The impact of Zataria multiflora Boiss extract on in vitro and in vivo Th1/Th2 cytokine (IFN-γ/IL4) balance

Ethnopharmacological relevance

Anti-inflammatory, anti-oxidant and effect of Zataria multiflora on Th₁/Th₂ balance were previously described. Different therapeutic effects of this plant have been described in Iranian traditional medicine. To evaluate the immune modulatory effects of Zataria multiflora on Th1/Th2 balance, which may be implicated in inflammatory disorders, in vitro and in vivo studies were carried out.

Materials and methods

The effects of three concentrations of the extract, dexamethasone, and saline on interleukin 4 (IL-4) and interferon γ (IFN-γ) gene expression were evaluated in phytohemagglutinin (PHA) stimulated and non-stimulated human peripheral blood mononuclear cells (hPBMCs). RNA was extracted from the hPBMCs to make cDNA for real time PCR relative quantification. Furthermore, the effect of the extract on serum level of IL-4 and IFN-γ was assessed in ovalbumin (OA) sensitized guinea pigs (n=6 for each group).

Results

Dexamethasone showed significant inhibitory effect on both IFN-γ and IL-4 gene expression and serum level of the cytokines and significantly enhanced IFN-γ/IL-4 ratio (p<0.05–p<0.001). The extract inhibited IL-4 and enhance IFN-γ gene expression and IFN-γ/IL-4 ratio too (p<0.05–p<0.001). In sensitized animals also serum level of IL-4 were significantly decreased after treatment with both dexamethasone and extract, but serum level of IFN-γ and IFN-γ/IL-4 ratio were significantly increased due to extract treatment (p<0.01 for medium and p<0.001 for high concentration).

Conclusions

These results indicated consistent in vitro and in vivo data for selective immune modulatory effect of the extract of Zataria multiflora which increased IFN-γ, decreased IL-4, and enhanced the ratio of IFN-γ to IL-4 (Th₁/Th₂ balance). Therefore, the extract of Zataria multiflora may have therapeutic value in inflammatory responses such as allergy, autoimmunity and infectious diseases associated with Th₁/Th₂ imbalance.     

Hoodia gordonii: a natural appetite suppressant

Aims of the study

Several species of the stapeliads, a group of stem succulents belonging to the family Apocynaceae are reported on in the ethnopharmacology literature and many of the references relate to their use as food plants. The most important of these plants is Hoodia gordonii, which during the past decade has risen from an almost forgotten spiny, desert plant to an important commercial appetite-suppressant herbal. The aim of this review is to summarize the botany, ethnopharmacology and phytochemistry of Hoodia gordonii.

Materials and methods

Journal articles and books were used to collect information on Hoodia gordonii and related species.

Results

Many books and articles documented the use of stapeliad species as food plants and earlier references refer to the use Hoodia species as a thirst quencher. However, prior to the publication of the patent application, only a single reference referring to the use of Hoodia pilifera as appetite suppressant was found. The structures of several steroid glycosides isolated from Hoodia gordonii are summarized.

Conclusions

Hoodia gordonii illustrates how a combination of ethnobotany and scientific research can lead to a commercial product which can greatly benefit the indigenous people.     

Calotropis gigantiea (L.) R. Br (Apocynaceae): A phytochemical and pharmacological review

Ethnopharmacological relevance

Calotropis gigantiea (L.) R. Br (Apocynaceae) commonly called as “crown flower” or “giant milk weed” is a well-known weed to many cultures for treating various disorders related to central nervous system, skin diseases, digestive system, respiratory system, reproductive system etc. Indigenous groups made the plant as a part of their lives since they use the fruit fibre to make ropes, household items, for weaving clothes and flowers for garlands apart from usage for various indications. The study aims at far-reaching review on phytochemistry, pharmacological activities, ethnopharmacology, intellectual property transfer on pharmacological therapies, toxicity which aids to provide scientific evidence for the ethnobotanical claims and to identify gaps required to be conducted as a future research prerequisite.

Materials and methods

A systematic literature search was performed using different databases such as Scopus, Science direct, PubMed and Sciverse with no timeline limit set during the search. All the available abstracts and full text articles were included in the systematic review.

