The developmental neurobehavioral effects of fenugreek seeds on prenatally exposed mice

Ethnopharmacological relevance

Fenugreek (Trigonella foenum graecum (L.)), is a medicinal plant whose seeds and leaves are widely used in Moroccan traditional medicine. Consumption of fenugreek seeds during pregnancy has been associated with a range of congenital malformations, including hydrocephalus, anencephaly and spina bifida.In previous work we have shown that exposure of pregnant mice to Aqueous Extract of Fenugreek Seeds (AEFS) leads to reduced litter size, intrauterine growth retardation, and malformations. However, there have been no studies to date of its longer-term neurobehavioral effects. We investigated these effects in prenatally exposed mice.

Materials and methods

Pregnant females were exposed to 0, 500 or 1000 mg/kg/day AEFS, by gavage, for the whole period of gestation. Pups body weight was measured at 1, 7, 14, 21 and 28 day of age. Behavior of progeny was evaluated three weeks after birth using the open field, the rotarod test and the continuous alternation task by the T-maze. At 28 postnatal day age, brain of progeny was removed and cut for histological evaluation.

Results

The progeny of exposed mice displayed reduced body weight at birth (1000 mg/kg group: 27%; 500 mg/kg group: 32%) and reduced brain weight (10% in both treated groups). Both males and females mice prenatally exposed to AEFS displayed a significant decrease in the locomotor activity, in the boli deposits during the open field test and in motor coordination. These results seem to show that exposure to AEFS induces a depressive effect in the offspring. Assessment on a continuous alternation T-maze test showed a significant reduction in successful spontaneous alternations in males and females but only in the 1000 mg/kg group.

Conclusion

These results suggest that prenatal exposure of mice to high dose of fenugreek seeds causes growth retardation and altered neurobehavioral performance in the post-weaning period in both male and female.     

Role of Syzygium cumini seed extract in the chemoprevention of in vivo genomic damage and oxidative stress

Ethanopharmacological relevance

The seeds of Syzygium cumini, Skeels (Jamun) are extensively used in India for treatment of diabetes and other ailments.

Aim of the study

The aim of this work was to assess the role of Jamun seed extract (JSE) as a chemoprotective agent against in vivo oxidative stress and genomic damage.

Materials and methods

Experiments were carried out to evaluate in vitro protective effects of JSE against hydroxyl radical induced damage in pBR322 DNA, and in vivo genomic damage and oxidative stress in mice which received JSE orally for 5 days before exposure to genotoxic carcinogens urethane (URE) and 7,12-dimethyl benz(a)anthracene (DMBA).

Results

Aqueous and ethanolic extracts of JSE showed significant protective effects against hydroxyl radical induced strand breaks in pBR322 DNA. The in vivo experiments with aqueous JSE showed significant protective effects against chromosomal damage induced by the genotoxic carcinogens URE and DMBA. Biochemical assays registered significant inhibition of hepatic lipid peroxidation and increase in GSH level and activity of GST, SOD and CAT.

Conclusion

Our findings suggest that JSE can possibly play an important role as a chemopreventive agent against oxidative stress and genomic damage.     

Practices of traditional birth attendants in Machakos District, kenya

Ethnopharmacological relevance

The aim of the study was to document TBAs practices as well as the indigenous herbal remedies they use to manage pre, intra and post partum complications in a rural Kenyan community.

Materials and Methods

A cross sectional study was conducted on practicing TBAs and their clients living in the study area. Data was collected using structured questionnaires and interviews. Focused group discussions were held with the TBAs to supplement the interviews and questionnaire survey.

Results

Two hundred TBAs and 20 clients were interviewed. The majority of the TBAs were females 75% of them having attended to over 200 pregnant women over a period of 5 years and above compared to only 6% of the males. A total of 10 pregnancy related complications and symptoms including threatened abortion, labor complications, post partum hemorrhage and retained after birth were recorded. Fifty five plant species most of them belonging to Euphorbiaceae family were identified for the management of the complications.

