Aqueous and organic (dichloromethane: methanol, 1:1) extracts were prepared. Antimicrobial activity was assessed using the minimum inhibitory concentration (MIC) assay against the STI associated pathogens; Candida albicans ATCC 10231, Ureaplasma urealyticum clinical strain, Oligella ureolytica ATCC 43534, Trichomonas vaginalis clinical strain, Gardnerella vaginalis ATCC 14018 and Neisseria gonorrhoeae ATCC 19424. For the combination study, interactions were assessed using the fractional inhibitory concentration (ΣFIC). The plant species were assessed for safety using the 3-[4,5-dimethyl-2-thiazol-yl]-2,5-diphenyl-2H-tetrazolium bromide (MTT) cellular viability assay on the human embryonic kidney epithelial (Graham, HEK-293) cell line.

Results

For the antimicrobial studies, U. urealyticum was the most sensitive of the six test organisms, with the aqueous extract of Ranunculus multifidus (0.02 mg/ml) and the organic extract of Peltophorum africanum (0.04 mg/ml) being the most antimicrobially active plant species studied. Sclerocarya birrea was found to have the broadest spectrum of activity (mean MIC of 0.89 mg/ml). The only plant species to exhibit some degree of cytotoxicity against the kidney epithelial cell line was Kigelia africana (100 µg/ml), with 22% and 16% cell death for the aqueous and organic extracts, respectively. Of the 13 combinations studied, several synergistic combinations were evident, the most prominent being the combination of Albizia adianthifolia and Trichilia dregeana (aqueous extract) with an ΣFIC value of 0.15 against O. ureolytica. Synergistic interactions were observed regardless of the ratio of the aqueous mixtures of the two plants. Syzygium cordatum and S. birrea (aqueous extract) was also a combination of interest, demonstrating synergistic (ΣFIC=0.42) interactions against O. ureolytica. This combination, however, also displayed some cytotoxicity towards the human epithelial cell line.

Conclusion

This study demonstrated that anecdotal evidence of plant use does not always correlate with in vitro activity. Furthermore, the toxicological profiling is of utmost importance as if not combined in its correct ratio can lead to potential adverse effects.     

Dragon's Blood extract has antithrombotic properties, affecting platelet aggregation functions and anticoagulation activities

Ethnopharmacological relevance

Dragon's Blood from Dracaena cochinchinensis (Lour.) S.C. Chen (Yunnan, China), as a traditional Chinese medicinal herb, was shown to have certain antithrombotic effects. A new preparation process was used to extract effective components from Dragon's Blood. A 95% ethanol extract A (EA) and a precipitate B (PB) fraction were obtained and compared. Reliability of the preparation process was validated by pharmacodynamic experiments.

Materials and methods

A rat/mouse thrombosis and blood stasis model was developed for this study, and EA and PB effects on thrombosis, platelet functions and blood coagulation activities were analyzed.

Results

It was observed that the EA fraction had significantly better inhibitory effects than the PB fraction on thrombosis (p < 0.05), platelet aggregation function (p < 0.01) and anticoagulation activity (p < 0.05-0.01).

Conclusions

The results obtained here showed that EA fraction from Dragon's Blood contained pharmacologically effective compounds with antithrombotic effects, partially improving platelet function and anticoagulation activity.     

Pharmacological properties of Anagallis arvensis L. ("scarlet pimpernel") and Anagallis foemina Mill. ("blue pimpernel") traditionally used as wound healing remedies in Navarra (Spain)

Ethnopharmacological relevance

: Anagallis arvensis and Anagallis foemina are traditionally used in Navarra (Spain) for dermatological purposes regarding wound healing properties. In some cases they are also used to threat internal infections although they are known to be toxic at high doses.

Aim of study

: Due to lack of studies, we decided to evaluate the potential of the plants as wound healing remedies measuring antimicrobial and anti-inflammatory properties using in vitro procedures.

Materials and methods

Antimicrobial effects were studied against four bacteria and one fungus. Anti-inflammatory properties were measured in terms of COX-1 and -2 inhibition as well as superoxide radical scavenging capacity.

