Hepatoprotective activity of the edible snail (Bellamia bengalensis) flesh extract in carbon tetrachloride induced hepatotoxicity in rats

Ethnopharmacological relevance

In the folk-traditional medicine, snails were used to purify blood, boost immune system, prevent conjunctivitis and to treat liver problems.

Objectives

To evaluate the hepatoprotective activity of the edible snail (Bellamia bengalensis) flesh extract in male Wistar rats treated with carbon tetrachloride as an hepatotoxicant.

Materials and methods

Live adult Bellamia bengalensis was collected commercially from the Kolkata market. Aqueous flesh extract (BBE) was prepared in 0.9% saline and expressed in terms of wet weight basis. The aqueous flesh extract was administered orally (1, 2 g kg⁻¹ day⁻¹) to male rats for 12 days. Liv52 was used as positive control. 24 h after administration of extract, the rats were given a single oral dose of CCl₄ (1.25 ml kg⁻¹), except vehicle control rats. After 24 h of CCl₄ administration, all the animals were sacrificed to collect the blood and liver tissue.

Results

BBE (1 and 2 g kg⁻¹ day⁻¹, p.o. × 12 days) significantly prevented CCl₄ induced elevation of SGOT, SGPT, γGT, ACP, ALP, bilirubin, LDH and CCl₄ induced decrease in total protein, triglyceride level in male Wistar rats. BBE treated rat liver anti-oxidant parameters (LPO, SOD, GSH, CAT, GPx) were significantly antagonized for the pro-oxidant effect of CCl₄. Histopathological studies also supported the protective effect of BBE.

Conclusion

This study validated the folk and traditional use of snail in liver disorder through CCl₄-induced rat experimental model.     

Vasoactive effects of different fractions from two Panamanians plants used in Amerindian traditional medicine

Ethnopharmacological relevance

Cecropia obtusifolia (Cecropiaceae) and Psychotria poeppigiana (Synonym: Cephaelis elata, Rubiaceae) are two Latin American plants broadly used in traditional Amerindian medicine. The former, together with many other species of the genus Cecropia, share the folk reputation of curing heart failure, cough, asthma and bronchitis. The latter is used in Panama by Kuna and Ngäbe Buglé (Guaymies) native Indians for the treatment of dyspnea.

Aim of the study

Based on screening of selected medicinal Panamanian plants by radioligand-binding techniques by Caballero-George et al. (2001), the present study was carried out in order to investigate the vasoactive effects of different fractions from both P. poeppigiana and C. obtusifolia on rat thoracic aorta and identify active fractions and their chemical constituents.

Materials and methods

Both acid and neutral methanol fractions (P-AMeOH and P-NMeOH) and acid and neutral dichlorometane fractions (P-ADCM and P-NDCM) were obtained from P. poeppigiana crude methanolic and dichlorometane extracts, respectively. Identical fractionation was carried out for C. obtusifolia (C-AMeOH, C-NMeOH, C-ADCM and C-NDCM. Vasorelaxant effect of all fractions, and their inhibition of contractile responses to angiotensin II were evaluated in isolated aortic rings.

Results

P-AMeOH, P-NMeOH and P-ADCM fractions induced a concentration-dependent relaxation (43.9 ± 1.8%, 35.3 ± 4.7% and 52.9 ± 3.5%, respectively) in the endothelium-intact aorta precontracted by phenylephrine (PE, 10⁻⁶ M). The relaxation produced by C-AMeOH and C-NMeOH (57.3 ± 2.5% and 53.3 ± 3.3%, respectively) was greater than the effect produced by C-ADCM and C-NDCM (42.2 ± 3.4% and 21.8 ± 0.8%, respectively). Only the incubation of the aortic rings with P-AMeOH reduced the maximum contraction induced by angiotensin II at 20.08 ± 0.55%.