Conclusion

Traditional Birth Attendants still have a role to play in assisting pregnant women in rural communities. Their knowledge on herbal medicines is equally important and should be preserved for posterity.     

Polyphyllin D, a steroidal saponin from Paris polyphylla, inhibits endothelial cell functions in vitro and angiogenesis in zebrafish embryos in vivo

Ethnopharmacological relevance

Angiogenesis, the process of blood vessel formation, is critical to tumour growth. The importance of angiogenesis in tumour development has lead to the development of anti-angiogenic strategies to inhibit tumour growth. In this study, polyphyllin D (PD), an active component in Chinese herb, Paris polyphylla, was evaluated for its potential anti-angiogenic effects.

Materials and methods

The inhibitory effects of PD on three important processes involved in angiogenesis, i.e. proliferation, migration and differentiation were examined using human microvascular endothelial cell line HMEC-1 by MTT assay, scratch assay and tube formation assay, respectively. Using zebrafish embryos as an animal model of angiogenesis, the anti-angiogenic effect of PD was further verified in vivo.

Results

PD suppressed the growth of HMEC-1 cells at 0.1-0.4 μM without toxic effects. At 0.3 μM and 0.4 μM, PD significantly inhibited endothelial cell migration and capillary tube formation. About 70% of the zebrafish embryos showed defects in intersegmental vessel formation upon treatment with PD at concentrations of 0.156 μM and 0.313 μM.

Conclusion

The anti-angiogenic effects of PD have been explored in the study which implied a potential therapeutic development of PD in cancer treatment.     

Growth inhibitory and apoptosis inducing by effects of total flavonoids from Lysimachia clethroides Duby in human chronic myeloid leukemia K562 cells

Ethnopharmacological relevance

Lysimachia clethroides Duby is a traditional Chinese medicinal herb has been used in China to treat edema, jaundice diseases, hepatitis, tumor and inflammations, but the anti-tumor mechanisms are unclear.

Aim of the study

The present study was undertaken to investigate if total flavonoids from Lysimachia clethroides Duby (ZE4) possesses anti-cancer effects through apoptotic pathways in human chronic myeloid leukemia K562 cells.

Materials and methods

K562 cells were treated with different concentrations of ZE4 at different time intervals. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell apoptosis was assessed by flow cytometry, Hoechst 33258 staining and COMET assay. Western blot analysis was used to detect Bcl-2, Trail and DR5 expressions.

Results

15 flavonoids were isolated and identified from ZE4. ZE4 could inhibit the growth of K562 cells significantly by induction of apoptosis. Marked morphological changes of apoptosis, DNA fragmentation and single DNA strand breakages were observed clearly after treatment of ZE4. Bcl-2 expression was down-regulated remarkably while Fas, Trail and DR5 up-regulated when apoptosis occurred.

Conclusions

This result suggests that total flavonoids of Lysimachia clethroides Duby exert potential anti-cancer activity through growth inhibition and apoptosis in K562 cells.     

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.     

Alpha-momorcharin possessing high immunogenicity, immunotoxicity and hepatotoxicity in SD rats

Momordica charantia L., a genus of Momordica Linn. of the family Cucurbitaceae, commonly known as bitter melon, has been widely planted in China, Southeast Asia, Turkey and other areas, and has been used as a medicine for a long time. Alpha-momorcharin (α-MMC) extracted and purified from bitter melon seeds has significant anti-tumor and anti-virus effects, and has potential toxicity as well, especially when taken overdose. However, up to date studies on its safety evaluation are still insufficient.

Aims of the study

The immunogenicity, immunotoxicity and general toxicity of α-MMC were investigated in rats and guinea-pigs, and the potential toxic effects of the agent on the body were also examined.