Results

Most of the folkloric uses were validated by the scientific studies such as analgesic, anti-arthritic, anti-asthmatic, anti-bacterial, anti-convulsant, anti-pyretic, central nervous system disorders, contraceptive, anti-ulcer and wound healing. In addition other studies such as anti-diabetic, anti-diarrhoeal, anti-helminthic, anti-histamine, anti-inflammatory, anti-microbial, anti-oxidant, cardio-protective studies, cytotoxicity, hepatoprotectivity, fibrinolytic, mosquitocidal, nerve muscle activity, vasodilation and skeletal muscle activities were also reported for the plant. Isolated compounds such as calotropin, frugoside and 4'-O-β-d-glucopyranosyl frugoside were tested for the cytotoxicity efficacy against both human and rat cell lines out of which calotropin showed potent activity (IC₅₀–15 ng/ml). However there were no clinical trials reported on the plant which is one of the major lacunas.

Conclusions

This review article explores the ethnopharmacological, pharmacological activities phytochemistry and intellectual rights of Cg which gives the evidence of a potent and commercial drug which up on further research leads to the most viable drug for variety of treatments. However there is further need for in-vivo studies and clinical trials on isolated phytoconstituents which will help to commercialise.     

Acute diuretic activity of aqueous Erica multiflora flowers and Cynodon dactylon rhizomes extracts in rats

Aim of the study

The aim of the present study is to evaluate the diuretic potential and effect on urinary electrolytes of aqueous Erica multiflora L. (Ericaceae) flowers and Cynodon dactylon L. (Poaceae) rhizomes extracts in rats.

Materials and methods

Different concentrations of these plants extract (0.125, 0.250, and 0.500 g/kg of body weight) or the reference drug furosemide (0.015 g/kg) were administrated orally to hydrated male Wistar rats and their urine output was measured at several interval of time after a single dose administration. Furthermore, a toxicological effect of both plants was undertaken as well.

Results

The results showed that furosemide induced significant diuresis and electrolytes excretion during the first hours. Plant extracts increased significantly urinary output and electrolytes excretion at the dose of 0.250 g/kg for Erica multiflora and 0.500 g/kg for Cynodon dactylon. This diuretic effect seems to be not related to K⁺ plant content. Urinary pH remained mostly unchanged during the course of the study for both plant extracts. No lethality was observed among animals when using Erica multiflora even at the dose of 10 g/kg while Cynodon dactylon, instead, caused 50% of rat death (LD50) at 4.5 g/kg.

Conclusion

We concluded that both aqueous herb extracts administered, particularly, at the dose of 0.500 g/kg induce significant effect on urinary output of water and electrolytes and justify their use as diuretic remedy in traditional medicine.     

Studien zum Lingshu als Beitrag zur Bewertung klinischer Akupunkturstudien: – Punktlokalisation,Stichtiefe, De Qi

Background

Clinical trials on acupuncture use a variety of control methods, such as sham acupuncture, minimal acupuncture, and placebo. Frequently, needle insertion on non-classical acupuncture points (“minimal acupuncture”) is employed for this purpose. The aim of such clinical trials is to evaluate specific effects of acupuncture. There is a controversial discussion regarding the appropriateness of comparing “true” acupuncture to “minimal acupuncture”.Minimal and verum acupuncture are distinguished by various criteria, such as “localisation”, “depth of needle insertion”, and “induction of de qi”.

Aim

This study is dedicated to the question of what information the Huang Di Neijing, specifically the Lingshu-compilation which extensively focuses on needling techniques, has to offer regarding the above mentioned criteria, and their importance for successful acupuncture. In relation to this, it will be discussed if the definition of “minimal acupuncture” as used in the German Acupuncture Trials, is feasible.

Methods

The entire original text of the Huang Di Neijing Lingshu was searched for information regarding “point localisation”, “insertion depth”, and “induction of “de qi”. The paragraphs in question were compiled and probed on statements regarding the efficacy of treatment. These subsequently were compared to the technique of “minimal acupuncture” as in the German Acupuncture Trials (ART, GERAC).

