In vitro modulatory effects of Andrographis paniculata, Centella asiatica and Orthosiphon stamineus on cytochrome P450 2C19 (CYP2C19)

Ethno pharmacological relevance

Andrographis paniculata (AP), Centella asiatica (CA) and Orthosiphon stamineus (OS) are three popular herbs traditionally used worldwide. AP is known for the treatment of infections and diabetes and CA is good for wound healing and healthy skin while OS is usually consumed as tea to treat kidney and urinary disorders. Interaction of these herbs with human cytochrome P450 2C19 (CYP2C19), a major hepatic CYP isoform involved in metabolism of many clinical drugs has not been investigated to date.

Aim of the study

In this study, the modulatory effects of various extracts and major active constituents of AP, CA and OS on CYP2C19 activities were evaluated.

Materials and methods

S-Mephenytoin, the CYP2C19 substrate probe, was incubated in the presence or absence of AP, CA and OS components. The changes in the rate of metabolite (hydroxymephenytoin) formation were subsequently determined by a high-performance liquid chromatography (HPLC)-based enzyme assay to characterize the modulatory effects.

Results

Among the herbal extracts studied, AP ethanol extract and CA dichloromethane extract exhibited mixed type inhibition towards CYP2C19 with K_i values of 67.1 and 16.4 μg/ml respectively; CA ethanol extract and OS petroleum ether extract competitively inhibited CYP2C19 activity (K_i = 39.6 and 41.5 μg/ml respectively). Eupatorin (a major active constituent of OS) was found to significantly inhibit CYP2C19 by mixed type inhibition (K_i = 7.1 μg/ml or 20.6 μM).

Conclusions

It was observed that AP, CA and OS inhibited CYP2C19 activity with varying potency. While weak inhibitory effect was observed with AP, moderate to strong inhibition was observed with CA dichloromethane extract and eupatorin, the major OS constituent. Therefore care should be taken when these CA and OS components are co-administered with CYP2C19 substrates (such as omeprazole, proguanil, barbiturates, citalopram, and diazepam).     

Dual protective effect of Passiflora incarnata in epilepsy and associated post-ictal depression

Ethnopharmacological relevance

Passiflora incarnata L. (Passifloraceae) has been used for the treatment of epilepsy in several traditional systems of medicine.

Aim of the study

The aerial parts of Passiflora incarnata contain multiple bioactive metabolites such as, flavonoids (like, chrysin that show CNS depressant activity by agonizing GABA-benzodiazepine receptor), amino acids (like, GABA), harmala alkaloids (reversible monoamine oxidase-A inhibitor), etc. In view of this, the present study was designed to investigate dual protective effect of the hydroethanolic extract of Passiflora incarnata in pentylenetetrazol (PTZ)-induced seizure and associated post-ictal depression.

Materials and methods

Different groups of mice were administered with repeated subconvulsive doses of PTZ (50 mg/kg; i.p.) at an interval of 5 days for 15 days. From 5th to 15th day the animals in different groups were administered daily with varying doses of hydroethanolic extract of Passiflora incarnata (150, 300, and 600 mg/kg; i.p.), diazepam (2 mg/kg; i.p.) and vehicle. On every 5th day, after PTZ treatment, seizure severity (score) was noted. Following convulsive episodes the locomotor activity (using actophotometer) and immobility period (using forced swim test) were also determined. On 15th day after behavioral assessment, the brain serotonin and noradrenaline levels were determined using spectrofluorometric methods.

Results

Treatment with the extract significantly (p < 0.05) reduced the seizure severity and immobility period as compared to vehicle control, in a dose and time-dependent manner. Moreover, the extract treatment retained the serotonin and noradrenaline levels of the brain.

Conclusions

The results of present study concluded that the hydroethanolic extract of Passiflora incarnata suppress PTZ-induced seizures, and ameliorates its associated post-ictal depression, which has been found to be get worsened with the standard antiepileptic drug, diazepam.     

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.     

Wound healing activity of Glycosmis arborea leaf extract in rats

Ethnopharmacological relevance

Glycosmis arborea Roxb, syn. Glycosmis pentaphylla Retz, (Rutacea) is a shrub found all over India, have been used for the healing of wounds of livestock in Indian folk medicine.