Materials and methods

The major ribosome-inactivating protein was isolated by column chromatographies from the protein extracted from bitter melon seeds, and was verified as α-MMC. After rats were immunized by α-MMC, titers of specific antibody to α-MMC in immunized rats serum were detected by indirect ELISA. Guinea-pigs and rats immunized with α-MMC were used to evaluate the active systemic anaphylaxis and passive cutaneous anaphylaxis induced by α-MMC relatively. α-MMC of 6.25 mg/kg, 2.08 mg/kg and 0.70 mg/kg was administered to rats every 2 days. Five weeks later, animals were sacrificed, and then, biochemical examination, analysis of bone marrow and peripheral blood cells, and histopathologic examination were performed.

Results

The ribosome-inactivating protein isolated and purified from bitter melon seeds was identified as α-MMC. It induced high titer (1:46.4) of specific IgG and high positive results of the active systemic anaphylaxis and passive cutaneous anaphylaxis tests in animals. With the time of the α-MMC administration increasing, the body weights of the animals administered with α-MMC of 6.25 mg/kg decreased significantly, and point necrosis was also observed in liver cells, along with abnormal findings in serum chemistry, hematology and bone marrow histopathology test. The toxic effect lessened with the decrease of the dose of α-MMC and further reduced after the convalescence stage.

Conclusions

The results of the study show that α-MMC has high immunogenicity and immunotoxicity, and can cause obvious organic liver lesion.     

Biomarkers in the early period of acute myocardial infarction in rat serum and protective effects of Shexiang Baoxin Pill using a metabolomic method

Ethnopharmacological relevance

To identify the biomarkers in early period of acute myocardial infarction (AMI) in rat serum and reveal the effective mechanism of a Traditional Chinese Medicine (TCM) named Shexiang Baoxin Pill (SBP).

Material and method

A metabolomic approach using reversed-phase liquid chromatography/quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) was developed.

Results

Fourteen biomarkers in the early period of acute myocardial infarction (AMI) in rat serum were identified. These biomarkers include 5-methylcytosine, cystathionine ketimine, 2-oxoadipic acid, thymidine, epinephrine, homocystine, uric acid, 12(S)-hydroperoxyeicosatetraenoic acid (12s-HPETE), 11-dehydrocorticosterone, 12(S)-hydroxyeicosatetraenoic acid (12s-HETE), deoxycorticosterone, corticosterone, aldosterone and cortisol. Through pathway analysis of these biomarkers, inflammation, hypertrophy and oxidative injury were considered the most relevant pathological changes in early period of AMI.

Conclusion

Identification of AMI biomarkers not only supplied a systematic view of the progression of AMI in the early period but also provided the theoretical basis for the prevention or treatment of AMI. The results demonstrated that SBP pretreatment could offer protective effects for AMI through regulating the pathway of steroid hormone biosynthesis.     

Effects of Portulaca oleracea L. seeds in treatment of type-2 diabetes mellitus patients as adjunctive and alternative therapy

Ethnpharmacological relevance: To investigate antidiabetic activity of purslane seeds on type-2 diabetic subjects and to provide scientific basis for the clinical use of Portulaca oleracea (PO).

Materials and methods

A thirty subject with type-2 diabetes divided into two groups, to receive 5 g of PO seeds twice daily while in the second group, their participants receive 1500 mg of metformin/day. All participants were requested to report the effects of treatments on diabetic manifestations, their weights, body mass index (BMI), adverse effects, fasting and post-prandial blood glucose during treatment schedule. Blood samples from participants before and after treatment were taken for serum separation, which are used for measurement of serum lipids, liver enzymes, total and direct bilirubin, albumin, and insulin.