Results

The analysis of the Lingshu shows that neither a specific point localisation, nor a certain insertion depth or the induction of de qi are clearly and in all cases relevant for a successful therapy. Rather, the Lingshu offers specific indications for superficial needling. Sites of acupoint localisation aren't clearly defined and the induction of de qi is not understood as mandatory for a successful therapy.

Conclusion

The criteria distinguishing „verum acupuncture” from „minimal acupuncture” as practiced in many clinical trials don't correspond to the early practice of acupuncture as documented in the Lingshu. This can be a possible explanation for the good clinical results of minimal acupuncture in a number of clinical trials.However, for further discussions of this problem the developments and the clinical evolution present-day acupuncture has undergone since the ancient times have to be held in account.     

Clerodendrum serratum (L.) Moon. – A review on traditional uses,phytochemistry and pharmacological activities

Ethnopharmacological relevance

Clerodendrum serratum (L.) Moon. (Verbenaceae) is an important medicinal plant growing in the tropical and warm temperate regions like Africa, Southern Asia; Malaysia and distributed throughout in forests of India and Sri Lanka. It is traditionally valued and reported for treating pain, inflammation, rheumatism, respiratory disorders, fever and malarial fever in India with a long history. To provide a comprehensive overview of the traditional and ethno medicinal uses, phytochemistry and biological activities of C. serratum with clinical and toxicity data and possibly make recommendations for further research.

Materials and methods

All relevant worldwide accepted databases were searched for the terms “Clerodendrum”, “Clerodendrum serratum”, “Bharangi” and “Cheruthekku” along with the other literature from Indian classical texts and pharmacopoeias. There was no specific timeline set for the search. The accessible literatures available on C. serratum were collected via electronic search using Pubmed, Scopus, Science Direct and traditional books reports on ethnopharmacology and traditional medicines.

Results

C. serratum has played an important role in Indian system of medicine. In addition to the common local use in respiratory diseases, other ethnomedicinal uses include treatment of pain, inflammation, rheumatism and fever especially malarial fever. Scientific studies on extracts and formulations revealed anti-asthmatic, mast cell stabilization and anti-allergic effects of roots of C. serratum. Reported data on pharmacological activities also includes hepatoprotective, anti-oxidant, anti-inflammatory and anticancer potential of the drug. Saponins (terpenoids and steroids), flavonoids and phenolics isolated from roots have been the focus of phytochemical investigations as the biological activity has been ascribed to the saponins, which are known to possess anti-inflammatory and anti-cancer activity. Isolated bioactives from roots like icosahydropicenic acid and ursolic acid have been claimed to offer anti-allergic and hepatoprotective activity.

Conclusions

Therapeutic potential of roots and leaves of C. serratum has been demonstrated in the conditions like asthma, allergy, fever, inflammation and liver disorders attributed to the presence of various flavonoids, phenolics and saponins present in the drug. Many ethnobotanical claims have been confirmed through modern in-vitro and in-vivo pharmacological studies of different extracts and isolates from plant; however, additional studies on the biomarkers are needed to establish mechanism of action and to validate the traditional use of this drug in clinical practices after proper safety assessment.     

Characterization of the antinociceptive and anti-inflammatory activities from Cocos nucifera L. (Palmae)

Aim of the study

Cocos nucifera cultivated in Brazil is known as “coco-da-Bahia” or “coqueiro-da-Índia”. The tea from the husk fiber is widely used to several inflammatory disorders. Crude extract and fractions obtained from Cocos nucifera “common variety” were evaluated to test the anti-inflammatory and antinociceptive activities.

Materials and methods

Crude extract (CE, 50, 100, and 150 mg/kg), fraction 1 (F1, molecular weight lesser than 1 kDa, 1, 10, and 50 mg/kg), fraction 2 (F2, molecular weight higher than 1 kDa, 1, 10, and 50 mg/kg), and the references drugs morphine (5 mg/kg), acetilsalicilic acid (200 mg/kg), prometazine (30 mg/kg), and metisergide (5 mg/kg) were evaluated on models of analgesia and inflammation.