Results

Both species exerted antimicrobial and anti-inflammatory effects. The methanolic extract obtained from Anagallis arvensis seemed to produce the highest inhibition in Candida albicans (MIC = 0.31 mg/ml). Inhibition of COX-1 and -2 was also stronger for methanolic extracts whereas aqueous were revealed as better free radical scavengers.

Conclusions

The study reveals that both species posses antimicrobial and anti-inflammatory activities related to their ethnomedicinal uses.     

Exploring Tagetes erecta Linn flower for the elastase, hyaluronidase and MMP-1 inhibitory activity

Ethnopharmacological relevance

Traditionally Tagetes erecta Linn flower is claimed to treat skin diseases like sores, burns, wounds, ulcers, eczema and several other skin ailments. The aim of the present experiment was to evaluate the anti-wrinkle potential of standardized flower extract of Tagetes erecta.

Materials and methods

The Tagetes erecta extract and fractions were screened for hyaluronidase, elastase and matrix metalloproteinase (MMP-1) inhibitory activity compared with the activity of standard oleanolic acid. Syringic acid and β-amyrin were obtained from the extract and quantified through RP-HPLC. Also the compounds were evaluated for anti-wrinkle activity.

Results

The methanol extract showed significant (^aP < 0.05) hyaluronidase and elastase inhibition with IC50 of 11.70 ± 1.79 μg mL⁻¹ and 4.13 ± 0.93 μg mL⁻¹ respectively and better MMP-1 inhibition compared to standard oleanolic acid. The isolated compounds syringic acid and β-amyrin found to inhibit enzymes comparable to oleanolic acid. The RP-HPLC analysis revealed that good amounts of syringic acid and β-amyrin (2.30%, w/w and 0.06%) are present in Tagetes erecta.

Conclusion

Tagetes erecta flower showed effective inhibition of hyaluronidase, elastase and MMP-1. Therefore, this experiment further rationalizes the traditional uses of this plant, which may be useful as an anti-wrinkle agent.     

In vitro evidence of phyto-synergy for plant part combinations of Croton gratissimus (Euphorbiaceae) used in African traditional healing

Aim of the study

Despite the extensive traditional use of Croton gratissimus Burch. var. gratissimus for medicinal purposes, scientific studies validating the therapeutic properties of this indigenous plant are lacking. As the bark, roots and leaves of C. gratissimus are used separately as well as in combination, this study focused on determining antimicrobial efficacies of the plant parts independently and in combination to assess possible pharmacological interactions (e.g. synergy, antagonism).

Material and Methods

The hydro-distilled leaf essential oil and extracts of bark, root and leaf were comparatively assessed for antimicrobial activity by means of microdilution minimum inhibitory concentration (MIC). The fractional inhibitory concentrations (FIC) were determined for the leaf and root (1:1), bark and root (1:1), leaf and bark (1:1) combination. Isobolograms were plotted to demonstrate interactions between various ratios of the roots and leaves.

Results

The MIC and FIC results indicated variable efficacies for the various plant part combinations, the greatest of which was noted for Cryptococcus neoformans in the root and leaf combination (MIC 0.4 mg/ml and FIC of 0.4). Isobolograms indicated the greatest synergy for Bacillus cereus, Candida albicans and Cryptococcus neoformans.

Conclusion

The observed synergistic interactions clearly indicate that the reductionist approach may often be short-sighted and that biological activity may be improved through combination therapy, where different complex metabolic pools collectively contribute to the enhanced effect.     

Evaluation of hydro-alcoholic extract of Eclipta alba for its anticancer potential: an in vitro study

Ethnopharmacological relevance

Eclipta alba is traditionally used as hepatoprotective agent. The study was designed to explore its antiproliferative activity on liver and other related cancer.

Aim of the study

The present study was designed to assess and establish the role of Eclipta alba as anti-cancer agent using HepG2, C6 glioma and A498 cell lines as model system.