Results

It showed a significant decrease in serum levels of triglycerides (TGs), total cholesterol (T_C), low density lipoprotein cholesterol (LDL_C), liver alanine-, aspartate- and gamma glutamyl transaminase (ALT, AST, and GGT), total and direct bilirubin, fasting and post-prandial blood glucose, insulin, body weight and BMI while a significant increase in high density lipoprotein cholesterol (HDL_C) and albumin but non-significant change of alkaline phosphatase (ALP) in PO seeds treated subjects. Metformin (M) group has the same results of PO group except in high density lipoprotein cholesterol (HDL_C), LDL_C, and ALP levels had a different pattern.

Conclusions

PO seeds could be effective and safe as adjuvant therapy for Type-2 diabetic subjects. These results demonstrated that PO seeds possessed notable hypoglycaemic, hypolipidaemic and insulin resistance reducer effects; possibly due to its contents of polyunsaturated fatty acids, flavonoids, and polysaccharides.     

Prokinetic and laxative activities of Lepidium sativum seed extract with species and tissue selective gut stimulatory actions

Aim of the study

To provide ethnopharmacological basis for the medicinal use of Lepidium sativum seeds in indigestion and constipation.

Materials and methods

The in vivo studies were conducted in mice, while isolated tissues of mouse, guinea-pig and rabbit were suspended in tissue bath to measure isotonic contractions.

Results

The aqueous-methanolic extract of Lepidium sativum seeds (Ls.Cr) at 30 and 100 mg/kg showed atropine-sensitive prokinetic and laxative activities in mice, which were partially sensitive to atropine. In isolated gut preparations of mouse and guinea-pig, Ls.Cr (0.1-1 mg/mL) caused a concentration-dependent stimulatory effects both in jejunum and ileum, which was blocked in the presence of atropine. In rabbit jejunum, the stimulant effect of Ls.Cr remained unchanged in the presence of atropine, pyrilamine or SB203186, while in rabbit ileum, the stimulatory effect was partially blocked by atropine. The Ls.Cr was more efficacious in gut preparations of rabbit than in guinea-pig or mouse. The phytochemical analysis of the plant extract detected alkaloids, saponins and anthraquinones as plant constituents.

Conclusion

This study showed the prokinetic and laxative effects of Lepidium sativum in mice, which were partially mediated through a cholinergic pathway. The in vitro spasmodic effect of the plant extract mediated through a similar mechanism with species and tissue-selectivity, provides a rationale for the medicinal use of the seeds of Lepidium sativum in indigestion and constipation, and suggests studying the plant extracts on more than one species to get the wider picture.     

Protection against 1-methyl-4-phenylpyridinium ion (MPP+)-induced apoptosis by water extract of ginseng (Panax ginseng C.A. Meyer) in SH-SY5Y cells

Ethnopharmacological relevance

The present study investigates the protective effects of water extract of ginseng (Panax ginseng C.A. Meyer) against 1-methyl-4-phenylpyridinium ion (MPP⁺)-induced cytotoxicity in SH-SY5Y human neuroblastoma cells and explores the underlying mechanisms. The approach may be used for screening therapeutic agents for degenerative disorders such as Parkinson's disease.

Materials and methods

SH-SY5Y human neuroblastoma cells were used to analyze the protective effects of water extract of ginseng (WEG) against multiple parameters such as MPP⁺-induced viability, oxidative injury, expression of Bax, Bcl-2, cytochrome c and cleaved caspase-3.

Results

WEG exerted inhibitory effect on cell death, overproduction of ROS, elevated Bax/Bcl-2 ratio, release of cytochrome c and activation of caspase-3 expression in MPP⁺-treated SH-SY5Y cells.

Conclusions

WEG exhibited significant protective effects against MPP⁺-induced cytotoxicity in SH-SY5Y cells possibly through the suppression of ROS generation and the inhibition of mitochondria-dependent apoptotic pathway.     

Ethnobotanical study on some Malaysian anti-malarial plants: A community based survey

Ethnopharmacological relevance

Various plants species are used in the traditional medicine for the treatment of malaria. This is the first community based ethnobotanical study in Peninsular Malaysia.