Results

CE, F1, and F2 significantly develop peripheral and central antinociceptive activity but with less effect on supra-spinal regions of the brain. Administration of the opioid antagonist, naloxone (5 mg/kg) inhibited the antinociceptive effect indicating that Cocos nucifera crude extract and fractions may be acting in opioid receptors. CE and F1 also inhibited rat paw edema induced by histamine, and serotonin.

Conclusions

results demonstrated that Cocos nucifera and its fractions have antinociceptive and anti-inflammatory activities which confirm the popular use of this plant in several inflammatory disorders.     

Genotoxic and antigenotoxic properties of Baccharis trimera in mice

Ethnopharmacological relevance

Baccharis trimera (Less.) (Asteraceae) is a native plant from Brazil. Also known as “carqueja”, it is popularly used to treat liver diseases, diabetes, as well as digestive disorders, mainly by women with lower socioeconomic status.

Aim of the study

The aim of the present study was to investigate the in vivo genotoxic/antigenotoxic and mutagenic potential of this plant, using the comet and the micronucleus assays.

Material and methods

Female adult mice were treated with 500, 1000 or 2000 mg/kg of B. trimera aqueous extract (Bt-AE) by gavage for three consecutive days.

Results

Independently of the dose, no genotoxic effect was observed in blood and liver samples analyzed by the comet assay. Conversely, B. trimera showed an antigenotoxic effect in blood from treated mice, thus protecting cells against oxidative DNA damage induced by the ex vivo treatment with hydrogen peroxide. In addition, Bt-AE showed in vitro antioxidant activity, assessed by DPPH and xanthine oxidase assays, suggesting that the observed antigenotoxic effects might be related to its antioxidant properties.

Conclusions

However, the extract increased the frequency of micronucleus in bone marrow of treated mice, indicating a chromosomal mutagenic activity. Thus, medicines prepared from this plant should be used with caution, although the results also suggest antigenotoxic effects for B. trimera aqueous extract.     

An herbal decoction of Radix astragali and Radix angelicae sinensis promotes hematopoiesis and thrombopoiesis

Ethnopharmacological relevance

A decoction containing Radix angelicae sinensis and Radix astragali (Danggui Buxue Tang, DBT) has been used to raise the “Qi” and nourish the “Blood”. However, its effects on haematopoiesis and particularly thrombopoiesis have not been studied.

Aims

This study aims to examine the effects of DBT on hematopoiesis and thrombopoiesis.

Materials and methods

A myelosuppression mouse model was treated with DBT (10 mg/kg/day). Peripheral blood cells from DBT and thrombopoietin-treated samples were counted on days 0, 7, 14 and 21. Then CFU assays were used to determine the effects of DBT on the megakaryocytic progenitor cells and other lineages. Last, analyses of annexin V, caspase-3, and mitochondrial membrane potential were conducted in megakaryocytic cell line M-07e.

Results

Morphological examination showed that DBT treatment significantly increased the recovery of the megakaryocytic series. DBT significantly enhanced the platelet recovery and CFU-MK formation in vivo. DBT significantly promoted CFU-MK and CFU-F formation. Last, we observed the antiapoptotic effects of DBT on M-07e cells.

Conclusion

DBT might promote haematopoiesis and thrombopoiesis in the mouse model through (i) directly promoting the growth of megakaryocytes; (ii) indirectly promoting the growth of bone marrow stromal cells; (iii) inhibiting apoptosis of megakaryocytes.     

Assessment of anti-protozoal activity of plants traditionally used in Ecuador in the treatment of leishmaniasis

Aim of the Study

For the assessment of the in vitro anti-protozoal potential of plants traditionally used in Ecuador in the treatment of leishmaniasis, a combined approach based on interviews with healers as well as a literature search was carried out.

Materials and Methods

From three regions of Ecuador, 256 local healers called “Agents of Traditional Medicine” (ATMs) were interviewed about their knowledge of the use of plants to treat and heal the illness recognized by the ATMs as leishmaniasis. From literature sources, 14 plants were identified as being used in the treatment of leishmaniasis. Subsequently, plant material was collected from a representative selection of 39 species. A total of 140 extracts were screened in vitro against Leishmania donovani, Plasmodium falciparum, Trypanosoma brucei rhodesiense and Trypanosoma cruzi. Additionally, these extracts were evaluated for their anti-microbial activities using five gram-positive and -negative bacteria as well as Candida albicans.