Materials and methods

Antiproliferative and cytotoxic effects of the Eclipta alba hydroalcoholic extract (EAE) was determined using MTT assay. The expression level of NF-kB was analysed by western blotting and RT PCR. Gelatin zymography was done for gelatinase matrix metalloproteinases (MMP-2 and 9) analysis.

Results

EAE inhibited the cell proliferation in dose dependent manner in HepG2, A498 and C6 glioma cell lines with an IC50 of 22 ± 2.9, 25 ± 3.6 and 50 ± 8.7 μg/ml, respectively. The expression of MMP (2 and 9) was down-regulated with EAE treatment. DNA damage was observed following 72 h of extract treatment, leading to apoptosis. Additionally, the expression level of NF-kB was evaluated with western blotting and RT-PCR and was found to be down-regulated/inactivated.

Conclusions

The data establish the existence of anti-proliferative, DNA damaging and anti-metastasis properties in EAE which is yet unexplored and hold high therapeutic impact.     

Wound healing and antimicrobial potential of Carissa spinarum Linn. in albino mice

Ethnopharmacological relevance

Carissa spinarum Linn. (Apocynaceae), popularly known as Jungle karunda, has been traditionally used as wound healing agent.

Aim of the study

To investigate in vivo wound healing and antimicrobial properties of methanolic extract of Carissa spinarum.

Materials and methods

The effect of methanolic extract of Carissa spinarum root extracted by cold maceration was evaluated on burn wound model in mice. The wound healing activity of 1% and 2.5% (w/w) extract was assessed by the rate of wound contraction, period of epithelization and hydroxyproline content. Histological study of the granulation tissue was carried out to know the extent of collagen formation in the wound tissue. The antimicrobial activity of extract was also studied against the bacterial and fungal strain using agar dilution method.

Results

The results showed that Carissa spinarum root extract has significant wound healing activity as evident from the rate of wound contraction and epithelization. Hydroxyproline expressions and histological parameters were also well correlated with the healing pattern observed. Methanolic extract also exhibited significant antimicrobial activity against all the tested microorganisms.

Conclusions

The present study provides a scientific rationale for the traditional use of Carissa spinarum in the management of wounds.     

The cardioprotective effects of marrubiin, a diterpenoid found in Leonotis leonurus extracts

Ethnopharmacological relevance

Leonotis leonurus L. (Lamiaceae) is used as a traditional medicine for a variety of ailments in South Africa. The diterpene marrubiin is the major product constituent in specimens of this plant occurring in South Africa.

Materials and methods

Marrubiin isolated from South African specimens of L. leonurus in addition to an organic extract of L. leonurus were tested in vivo, ex vivo and in vitro for their anticoagulant, antiplatelet and anti-inflammatory activities.

Results

Marrubiin and the organic extract suppressed coagulation, platelet aggregation and inflammatory markers. For the coagulation markers it was found that the organic extract and marrubiin significantly prolonged activated partial thromboplastin time (APTT). Fibrin and D-dimer formation were drastically decreased. These findings were observed in an ex vivo model and an obese rat model. Chemokines enhance leukocyte recruitment to inflammatory sites. TNF-α and RANTES secretion were significantly reduced by the extract and marrubiin when determined in the obese rat model relative to the controls. Calcium mobilization and TXB₂ synthesis were suppressed by the extract and marrubiin. An in vitro model was used to elucidate the antiplatelet mechanism and it was found that the extract and marrubiin inhibited platelet aggregation by inhibiting the binding of fibrinogen to glycoprotein (GP) IIb/IIIa receptor in a concentration dependent manner.

Conclusion

The findings reflect that marrubiin largely contributes to the extract's anticoagulant, antiplatelet and anti-inflammatory effects observed.     

Antimicrobial activity of southern African medicinal plants with dermatological relevance: From an ethnopharmacological screening approach,to combination studies and the isolation of a bioactive compound

Ethnopharmacological relevance

Ethnobotanical reports on more than 100 southern African medicinal plants with dermatological relevance have been highlighted, yet there is still limited scientific data to support claims for their antimicrobial effectiveness against skin pathogens. Guided by ethnobotanical data, this paper explores the antimicrobial efficacies of southern African medicinal plants used to treat skin ailments.