Aim of the study

To investigate the plants traditionally used in the treatment of malaria in Malaysia.

Materials and methods

An ethnobotanical survey was carried out among 233 Aboriginal and rural households, and traditional healers in malaria endemic areas in Peninsular Malaysia. Data were collected using a pre-tested questionnaire.

Results

Nineteen species belonging to 17 families were identified. Twelve plant species have not previously been documented for the treatment of malaria in Malaysia.

Conclusions

Findings of this study can be used as an ethnopharmacological basis for selecting plants for further anti-malarial phytochemical and pharmaceutical studies.     

Sedative, hypnotic and anticonvulsant activities of the ethanol fraction from Rhizoma Pinelliae Praeparatum

Ethnopharmacological relevance

Rhizoma Pinelliae Praeparatum is the product of raw Rhizoma Pinellia processed with alkaline solution and Licorice, which had been widely used for treatment of insomnia in traditional Chinese medicine. The present study aimed to investigate the sedative, hypnotic and anticonvulsant activities of ethanol fraction from Rhizoma Pinelliae Praeparatum (EFRP) and to determine whether these effects were related to GABAergic mechanism.

Materials and methods

The sedative, hypnotic and anticonvulsant activities of EFRP were investigated with locomotion activity, pentobarbital-induced sleeping and nikethamide (NKTM)-induced convulsion tests, respectively. Additionally, the effects of flumazenil (an antagonist of GABA_A receptor) and l-malic acid (blocker of synthetic enzyme for GABA) on the hypnotic activity of EFRP were evaluated.

Results

EFRP at dose of 12 g/kg significantly inhibited the locomotion activity of mice. EFRP showed synergic effect on pentobarbital-induced sleeping by increased numbers of mice falling asleep, reduced the sleep latency and prolonged the sleeping time. l-malic acid and flumazenil inhibited the augment effects of EFRP on pentobarbital-induced sleeping. EFRP promoted a significant protection to NKTM-induced convulsion, by prolonged the death latency and decreased mortality.

Conclusion

EFRP possessed sedative, hypnotic and anticonvulsant activities and these activities may be related to the GABAergic system.     

The mechanism of vasorelaxation induced by ethanol extract of Sophora flavescens in rat aorta

Aim of the study

Sophora flavescens (SF) is a known medicinal herb for the treatment of cardiovascular symptoms associated with arrhythmia in China. However, the pharmacological action mechanisms involved have not been well studied. The aim of the present study was to define effects of roots of SF on the vascular tension and responsible mechanisms in rat thoracic aorta.

Materials and methods

Ethanol extract of the roots of SF (ESF) was examined for their vascular relaxant effect in isolated phenylephrine-precontracted rat thoracic aorta.

Results

ESF (0.1-100 μg/ml) induced relaxation of the phenylephrine-precontracted aortic rings in a concentration-dependent manner. Endothelium-denudation abolished the ESF-induced vasorelaxation. Pretreatment of the endothelium-intact aortic rings with l-NAME, an inhibitor of nitric oxide synthase, and ODQ, an inhibitor of soluble guanylyl cyclase (sGC), inhibited ESF-induced vasorelaxation. ESF increased cGMP levels of the aortic rings in a concentration-dependent manner and the effect was blocked by l-NAME and ODQ. Inhibition of K⁺ channels with glibenclamide and tetraethylammonium, cyclooxygenase inhibition with indomethacin, and β-adrenergic and muscarinic receptors blockade had no effect on the ESF-induced vasorelaxation.

Conclusion

These findings suggest that ESF relaxes vascular smooth muscle via endothelium-dependent NO-sGC-cGMP signaling pathway.     

Toxicity of Hydnora johannis Becca. dried roots and ethanol extract in rats

Aim of the study

Hydnora johannis Becca. (Hydnoraceae) commonly is used for the treatment of dysentery, diarrhoea, cholera and swelling tonsillitis in the folk medicine of Sudan and other African countries. This study evaluates the toxicological effects of Hydnora johannis roots on Wistar rats.