Conclusions

The direct vasorelaxation effect observed could explain in part the ethnomedical use of these plants in Amerindian traditional medicine. The most active fractions contain phenolic and aromatic acid compounds. Furthermore, P-AMeOH, the only fraction that showed both vasorelaxant effect and inhibition of contractile responses to angiotensin II, is the most rich in aromatic acids compounds and the only one that contains scopoletin.     

Oral Uncaria rhynchophylla (UR) reduces kainic acid-induced epileptic seizures and neuronal death accompanied by attenuating glial cell proliferation and S100B proteins in rats

Aim of the study

Epilepsy is a common clinical syndrome with recurrent neuronal discharges in cerebral cortex and hippocampus. Here we aim to determine the protective role of Uncaria rhynchophylla (UR), an herbal drug belong to Traditional Chinese Medicine (TCM), on epileptic rats.

Materials and methods

To address this issue, we tested the effect of UR on kainic acid (KA)-induced epileptic seizures and further investigate the underlying mechanisms.

Results

Oral UR successfully decreased neuronal death and discharges in hippocampal CA1 pyramidal neurons. The population spikes (PSs) were decreased from 4.1 ± 0.4 mV to 2.1 ± 0.3 mV in KA-induced epileptic seizures and UR-treated groups, respectively. Oral UR protected animals from neuronal death induced by KA treatment (from 34 ± 4.6 to 191.7 ± 48.6 neurons/field) through attenuating glial cell proliferation and S100B protein expression but not GABAA and TRPV1 receptors.

Conclusions

The above results provide detail mechanisms underlying the neuroprotective action of UR on KA-induced epileptic seizure in hippocampal CA1 neurons.     

Screening of Caesalpinia bonduc leaves for antipsoriatic activity

Ethnopharmacological relevance

Leaves of Caesalpinia bonduc (L.) Roxb. (Caesalpiniaceae) have been used by traditional Siddha healer of Malabar region for psoriasis treatment.

Aim of the study

To evaluate the Caesalpinia bonduc decoction (CBD), Caesalpinia bonduc hydroalcoholic extract (CBHA) for antipsoriatic activity.

Materials and methods

Mouse tail test for psoriasis was used for the evaluation of antipsoriatic activity. Extracts were tested at a dose of 500 mg/kg b.w. and fractions at 250 mg/kg b.w. in Swiss albino mice. Parameters studied in the mouse tail test were changes in epidermal thickness and percentage orthokeratotic values. In vitro antiproliferant assay on HaCaT cell lines and in vitro lipoxygenase inhibition were also carried out.

Results

Butanol fraction of Caesalpinia bonduc hydroalcoholic extract (CBHAB) and water fraction of Caesalpinia bonduc hydroalcoholic extract (CBHAW) produced significant orthokeratosis (p < 0.001). In relative epidermal thickness, a significant (p < 0.05) reduction with respect to control was observed in groups treated with retinoic acid, CBD, butanol fraction of Caesalpinia bonduc decoction (CBDB), water fraction of Caesalpinia bonduc hydroalcoholic extract (CBHAW). Maximum antiproliferant activity was shown by CBHA (IC₅₀, 77.5 ± 12.7 μg/ml). In lipoxygenase inhibition assay, water fraction of Caesalpinia bonduc decoction (CBDW) showed maximum activity with an IC₅₀ value of 164.71 ± 4.57 μg/ml.

Conclusions

Among all the tested samples only CBHAW showed good activity in the mouse tail test, antiproliferant activity in HaCaT cells and lipoxygenase inhibition assay. Other extracts and fractions showed varying degrees of activity. The present study supports the traditional use of Caesalpinia bonduc leaves for psoriasis treatment.     

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.     

The potential role of female flowers inflorescence of Typha domingensis Pers. in wound management

Aim of the study

Female flowers inflorescence of Typha species including Typha domingensis Pers. are used externally for burns and wound healing in Turkish folk medicine. In order to verify the folkloric assertion, the female and male flowers inflorescences were individually submitted to in vivo wound models.