Aim of the study

To investigate the antimicrobial properties of southern African medicinal plants against dermatologically relevant pathogens. The study also aimed at providing a scientific rationale for the traditional use of plant combinations to treat skin diseases and the isolation of the bio-active compound from the most active species, Aristea ecklonii (Iridaceae).

Materials and methods

Organic and aqueous extracts (132) were prepared from 47 plant species and screened for antimicrobial properties against dermatologically relevant pathogens using the micro-titre plate dilution method. Four different plant combinations were investigated for interactive properties and the sum of the fractional inhibitory concentration (?FIC) calculated. Isobolograms were used to further investigate the antimicrobial interactive properties of Pentanisia prunelloides combined with Elephantorrhiza elephantina at varied ratios. A bioactivity-guided fractionation process was adopted to fractionate the organic leaf extract of Aristea ecklonii.

Results

Plants demonstrating notable broad-spectrum activities (MIC values ≤1.00 mg/ml) against the tested pathogens included extracts from Aristea ecklonii, Chenopodium ambrosioides, Diospyros mespiliformis, Elephantorrhiza elephantina, Eucalyptus camaldulensis, Gunnera perpensa, Harpephyllum caffrum, Hypericum perforatum, Melianthus comosus, Terminalia sericea and Warburgia salutaris. The organic extract of Elephantorrhiza elephantina, a plant reportedly used to treat acne vulgaris, demonstrated noteworthy antimicrobial activity (MIC value of 0.05 mg/ml) against Propionibacterium acnes. Similarly, Diospyros mespiliformis reported for its traditional use to treat ringworm, also displayed noteworthy antimicrobial activity against Trichophyton mentagrophytes (MIC 0.10 mg/ml) and Microsporum canis (MIC 0.50 mg/ml). The aqueous root extracts of Pentanisia prunelloides combined (1:1) with Elephantorrhiza elephantina displayed synergistic interactions (?FIC values 0.31–0.38) against Staphylococcus aureus, gentamycin–methicillin resistant Staphylococcus aureus, Staphylococcus epidermidis and Candida albicans. Fractionation of Aristea ecklonii resulted in the isolation of the known bio-active compound, plumbagin, displaying noteworthy antimicrobial activity (MIC range between 2.00 μg/ml and 16.00 μg/ml).

Conclusion

Most of the plant extracts demonstrated pathogen specific antimicrobial effects with a few exhibiting broad-spectrum activities. Positive antimicrobial effects noted for plants such as Elephantorrhiza elephantina and Diospyros mespiliformis used for acne vulgaris and ringworm infections, respectively, give some validation to their reported traditiona l uses. Synergistic interactions noted for Pentanisia prunelloides combined with Elephantorrhiza elephantina validate an enhanced antimicrobial effect when used in combination. Noteworthy antimicrobial activities (MIC range between 2.00 μg/ml and 16.00 μg/ml) were observed for plumbagin isolated from Aristea ecklonii.     

Acute and subchronic oral toxicity studies of Nelumbo nucifera stamens extract in rats

Ethnopharmacological relevance

Since the use of Nelumbo nucifera stamens in herbal medicines as well as in cosmetic products are highly prevalent in Thailand and increasing worldwide, acute and subchronic toxicity studies to confirm the safe use of Nelumbo nucifera stamens are warranted.

Aim of the study

Acute and subchronic oral toxicity studies of Nelumbo nucifera stamens extract in rats were performed in the present study in order to evaluate its safety.

Materials and methods

In acute toxicity study, Nelumbo nucifera stamens extract was administered by oral gavage to Sprague-Dawley rats (5 males and 5 females) at a dose of 5000 mg/kg. In subchronic toxicity study, the extract at doses of 50, 100, and 200 mg/kg/day were given orally to groups of rats (6 rats/dose/sex) for 90 consecutive days.