Results and Conclusions

The survey resulted in 431 use-records for 145 plant-taxa used for the treatment of leishmaniasis. The 10 most frequently reported taxa accounted for 37.7% of all records. In the case of leishmaniasis, activity was observed for Elephantopus mollis, Minquartia guianensis, Bocconia integrifolia, Gouania lupuloides, Scoparia dulcis, an as-yet-unidentified species of Piper and Brugmansia. For the leaves of M. guianensis and the twigs and bark of G. lupuloides a good selectivity index (SI) was found. IC₅₀ values and the SI of active plant extracts are presented.     

Involvement of glutamate and cytokine pathways on antinociceptive effect of Pfaffia glomerata in mice

Ethnopharmacological relevance

Pfaffia glomerata (Spreng) Pedersen (Amaranthaceae) is a medicinal plant known in Brazil as “Paratudo” and “Brazilian ginseng” and is commonly used as tonic, antidiabetic and to treat gastric disorders.

Aim of the study

This study evaluates the possible mechanism by which hydroalcoholic extract (HE) of Pfaffia glomerata exerts its antinociceptive effect.

Materials and methods

The HE was evaluated in acetic acid and glutamate models of pain or by biting behavior following intrathecal (i.t.) administration of agonists of excitatory aminoacids (EAA) receptors glutamate and pro-inflammatory cytokines, IL-1β and TNF-α in mice.

Results

Oral administration of HE produced dose-dependent inhibition of acetic acid-induced visceral pain and glutamate-induced pain, with ID₅₀ of 64.6 (47.7–87.5) mg/kg and ID₅₀ of 370.8 (253.4–542.7) mg/kg, respectively. The HE (300 mg/kg, p.o.) antinociception, in the acetic acid test, was not affected by i.p. treatment of animals with naloxone. In addition, HE (300 mg/kg, p.o.) inhibited the pain-related behaviors induced by i.t. injection of trans-ACPD and TNF-α, but not by NMDA, AMPA, kainate or IL-1β.

Conclusions

Our results suggest that inhibition of glutamatergic metabotropic receptors and TNF-α may account for the antinociceptive action reported for the HE in models of chemical pain used in this study.     

Central nervous system effects and chemical composition of two subspecies of Agastache mexicana; an ethnomedicine of Mexico

Ethnopharmacological relevance

Agastache mexicana subspecies mexicana (Amm) and xolocotziana (Amx) are used in Mexican traditional medicine to relief cultural affiliation syndromes known as “susto” or “espanto”, for “nervous” condition, and as a sleep aid. Despite its intensive use, neuropharmacological studies are scarce, and the chemical composition of the aqueous extracts has not been described. Aims of the study are: (1) To analyze the chemical composition of aqueous extracts from aerial parts of Amm and Amx. (2) To evaluate the anxiolytic-like, sedative, antidepressant-like effects. (3) Analyze the general toxic effects of different doses.

Materials and methods

Anxiolytic-like and sedative effects were measured in the avoidance exploratory behavior, burying behavior and the hole-board tests. The antidepressant-like actions were studied in the forced swimming and tail suspension tests. Finally, general activity and motor coordination disturbances were evaluated in the open field, inverted screen and rota-rod tests. The acute toxicity of Amm and Amx was determined by calculating their LD₅₀ (mean lethal dose). The chemical analyses were performed employing chromatographic, photometric and HPLC–ESI-MS techniques.

Results

Low doses of Amm and Amx (0.1σ1.0 mg/kg) induced anxiolytic-like actions; while higher doses (over 10 mg/kg) induced sedation and reduced the locomotor activity, exerting a general inhibition in the central nervous system (CNS).

Conclusions

Results support the use of Amm and Amx in traditional medicine as tranquilizers and sleep inducers. Additionally, this paper contributes to the knowledge of the chemical composition of the aqueous extracts of these plants.     