Material and methods

Ointment formulations prepared directly either from the male or female flowers inflorescences of Typha domingensis in 5% and 10% concentrations were submitted to activity testing. After that, female flowers inflorescence was further submitted to successive extractions with solvents in increasing polarity; i.e., n-hexane, chloroform, methanol and water and the wound healing activity of each extract was investigated. The linear incision and circular excision wound models were used for the evaluation of the healing potential of the test materials in rats and mice. Tissue sections were also evaluated by histopathological techniques.

Results

Remarkable wound healing activity was observed only for the female flowers inflorescence at 5% concentration in ointment base and its methanolic and aqueous extracts. The wound healing effect was found comparable to that of reference ointment Madecassol^®. The results of histopathological evaluation supported the outcome of both linear incision and circular excision wound models.

Conclusion

The experimental study revealed that the female flowers inflorescence of Typha domingensis displayed notable wound healing activity in mice and rats, at the models tested.     

Cordia verbenacea and secretion of mast cells in different animal species

Ethnopharmacology relevance

Different plant species from Cordia genera are used in folk medicine as anti-inflammatory medication throughout the tropical and subtropical regions of the world. In Brazil, Cordia verbenacea is a medicinal plant known as “erva-baleeira”. The alcoholic extracts, decoctions and infusions with leaves of C. verbenacea are used in Brazilian traditional medicine for treatment of cough, pneumonia, parasitic diseases and, especially, the inflammatory processes. Anti-inflammatory activity was already demonstrated; however, molecular mechanisms of action are not completely understood. Considering the importance of histamine in early events of inflammation and in allergic diseases, we evaluated the effect of ethanol extract of leaves of C. verbenacea on histamine release (in vitro and in vivo studies) from different types of mast cells induced by chemical agents using several species of rodents.

Materials and Methods

The extraction and quantification of histamine were performed by using an automatic fluorometric continuous flow system.

Results

The extract of C. verbenacea (30 μg/ml) reduced the in vitro secretion of histamine from rat mast cells induced by ionophore A23187, concanavalin A and compound 48/80, respectively, to 22.1 ± 2.2%, 24.3 ± 2.5% and 21.4 ± 2.1%. At the same concentration, the extract also inhibited the secretion of histamine from mast cells of guinea pig induced by ionophore A23187 to 33.3 ± 2.2%, and mast cells of hamster induced by ionophore A23187 and concanavalin A to 15.8 ± 2.5% and 10.8 ± 2.6%, respectively. The oral treatment with the extract (300 mg/kg) also inhibited the secretion of histamine induced by A23187 about to 36.3 ± 3.2% in rats.

Conclusions

C. verbenacea inhibits the in vitro secretion of histamine from mast cells of different animal species, as well as the secretion of mast cells from animals treated with the extract, which gives not only the proven anti-inflammatory effect of the plant, but also anti-allergic effect, opening new possibilities for future anti-allergic herbal medicine.     

Healing potential of Rosmarinus officinalis L. on full-thickness excision cutaneous wounds in alloxan-induced-diabetic BALB/c mice

Aim of the study

Rosmarinus officinalis (Rosemary) used in Jordanian folk medicine for wound management and treatment. Therefore, the present study was conducted to assess the healing efficacy of both aqueous extract and essential oil of the aerial parts on alloxan-induced diabetic BALB/c mice.

Materials and methods

Two full-thickness round wounds were created in the dorsal area of each mouse. Animals were divided into four groups of twenty mice each: untreated normal, untreated diabetic, aqueous extract- (intraperitoneal injection of 0.2 ml at a dose of 10% for 3 days) and essential oil-treated diabetic mice (topical application of 25 μl/excision wound, twice a day for 3 days). For 15 days, the wounds were visually observed; blood glucose level, body weight, regenerated granulation tissue weight and the percentage of wound contraction were measured. On days 6 and 15 after wounding, the animals were sacrificed and the histology of wound area was examined.