Materials and methods

Rats were randomized into control, groups fed with 2, 10, 20% of dried roots for 8 weeks and other groups given ethanol extract (50, 100, 200 and 400 mg/kg/day) through oral and intramuscularly administration for 2 weeks. Toxicity was evaluated using biochemical and histopathological assays.

Results

Alterations in the levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, cholesterol and urea were observed. Histopathological analysis revealed that the toxic effect were mainly on the liver, kidney and spleen on all treated groups. However, the impact of the dried roots was mild compared to the ethanol extract. Remarkably, there was a drop in cholesterol level in all treatment groups suggesting the antiartherogenic effect of Hydnora johannis roots.

Conclusion

The results from this study suggest that the powder preparation as well as ethanolic extract of Hydnora johannis roots induced toxic effect on Wistar rats. The observed toxic effect might be due to the dose and/or frequency of administration. Although in traditional medicine the extract is administrated in low dose, the results suggest the necessity of standardization of the drug.     

Evaluation of the anti-neuraminidase activity of the traditional Chinese medicines and determination of the anti-influenza A virus effects of the neuraminidase inhibitory TCMs in vitro and in vivo

Etnopharmacological relevance

Neuraminidase (NA) inhibitors are currently the most effective drugs to treat influenza A viruses infection. Many traditional Chinese medicines (TCMs) have been used in the clinics to treat influenza. The anti-viral mechanisms of these TCMs and their inhibitory effects towards NA need to be systematically tested.

Aim of the study

To evaluate the anti-NA activity of the TCMs and the anti-influenza A virus effects of the NA inhibitory TCMs in vitro and in vivo.

Material and methods

We tested the inhibitory activity of water extracts from 439 TCMs towards NA. The in vitro anti-influenza virus activities of the 5 TCMs were evaluated using the strain A/California/7/2009 (H1N1) NYMC X-179A of influenza A virus. A randomly selected TCM with NA inhibitory activity, Melia toosendan extract, was further evaluated using a mouse model infected with influenza A virus.

Results

Five TCMs, Duchesnea indica (Andr.) Focke [Fragaria indica Andr.], Liquidambar formosana Hance., Lithospermum erythrorhizon Sieb. et Zucc., Melia toosendan Sieb. et Zucc., and Prunella vulgaris L., exerted potent inhibitory activity towards NA. These TCMs in the range of 25-250 μg/mL had the ability to reduce virus-induced cytopathic effect (CPE) and the virus yield in MDCK cells. Melia toosendan significantly reduced death rate and prolonged mean day to death (MDD) of the viral infected mice.

Conclusions

This study describes five TCMs exerted strong inhibitory activities towards NA, and exhibited antiviral effect against influenza A virus by reducing viral reproduction and reduced CPE of the viral infected cells. Melia toosendan, significantly reduced death rate and prolonged survival of the H1N1 viral infected mice.     

Toxicity of medicinal plants used in traditional medicine in Northern Peru

Ethnopharmacological relevance

The plant species reported here are traditionally used in Northern Peru for a wide range of illnesses. Most remedies are prepared as ethanol or aqueous extracts and then ingested. The aim of this study was to evaluate the potential toxicity of these extracts.

Materials and methods

The toxicity of ethanolic and water extracts of 341 plant species was determined using a brine-shrimp assay.

Results

Overall 24% of the species in water extract and 76% of the species in alcoholic extract showed elevated toxicity levels to brine-shrimp. Although in most cases multiple extracts of the same species showed very similar toxicity values, in some cases the toxicity of different extracts of the same species varied from non-toxic to highly toxic.

Conclusions

Traditional preparation methods take different toxicity levels in aqueous and ethanol extracts into account when choosing the appropriate solvent for the preparation of a remedy.