Anti-inflammatory compounds of “Qin-Jiao”, the roots of Gentiana dahurica (Gentianaceae)

Ethnopharmacological relevance

“Qin-Jiao” is a well-known traditional Chinese medicinal (TCM) herb having been used generally for fighting rheumatoid arthritis (RA) since ancient times. The root of Gentiana dahurica Fisch (Gentianaceae) is one of the four officially validated “Qin-Jiao” as listed in the Chinese Pharmacopoeia. In addition, it is a common Tibetan medicinal herb used for the treatment of tonsillitis, urticaria, and RA, while the flowers have been used as a Mongolian herb for curing cough sore throat and eliminating the phlegm due to its anti-inflammatory effect.

Aim of the study

The aim of the study was to characterize the anti-inflammatory compounds in “Qin-Jiao”, on the basis of detailed investigation on not only the phytochemical study of Gentiana dahurica, but also the bioactive evaluation on compounds obtained presently and previously from different “Qin-Jiao” origins and Gentiana species.

Materials and methods

The ethanol extract of air-dried roots of Gentiana dahurica was suspended into H₂O and extracted with EtOAc and n-BuOH, successively. Repeated column chromatography (CC) and semi-preparative HPLC were carried out on each of the fractions. The isolated compounds were determined by detailed spectroscopic analysis and acidic hydrolysis. Anti-inflammatory activities of 18 isolates, together with 12 typical compounds obtained previously by our group from the other “Qin-Jiao” origins (Gentiana crassicaulis, Gentiana straminea) and Gentiana rigescens, were tested by inhibitory effects on LPS-induced NO production in macrophage RAW264.7 cells and TPA-induced cyclooxygenases-2 and -1 (COXs-2/1) production on zebrafish model.

Results

A new lignan glycoside (1) was identified, together with 20 known compounds, including 10 iridoid glycosides (2–11), three steroids (12–14), four lignans (15–18), one phenylpropanoid (19) and two triterpenes (20–21). Anti-inflammatory bioassay showed that only compound 21 displayed potential inhibitory effect on NO production (IC₅₀=16.85 μM), while 20 tested compounds had inhibitory activities on COXs-2/1. Among them, the triterpenoid 21 was the most active compound with an inhibitory value of 78% at a concentration of 30 μM. All the tested compounds showed no cytotoxicity on five human cancer cell lines (40 μM) and zebrafish (30 μM), except for 21 displaying weak cytotoxicity on human myeloid leukemia HL-60 (IC₅₀=16.43 μM).

Conclusion

Most of compounds particularly iridoid glycosides from “Qin-Jiao” display potential inhibitory effect on COXs-2/1. The results support the historical importance of the well-known TCM herb, “Qin-Jiao”, having been commonly used for fighting RA. As major components, the bioactive iridoid glycosides should play important role in the anti-inflammatory effect of “Qin-Jiao”. Although further research will be required to evaluate the selective activities of the COXs-2/1 inhibitors, this work validates the medicinal use of “Qin-Jiao” and provides information for different “Qin-Jiao” origins having different treating effects on RA.     

Antinociceptive effect of ethanol extract of leaves of Lannea coromandelica

Ethnopharmacological relevance

Lannea coromandelica (Houtt.) Merr. is a plant locally called “Jiga”, found all over Bangladesh. Leaf of the plant is traditionally used in the treatment of local swellings, pains of body, toothache etc. This study evaluated the antinociceptive effect of the ethanol extract of L. coromandelica leaves (EELC).

Materials and methods

The antinociceptive activity of the extract (at the doses of 50, 100, and 200 mg/kg) was evaluated by using chemical- and heat-induced pain models such as acetic acid-induced writhing, hot plate, tail immersion, formalin, and glutamate test. To verify the possible involvement of opioid receptor in the central antinociceptive effect of EELC, naloxone was used to antagonize the effect. Besides, the involvements of ATP-sensitive K⁺ channel and cGMP pathway were also justified by using glibenclemide and methylene blue.

Results

EELC demonstrated significant dose-dependent antinociceptive activity in the chemical- and heat-induced nociception in mice models (p<0.05). These findings imply the involvement of both peripheral and central antinociceptive mechanisms. The use of naloxone confirmed the association of opioid receptors in the central antinociceptive effect. EELC also showed the involvements of ATP-sensitive K⁺ channel and cGMP pathway for antinociceptive activity.