Results

Significant positive differences (p < 0.01) between treated and control groups were observed at different aspects of diabetic wound healing process. Reduced inflammation and enhanced wound contraction, re-epithelialization, regeneration of granulation tissue, angiogenesis and collagen deposition were detected in the treated wounds.

Conclusions

Results indicated that the essential oil of Rosmarinus officinalis was the most active in healing diabetic wounds and provide a scientific evidence for the traditional use of this herb in wound treatment. However, further scientific verification is required to confirm and assess the range of wound healing potential of essential oils of Rosemary chemotypes.     

Anti-inflammatory activity of Terminalia paniculata bark extract against acute and chronic inflammation in rats

Ethnopharmacological relevance

Terminalia paniculata Roxb. (Family-Combretaceae) is a wild tree commonly used in traditional ayurvedic medicine for the treatment of inflammation of parotid glands and in menstrual disorders.

Aim of the study

To explore the folk use of Terminalia paniculata on pharmacological grounds to evaluate the scientific basis of anti-inflammatory activity.

Materials and methods

The anti-inflammatory activity of Terminalia paniculata was studied against carrageenan-induced hind paw edema, air pouch inflammation and complete Freund's adjuvant (CFA)-induced arthritis in rats. The aqueous extract of Terminalia paniculata bark (TPW) was administered at the concentrations of 100, 200 and 400 mg/kg body weight.

Results

TPW showed significant (p < 0.05) anti-inflammatory activity by reducing the edema volume in carrageenan-induced paw edema in rats. Further, TPW (400 mg/kg) also reduced the carrageenan-induced leukocyte migration (50.92 ± 5.71%) and myeloperoxidase activity (49.31 ± 5.24%) in air pouch exudates. TPW (200 mg/kg) exhibits anti-rheumatic and analgesic activities by improving the altered haematological milieu (ESR, CRP, RF, WBC, RBC and Hb) and also by inhibiting the flexion scores and radiographic changes in CFA-induced arthritis. This extract also had significant (p < 0.05) effects on the occurrence of secondary lesions compared to CFA control.

Conclusions

Terminalia paniculata bark may be a potential preventive or therapeutic candidate for the treatment of chronic inflammation and arthritis.     

Permeability of rosmarinic acid in Prunella vulgaris and ursolic acid in Salvia officinalis extracts across Caco-2 cell monolayers

Ethnopharmacological relevance

Rosmarinic acid (RA), a caffeic acid-related compound found in high concentrations in Prunella vulgaris (self-heal), and ursolic acid (UA), a pentacyclic triterpene acid concentrated in Salvia officinalis (sage), have been traditionally used to treat inflammation in the mouth, and may also be beneficial for gastrointestinal health in general.

Aim of the study

To investigate the permeabilities of RA and UA as pure compounds and in Prunella vulgaris and Salvia officinalis ethanol extracts across human intestinal epithelial Caco-2 cell monolayers.

Materials and methods

The permeabilities and phase II biotransformation of RA and UA as pure compounds and in herbal extracts were compared using Caco-2 cells with HPLC detection.

Results

The apparent permeability coefficient (P_app) for RA and RA in Prunella vulgaris extracts was 0.2 ± 0.05 × 10⁻⁶ cm/s, significantly increased to 0.9 ± 0.2 × 10⁻⁶ cm/s after β-glucuronidase/sulfatase treatment. P_app for UA and UA in Salvia officinalis extract was 2.7 ± 0.3 × 10⁻⁶ cm/s and 2.3 ± 0.5 × 10⁻⁶ cm/s before and after β-glucuronidase/sulfatase treatment, respectively. Neither compound was affected in permeability by the herbal extract matrix.

Conclusion

RA and UA in herbal extracts had similar uptake as that found using the pure compounds, which may simplify the prediction of compound efficacy, but the apparent lack of intestinal glucuronidation/sulfation of UA is likely to further enhance the bioavailability of that compound compared with RA.     