Results

The extract at a dose of 5000 mg/kg produced no treatment-related signs of toxicity or mortality in any of the animals tested during 14 days of the study. In the repeated dose 90-day oral toxicity study, there was no significant difference in body weight between the control and all treatment groups with the exception of the body weight of the female group treated with 200 mg/kg/day of the extract which was statistically significantly less than that of its control counterpart on day 90 but the percent weight changes of both groups were almost similar. Some statistically significant differences in hematological and biochemical parameters as well as in some internal organ weights of both male and female rats treated with the extract at the highest dose were observed. However, no abnormality of internal organs was observed in both gross and histopathological examinations.

Conclusions

These results suggest that the oral lethal dose of Nelumbo nucifera stamens extract for male and female rats is in excess of 5000 mg/kg and the no-observed-adverse-effect level (NOAEL) of the extract for both male and female rats is considered to be 200 mg/kg/day.     

Apoptotic activity of ethanol extract from Styrax Japonica Siebold et al Zuccarini in HepG2 cells

Ethnopharmacological relevance

Styrax japonica Siebold et al Zuccarini (SJSZ) has been used to heal inflammation and bronchitis as an herbal plant in Korea.

Aims of the study

The purpose of the present study is to determine whether the ethanol (EtOH) extract of SJSZ induces the programmed cell death (apoptosis) in human hepatoma HepG2 cells.

Materials and methods

It was evaluated cytotoxicity using MTT assay, amount of intracellular reactive oxygen species (iROS) and Ca²⁺ using fluorescence. Activities of apoptotic relevant factors [Bid, cytochrome c, caspase-9, -3, and poly-(ADP-ribose) polymerase (PARP)] were measured by Western blot.

Results

The results in this study indicated that ethanol extract of SJSZ (75 μg/ml) stimulates to increase amount of iROS, Ca²⁺, and the apoptotic relevant factors [Bid, cytochrome c, caspase-9, -3, and poly-(ADP-ribose) polymerase (PARP) in the HepG2 cells.

Conclusion

The results in this study indicated that ethanol extract of SJSZ (75 μg/ml) induces programmed cell death (apoptosis) in the HepG2 cells. Therefore, we speculate that ethanol extract of SJSZ could be used for healing of hepatocarcinoma as one of chemotherapeutic agents.     

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.     

Animals used for medicinal and magico-religious purposes in western Granada Province, Andalusia (Spain)

Ethnopharmacological relevance

This paper serves to document and make known the folk medicinal uses of animals and animal products in the western part of the province of Granada (Andalusia, Spain), analyzing not only the species used, the administration methods and the ailments treated, but also the cultural implications of this traditional knowledge and the large percentage of magico-religious or ritual practices.

Materials and methods

The information was gathered through semi-structured open interviews of a total of 42 people, as a part of an in-deep ethnobiological research in 16 different municipalities within the study area. The use-value index was calculated for highlighting the most important species for the population.

Results

In this part of Granada, 26 different animals have been employed in the treatment of 26 conditions from 10 different pathological groups, amounting 150 total reports (7% of the total ones including plant and animal medicinal resources). Although some of the uses were included in several classical and anthropological works, most remain unpublished. Cultural and environmental implications are discussed.

Conclusions

The traditional use of animals in medicine is low known for southern Europe. 26 species constitute a significant number of animals used, considering the socio-economic and cultural context in which the study area is situated, and its small size. Albeit the main part of the information presented in this paper can be more important in an historical or anthropological standpoint, most of the medicinal resources presented have not been deeply studied.     

Labisia pumila var. alata reduces bacterial load by inducing uroepithelial cell apoptosis

Ethnopharmacological relevance

Labisia pumila var. alata (LPva) is a traditional medicinal herb used by Malaysian women to treat many ailments of the genitourinary tract. Its phytoestrogenic properties suggest potential to prevent recurrent urinary tract infection (UTI) in women post menopause. The aim of this study was therefore to investigate the mechanisms of action of LPva in an in vitro model of UTI.