Conclusions

This study reported the antinociceptive activity of the leaf of L. coromandelica and rationalized the traditional use of the leaf in the treatment of different painful conditions.     

Novel bisabolane derivative from “arnica-da-serra” (Vernonieae: Asteraceae) reduces pro-nociceptive cytokines levels in LPS-stimulated rat macrophages

Ethnopharmacology relevance

Hydro alcoholic leaves extracts (HALE) of Lychnophora ericoides Mart. (“false arnica” or “arnica-da-serra”) had been popularly used against pain and inflammatory process.

Aim

The present work aimed to look for possible active volatile compounds that could be found in HALE of Lychnophora ericoides among the non volatile anti-inflammatory and analgesic compounds previously reported.

Methods

Harvests were performed during the end of the wet summer season (April) when scented branches were instantly collected and frozen. HALE's were simulated at the lab by following the procedures lectured by the locals. Mass Spectrometry experiments suggested structural information when using both EI–MS and ESI–MS/MS. After isolation through classical thin layer chromatography (TLC) procedures, the NMR experiments and signals assignments were carried out. The effects on the cytokines or nitric oxide (NO) production were assessed at in vitro assays that had monitored the levels of these substances on the supernatant of LPS-stimulated macrophage primary cell culture.

Results

The major metabolite from HALE was isolated from the essential oil and the major compound had its molecular formulae established by Mass Spectrometry (High Resolution) and its structure by NMR. Literature-based investigation enables us to define the structure of the new metabolite as 6-methyl-2-(4-methylcyclohex-4-enyl-2-acetyloxy) hept-5-en-2-ol and its name as orto-acetoxy-bisabolol. In vitro assay of interleukins release inhibition was carried out using rat peritoneal macrophages cultures. IL-1β and TNF-α levels were significantly reduced when cells were previously treated with low doses of orto-acetoxy-bisabolol, but neither IL-6 nor NO levels have their levels reduced. Results suggest that ethnical knowledge of anti-inflammatory and analgesic effects of the “arnica-da-serra” HALE may be associated to the orto-acetoxy-bisabolol ability on synthesis inhibition of the key inflammatory/hypernociceptive mediators.

Conclusions

Phytochemical investigation of the volatile active compounds in Lychnophora ericoides HALE allows us to isolate a new bisabolane derivative (orto-acetoxy-bisabolol) and to infer that this compound inhibits the synthesis of TNF-α and IL-1β, two important inflammatory mediators in the hypernociception. Our present data, in addition to literature's data, furnish scientific support to folk's use of Lychnophora ericoides as an endemic wound healer.     

The future is written: impact of scripts on the cognition, selection, knowledge and transmission of medicinal plant use and its implications for ethnobotany and ethnopharmacology

Aim of the study

Apart from empirically learned medicinal and pharmacological properties, the selection of medicinal plants is dependent on cognitive features, ecological factors and cultural history. In literate societies the transmission of medicinal plant knowledge through texts and, more recently, other media containing local as well as non-local knowledge has a more immediate and a more prolonged effect than oral transmission. Therefore, I try to visualize how field based studies in ethnobiology and especially medical ethnobotany and ethnopharmacology run the risk of repeating information and knowledge and illustrate the importance of differentiating and acknowledging the origin, transmission and rationale of plant use made by humans.

Materials and methods

Reviewing literature dealing with the traditional parameters (e.g. hot/cold dichotomy, organoleptic properties, doctrine of signatures) influencing the selection and transmission of plant use in a juxtaposition to our recent finding of causal influence of text on local plant use. Discussing the passing down of knowledge by text as a special case of oblique/one-to-many knowledge transmission.

Results

Historical texts on materia medica, popular books on plant use, clinical studies, and informants of ethnobotanical field studies generate a circle of information and knowledge, which progressively conditions the results of ethnobotanical field studies. While text reporting on phytotherapeutical trends may cause innovation through the introduction of “new” applications to local customs, persistently repeating well established folk remedies leads to the consolidation of such uses adding a conservative dimension to a local pharmacopoeia, which might not actually be there to that extent.