Antidepressant effect of Valeriana wallichii patchouli alcohol chemotype in mice: Behavioural and biochemical evidence

Ethnopharmacological relevance

Valeriana wallichii DC, an ayurvedic traditional medicine has now been shown to exist chemically as three distinct chemotypes. The study aimed to investigate the antidepressant effect of dichloromethane extract of Valeriana wallichii patchouli alcohol chemotype.

Materials and methods

Antidepressant effect of dichloromethane extract of Valeriana wallichii (10, 20 and 40 mg/kg, p.o.) using forced swim test, was determined in both acute and chronic study. The neurotransmitter levels were estimated in mouse forebrain after two weeks of dosing.

Results

Single administration of extract (40 mg/kg) significantly inhibited the immobility period in mice (p < 0.05). Similarly, chronic administration of extract (20 and 40 mg/kg) significantly reduced the immobility period and significantly increased the levels of norepinephrine and dopamine in mouse forebrain (p < 0.05).

Conclusions

The extract demonstrated antidepressant effect and significantly increased the norepinephrine and dopamine levels in forebrain.     

Cytochrome P450 inhibitory potential of Triphala--a Rasayana from Ayurveda

Ethnopharmacological relevance

‘Triphala’ is one of the age-old, most commonly used polyherbal preparation from Ayurveda as Rasayana drug.

Aim of the study

This study was aimed at evaluating the effect of ‘Triphala’ on drug modulating enzymes to assess its safety through its potential to interact with co-administered drugs.

Materials and methods

The cytochrome P450 inhibitory effect of ‘triphala’ formulation was investigated on rat liver microsomes using CYP450-CO complex assay and on individual isoform such as CYP3A4 and 2D6 using fluorescence screening. RP-HPLC method was developed to standardize ‘triphala’ and its individual components using gallic acid as analytical marker compound.

Results

RP-HPLC analysis demonstrated the presence of gallic acid (4.30 ± 2.09 mg/g) in the formulation. The formulation showed 23% inhibition of the rat liver microsomes through CYP450-CO complex assay which is comparatively less when compared with the individual components. Further, the effect of standardized formulation dissolved in ethanol showed CYP3A4 and CYP2D6 inhibitory activity at the IC₅₀ values of 119.65 ± 1.91 μg/ml and 105.03 ± 0.98 μg/ml respectively. Gallic acid was also found to inhibit both the isoforms at the IC₅₀ values of 87.24 ± 1.11 μg/ml and 92.03 ± 0.38 μg/ml respectively.

Conclusions

Various concentrations of the formulation and its individual components showed significantly less inhibitory activity (p < 0.001) on individual isoforms when compared with the positive control. Assessment on the in vitro effect of ‘triphala’ on drug modulating enzymes has important implications for predicting the likelihood of herb-drug interactions if these are administered concomitantly.     

Evaluation of the acute dermal exposure of the ethanolic and hexanic extracts from leaves of Schinus molle var. areira L. in rats

Ethnopharmacological relevance

Schinus molle var. areira L. (Anacardiaceae) is employed in herbal medicine for many conditions, including respiratory, urinary and menstrual disorders, and as a digestive stimulant, diuretic, astringent and antidepressant. It is also known for its topical use as wound healer, antiseptic, for skin disorders and as repellent and insecticide. In the present work, the acute dermal exposure to ethanolic and hexanic extracts from leaves of Schinus molle var. areira was studied in rats.

Materials and methods

A single dose of 2000 mg/kg of body weight of ethanolic and hexanic extracts from leaves was applied on the shaved skin of male and female rats. After 24 h of exposure, the patch was removed and any sign of irritation was recorded. Behavioral and functional parameters in a functional observational battery and motor activity in an open field were assessed after the exposure to the extracts. Then, after 14 days of observation, animals were retested. Finally, histopathological studies were conducted on several organs.