Materials and methods

Bladder epithelial cell lines T24 and 5637 and uropathogenic Escherichia coli (UPEC) strain CFT073 were used to model uroepithelial infection. The ability of LPva to induce programmed cell death was tested using the Annexin-V-FLUOS and TUNEL assays. Expression of caveolin-1, β1 integrin and antimicrobial peptides HBD-2 and LL-37 in response to LPva treatment and/or infection, was assessed using RT real-time PCR. Effects on protein expression were confirmed by Western blot analysis. Sensitivity and yeast agglutination assays were employed to determine if LPva had antimicrobial activities and/or interacted with type 1 fimbriae, respectively. Finally, bacterial adherence and invasion to cells treated with LPva was examined.

Results

LPva induced uroepithelial apoptosis which was coupled with upregulated expression of caveolin-1 and downregulation of β1 integrin. LPva did not exhibit direct antimicrobial properties and did not influence antimicrobial peptide levels in cells. Additionally, LPva did not interact with type 1 fimbriae and did not affect adherence in comparison to non-treated control cells. However, LPva significantly reduced the number of intracellular UPEC in bladder epithelial cells.

Conclusions

Our findings suggest that LPva has beneficial applications against UPEC infection due to its ability to induce programmed cell death and reduce bacterial invasion of the uroepithelium.     

A protective effect of Curcuma comosa Roxb. on bone loss in estrogen deficient mice

Ethnopharmacological relevance

Curcuma comosa Roxb. or Wan chak motluk is an indigenous medicinal herb and has traditionally been used among postmenopausal women for relief of unpleasant menopausal symptoms.

Aim of the study

Estrogen deficiency is a causative factor in the development of osteoporosis in menopausal women. Phytoestrogens, non-steroidal plant-derived compounds which have an array of beneficial effects, are considered as an effective alternative compound in preventing bone loss caused by the deficiency of estrogen. The present study determined the potential effect of Curcuma comosa Roxb. (C. comosa) hexane extract containing phytoestrogens in protecting bone loss induced by ovariectomy in mice.

Materials and methods

Mature Swiss albino female mice were ovariectomized and treated with the C. comosa extract for 5 weeks. Bone calcium content, bone mass density, histology, and bone markers were evaluated.

Results

The ovariectomized mice showed a marked decrease in total bone calcium content and bone mass density of lumbar vertebrae 5-6, femur and tibia bone in comparison with the intact control mice. Bone histology demonstrated the poor development of endochondral bone formation in ovariectomized mice which correlated with a decrease in plasma bone alkaline phosphatase activity. Treatment with C. comosa protected against the loss of total bone calcium content and bone mass density in both trabecular and cortical bones, similar to results observed with estrogen treatment. In addition, C. comosa treatment resulted in less increase in uterine weight compared to estrogen treatment.

Conclusion

Our results suggest that C. comosa prevents bone loss induced by estrogen deficiency. Therefore, C. comosa would be a potential alternative treatment for prevention of postmenopausal osteoporosis.     

In vivo anti-inflammatory activities of leaf extracts of Ocimum lamiifolium in mice model

Aim of the study

Ocimum lamiifolium Hochst. ex Benth. (Lamiaceae) has been used in Ethiopian traditional medicine for the treatment of different inflammatory disorders such as oropharyngitis, wound, pain, fever, and others. However, its use has mainly been based on empirical findings. Thus the objective of this study was therefore to evaluate the antiinflammatory effects and acute oral toxicities of the leaf extracts of Ocimum lamiifolium in mice.

Materials and methods

Aqueous and ethanol crude extracts were screened for their antiinflammatory activities in mice using carrageenin induced paw edema. And then the aqueous extract, the most active extract, was further fractionated and the fractions were tested for their anti-inflammatory activities using carrageenin, histamine and serotonin induced mice paw edema. Distilled water and aspirin were employed as negative and positive controls, respectively. Acute oral toxicity of both extracts and fractions were also determined after giving graded doses.

Results

The aqueous and ethanol extracts were able to reduce inflammation significantly, but greater anti-inflammatory activity was observed for the aqueous extract at all dose levels. Of all fractions the water residue showed highly significant anti-inflammatory activity.

Conclusions

Ocimum lamiifolium leaf extracts exhibited significant anti-inflammatory activities with less acute toxicity.