Conclusions

Such a “shaping” of what might appear to be the results of a field investigation is clearly outside the ordinary principles of scientific enquiry. The traditional pillars of ethnobotanical field studies - that is, “input to drug discovery” and “conservation of cultural heritage” - are also incompatible with this process. Ethnobotancial field studies aimed at a contribution to natural products research and/or the conservation of cultural heritage, as well as those aimed at an assessment and validation of local pharmacopoeias should differentiate between local plant use and widespread as well as modern knowledge reported in popular textbooks and scientific literature.     

Ameliorative and neuroprotective effect in MPTP model of Parkinson's disease by Zhen-Wu-Tang (ZWT), a traditional Chinese medicine

Aim of the study

Traditional Chinese medicine Zhen-Wu-Tang (ZWT) is a well-known PentaHerbs formula from “Treatise on Febrile Disease”. This study is to elucidate its neuroprotective effect and mechanism of ameliorative effect of the syndrome of Parkinson's disease (PD).

Materials and methods

The ameliorative effect of ZWT on symptom of PD through behavior tests including: swimming test, the tail suspension test and open-field test was investigated. The neuroprotective effect of dopaminergic neurons from the striatum and frontal cortex of brain was detected by high performance liquid chromatography with electrochemical detection (HPLC-ECD).

Results

This study proved that ZWT could ameliorate the typical symptom of PD and protect dopaminergic system.

Conclusion

These results suggested that ZWT possessed protective and ameliorative properties of dopaminergic neurons.     

Chemical composition,potential toxicity,and quality control procedures of the crude drug of Cyrtopodium macrobulbon

Ethnopharmacological relevance

Cyrtopodium macrobulbon (“cañaveral”) has been long used in Mexican traditional medicine for the treatment of painful urinary ailments (“mal de orin”) in men. This study was conducted (i) to establish the potential acute toxicity and the antinociceptive activity of some preparations of Cyrtopodium macrobulbon, in order to demonstrate its preclinical efficacy for treating symptoms of “mal de orin”; and (ii) to determine the chemical composition and quality control parameters of this medicinal orchid.

Materials and methods

The antinociceptive effect was assessed using the acetic acid-induced writhing and the hot-plate tests. Investigation of the acute toxicity was accomplished by the Lorke method. The organic extract (OE) was subjected to conventional phytochemical study using chromatographic conventional procedures. The volatile components profile of the species was accomplished via GC–MS analysis of HS-SPME-adsorbed compounds. Furthermore, an HPLC method to quantify ephemeranthol B (10) was developed and validated according to the International Conference on Harmonization Guidelines. Microscopic anatomy studies were performed using light and scanning electron microscopies. Finally, a potential distribution map was generated using the MaxEnt modeling method.

Results

AE and OE were not toxic to mice since the LD₅₀ was higher than 5000 mg/kg. OE was only active in the acetic acid-induced writhing assay at the doses of 100 and 316 mg/kg. Conventional phytochemical analysis of OE led to the isolation and characterization of n-hexacosyl-trans-p-coumarate (1), n-octacosyl-trans-p-coumarate (2), n-triacontyl-trans-p-coumarate (3), 4-methoxy-benzyl alcohol (4), 4-hydroxybenzaldehyde (5), 1,5,7-trimethoxy-9,10-dihydrophenanthrene-2,6-diol (6), confusarin (7), gigantol (8), batatasin III (9), and ephemeranthol B (10). The major volatile components identified by HS-SPME analysis were 6,10,14-trimethyl-2-pentadecanone, eucalyptol (11), and isobornyl formate. An HPLC analytical method for the quantification of compound 10 in the plant was developed and fully validated for selectivity, accuracy, and precision. The microscopic studies revealed that the epidermal tissue displayed a layer of enlarged, crenate and cell thin-walled cells with a thickened cuticle; these cells are described for first time for this species. The potential distribution map generated revealed that this species is widespread in Mexico from Sinaloa to Merida states.

Conclusions

The results of the pharmacological studies tend to support the traditional use of Cyrtopodium macrobulbon for “mal de orin”; the presence of compounds 8, 9, and 11 with known antinociceptive activity might be related with the pharmacological effect demonstrated. The HPLC and microscopic analyses developed in this work will be valuable tools for quality control purposes for this plant.