Results

Slight signs of erythema and edema were observed in the skin site of exposure, but they disappeared after 48 h. The exposure to the hexanic extract produced an increase in parameters of activity, rearing and arousal assessed in the functional observational battery, which reversed after 14 days. On the other hand, the ethanolic extract caused an increase in locomotor activity, reflected in a higher number of rearings performed in the open field in the evaluation carried out on Day 14. No histopathological alterations were detected in the analyzed organs.

Conclusions

The results show that the acute dermal exposure of the ethanolic and hexanic extracts from leaves of Schinus molle var. areira only causes a slight and reversible skin irritation, and a mild stimulatory effect in rats. All these indicate that the topical use of these extracts would be safe.     

Antidiabetic potential of unripe Carissa carandas Linn. fruit extract

Ethnopharmacological relevance

Carissa carandas commonly known as Karanda have a long history of use in traditional system of medicine. It is used by tribal healers of Western Ghat region of Karnataka as hepatoprotective and antihyperglycemic. However, no scientific data is available to validate the folklore claim. The present study has been designed to evaluate its unripe fruit for the antidiabetic activity.

Aim

In the present study, methanol extract of unripe fruits and its fractions were studied for its antidiabetic potential.

Materials and methods

The methanol extract and its fractions were screened for antidiabetic activity in alloxan induced diabetic rats. The polyphenolic, flavonoid and flavanone contents of methanolic extract and its fractions were also determined and correlated with its antidiabetic activity.

Results

The experimental data indicated that the methanol extract and its ethyl acetate soluble fraction has significantly lowered the elevated blood glucose levels by 48% (p < 0.001) and 64.5% (p < 0.001) respectively at dose level of 400 mg/kg per oral after 24 h as compared to diabetic control. In order to assess the role of polyphenolic components in the relevant activity, polyphenolic and flavonoid contents were determined. The polyphenolic and flavonoid content of methanol extract and its ethyl acetate soluble fraction were found to be 15.8 ± 1.2 mg and 18.55 ± 0.34 mg (gallic acid equivalent/g extract) and flavonoid content 2.92 ± 0.03 mg and 1.534 ± 0.30 mg (rutin equivalent/g extract) respectively.

Conclusion

The increased antidiabetic potential of ethyl acetate fraction over methanol extract is due to its partial purification achieved by fractionation which resulted in increase in degree of polymerization and segregation of secondary metabolites.     

5α-reductase inhibition and hair growth promotion of some Thai plants traditionally used for hair treatment

Ethnopharmacological relevance

Many Thai traditional herbs have been used for hundreds of years for hair treatment and nourishment, including hair loss. However, scientific evidence about their mechanisms of action has not yet been elucidated.

Aims of the study

The purpose of this research is to define the possible mechanisms involved in hair loss treatment of the selected plants by determining the 5α-reductase enzyme inhibition and hair growth promoting activities, and the relationship between these two activities.

Materials and methods

Seventeen Thai plants traditionally used for hair treatment were selected. The plants were dried, ground and extracted by maceration with ethyl alcohol. These extracts were further tested for 5α-reductase inhibition using enzymes from rat livers. Hair growth promoting activity was tested in C57BL/6 mice.

Results

Carthamus tinctorius L. was the most potent 5α-reductase inhibitor, with a finasteride equivalent 5α-reductase inhibitory activity (FEA) value of 24.30 ± 1.64 mg finasteride equivalent per 1 g crude extract. Phyllanthus emblica L. was the second most potent inhibitor, with FEA of 18.99 ± 0.40. Rhinacanthus nasutus (L.) Kurz. was the least potent 5α-reductase inhibitor (FEA 10.69 ± 0.96). Carthamus tinctorius also was the most potent hair growth promoter in C57BL/6 mice. There were strong relationships between 5α-reductase inhibitory activity and hair growth promoting activity (r = 0.719), and between 5α-reductase inhibitory activity and hair follicle count (r = 0.766).

Conclusions

Ethanolic extract of Carthamus tinctorius was the most potent 5α-reductase inhibitor and hair growth promoter. This discovery may lead to the development of new alternative medicines for hair loss prevention and treatment.     

Bronchospasm potentiating effect of methanolic extract of Ficus religiosa fruits in guinea pigs

Ethnopharmacological relevance

The sacred tree Peepal (Ficus religiosa family: Moraceae) has numerous therapeutic utility in folk medicine.

Aim of the study

It has been reported to be used in ethno medical treatment of asthma and also in epilepsy due to its high serotonin content, which has been implicated in pathophysiology of asthma, this led us to carry out the present study.

Materials and methods

The in vivo studies of histamine induced bronchospasm in guinea pigs and in vitro isolated guinea pig tracheal chain and ileum preparation.

Results

Pre-treatment of guinea pigs with ketotifen (1 mg/kg, p.o.) has significantly delayed the onset of histamine aerosol induced pre-convulsive dyspnea, compared with vehicle control (281.8^a ± 11.7 vs. 112.2 ± 9.8). The administration of methanolic extract (125, 250 and 500 mg/kg, p.o.) did not produced any significant effect on latency to develop histamine induced pre-convulsive dyspnea. On the other hand, methanolic extract of the fruits at the doses employed (i.e., 0.5, 1 and 2 mg/ml) has significantly potentiate the EC₅₀ doses of both histamine and acetylcholine in isolated guinea pig tracheal chain and ileum preparation. In addition, HPLC analysis of the methanolic extract showed the presence of high amounts of serotonin (2.89%, w/w).

Conclusions

On the basis of data, it may be concluded that Ficus religiosa fruits have been found to be ineffective against histamine induced bronchospasm in guinea pigs. In addition, methanolic extract of the fruits have shown to potentiate the bronchoconstriction induced by both histamine and acetylcholine on guinea pig tracheal chain preparation.     

LC-MS/MS determination and pharmacokinetic study of five flavone components after solvent extraction/acid hydrolysis in rat plasma after oral administration of Verbena officinalis L. extract

Ethnopharmacological relevance

Traditional Chinese medicine (TCM) has been used in clinical practice for several thousand years. TCM has played an indispensable role in the prevention and treatment of diseases, especially the complicated and chronic ones. Pharmacokinetic study on active constituents in herbal preparations is a good way for us to explain and predict a variety of events related to the efficacy and toxicity of TCM.

Aim of the study

A selective and sensitive HPLC-MS/MS method was first developed and validated for the determination of luteolin, kaempferol, apigenin, quercetol, and isorhamnetin in rat plasma.

Materials and methods

The LC system consisted of an Agilent Technologies Series 1200 system (Agilent, USA) equipped with an automatic degasser, a quaternary pump, and an autosampler. Chromatographic separations were performed on a Waters SunFire™ C₁₈ column (150 mm × 4.6 mm, 5 μm), and the column temperature was maintained at 25 °C and the sample injection volume was 20 μL. The current LC-MS/MS assay was validated for linearity, intra-day and inter-day precisions, accuracy, extraction recovery and stability.

Results

The validated method was successfully applied to monitoring the concentrations and pharmacokinetic studies of five flavone compounds in rat plasma after a single oral administration of Verbena officinalis L. extract with a dosage of 8.0 mL/kg. The time to reach the maximum plasma concentration (T_max1) was 0.48 ± 2.14 h for luteolin, 0.25 ± 0.13 h for kaempferol, 0.97 ± 1.08 h for apigenin, 1.04 ± 4.25 h for quercetol and 0.25 ± 0.16 h for isorhamnetin, and the maximum plasma concentration (T_max2) was 3.97 ± 1.48 h, 4.05 ± 0.46 h, 4.33 ± 0.58 h, 2.99 ± 0.48 h and 4.02 ± 0.34 h. The elimination half-time (t_1/2) of luteolin, kaempferol, apigenin, quercetol and isorhamnetin was 4.02 ± 0.81, 7.65 ± 0.71, 3.30 ± 0.83, 4.55 ± 0.49 and 5.56 ± 1.32 h, respectively.

Conclusions

This paper described a simple, sensitive and validated LC-MS/MS method for simultaneous determination of luteolin, kaempferol, apigenin, quercetol and isorhamnetin in rat plasma after oral administration of V. officinalis L. extract, and investigated on their pharmacokinetic studies as well, with a short run time of 5 min.     

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.     

Sida rhomboidea.Roxb leaf extract ameliorates gentamicin induced nephrotoxicity and renal dysfunction in rats

Ethnopharmacological relevance

Sida rhomboidea.Roxb (SR) known as “Mahabala” in Ayurveda and marketed as “Shahadeyi” is used in ethnomedicine to treat ailments such as dysuria and urinary disorders.

Aim of the study

To evaluate nephroprotective potential of SR against gentamicin (GM) induced nephrotoxicity and renal dysfunction.

Materials and methods

Nephrotoxicity was induced in rats with GM (100 mg/kg bodyweight (i.p.) for 8 days) and were treated with SR extract (200 and 400 mg/kg bodyweight (p.o.) for 8 days) or 0.5% carboxymethyl cellulose (vehicle). Plasma and urine urea and creatinine, renal enzymatic and non-enzymatic antioxidants along with lipid peroxidation were evaluated in various experimental groups.

Results

GM treatment induced significant elevation (p < 0.05) in plasma and urine urea, creatinine, renal lipid peroxidation along with significant decrement (p < 0.05) in renal enzymatic and non-enzymatic antioxidants. SR treatment to GM treated rats (GM + SR) recorded significant decrement (p < 0.05) in plasma and urine urea and creatinine, renal lipid peroxidation along with significant increment (p < 0.05) in renal enzymatic and non-enzymatic antioxidants.

Conclusion

SR leaf extract ameliorates GM induced nephrotoxicity and renal dysfunction and thus validates its ethnomedicinal use.     

Pharmacological characterization of the diuretic effect of Hibiscus sabdariffa Linn (Malvaceae) extract

Ethnopharmacological relevance

Hibiscus sabdariffa L. (Malvaceae) populary known in Mexico as “Jamaica”, “flor de Jamaica”, has widely used in Mexican Traditional Medicine as antihypertensive and diuretic, although the latter activity has been reported the present work show evidence about the diuretic, natriuretic and potassium-sparing effects.

Aim of the study

To evaluate the diuretic activity of Hibiscus sabdariffa aqueous extract on in vivo and in situ models.

Materials and methods

The Hibiscus sabdariffa aqueous extract was administrated in increasing doses and evaluated the diuresis produced and disposal of electrolytes. Moreover, in isolated kidney was determined the renal filtration rate with plant extract, furosemide and amiloride.

Results

The yield of Hibiscus sabdariffa aqueous extraction was 28.3% and the chemical standardization from 1 g of extract was: 56.5 mg delphinidin-3-O-sambubioside, 20.8 mg/g cyanidin-3-O-sambubioside, 3.2 mg/g quercetin, 2.1 mg/g rutin and 2.7 mg/g chlorogenic acid. The diuretic and natriuretic effect of Hibiscus sabdariffa aqueous extract showed a dose-dependent behavior. The pharmacological constants of natriuretic effect was ED50 = 86 mg/kg and Emax = 0.9 mEq/100 g/5 h. In the model of kidney in situ was observed that renal filtration increased 48% with the aqueous extract of Hibiscus sabdariffa and an additive effect when was perfuse with furosemide.

Conclusion

The compound presents in Hibiscus sabdariffa as quercetin had effect on the vascular endothelium causing oxide nitric release, increasing renal vasorelaxation by increasing kidney filtration. Therefore, the diuretic effect of Hibiscus sabdariffa may be mediated by nitric oxide release.