Aim of the study

The present study provides a scientific evaluation for the wound healing potential of ethanolic extract of Glycosmis arborea leaves.

Methods and materials

Excision and incision wounds were inflicted upon four groups containing six rats each. Group-1 was assigned as control (ointment base), Group-2 and Group-3 were treated with 5 and 10% EtOH extract ointment. Group-4 was treated with standard Nitrofurazone (0.2%) ointment. The parameters observed were percentage of wound contraction, epithelization period, tensile strength, hydroxyproline content. Ethanolic extract of Glycosmis arborea leaves was given orally at a dose of 200, 400 mg/kg/day in dead space wound model.

Results

Topical application of Glycosmis arborea ointment (10%) in excision wound model significantly (P < 0.05) increased the percentage of wound contraction (96.91%) compared with control (60.27%) and decreased the epithelization time. The tensile strength significantly (P < 0.05) increased in Group-3 at 560.33 ± 6.48 g when compared to control at 319.17 ± 6.16 g. Rats treated with Glycosmis arborea extract (400 mg/kg) showed significant (P < 0.05) increase in hydroxyproline content at 54.94 ± 0.96 mg/g when compared with control at 30.77 ± 1.13 mg/g.

Conclusion

The ethanolic extract of Glycosmis arborea facilitated wound healing significantly, corroborating the folk medicinal use of this plant.     

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.     

Inhibition of glucose-induced vascular endothelial growth factor expression by Salvia miltiorrhiza hydrophilic extract in human microvascular endothelial cells: evidence for mitochondrial oxidative stress

Aim of study

Diabetes mellitus is frequently combined with vascular diseases, which are associated with the expression of vascular endothelial growth factor (VEGF). An approach that can reverse the induction of VEGF by hyperglycemia may potentially benefit the outcome of diabetic patients. Therefore, in the present study, we investigated the effect of Salvia miltiorrhiza (S. miltiorrhiza) hydrophilic extract on the expression of VEGF induced by high concentration of glucose.

Materials and methods

Vector of VEGF promoter luc was transiently transfected into HMEC-1 cells, and luciferase activity was measured to determine the promoter activity. In order to investigate the mechanism of Salvia miltiorrhiza hydrophilic extract, mitochondrial uncoupling protein 2(UCP2) was knockdown by using UCP2 siRNA. The expression of VEGF was obtained by using quantitative RT-PCR and dot blot. The level of reactive oxygen species (ROS) was expressed by the level of 2′,7′-dichlorfluorescein.

Results

Exposure of HMEC-1 cells to 30 mM glucose resulted in a significant increase in the expression of VEGF mRNA (5.7 fold at 3 mM glucose, P < 0.005), and an increase of ROS formation (2.4 fold at 3 mM glucose, P < 0.005). These effects were completely antagonized by an inhibitor of electron transport chain complex II, thenoyltrifluoroacetone (TTFA) and an uncoupler of oxidative phosphorylation, carbonylcyanide-m-chlorophenylhydrazone (CCCP). Addition of Salvia miltiorrhiza hydrophilic extract (10 μg/ml) led to a significant decrease of VEGF mRNA and ROS formation in 30 mM glucose condition. Interestingly, knockdown of mitochondrial UCP-2 by UCP-2 siRNA abolished the reduction of VEGF expression and ROS formation by Salvia miltiorrhiza hydrophilic extract.

Conclusions

These findings indicated that Salvia miltiorrhiza hydrophilic extract effectively reversed induction of VEGF expression by high glucose via ameliorating mitochondrial oxidative stress. Salvia miltiorrhiza hydrophilic extract can potentially be an effective antioxidant therapy for the treatment of diabetic chronic vascular complication.     

Antidiabetic activity of alcoholic stem extract of Nervilia plicata in streptozotocin-nicotinamide induced type 2 diabetic rats

Ethnopharmacological relevance

Nervilia plicata (Orchidaceae) has long been used in the antidiabetic medicinal preparations of traditional healers of Wayanad (Kerala), but recuperative potential of the plant was remained undefined. We demonstrated the regenerative potential of the plant extract on kidney affected by type 2 diabetes besides lowering blood glucose.

Aim of the study

The aim of the current study was to investigate the recuperative and regenerative potential of alcoholic stem extract of Nervilia plicata on streptozotocin-nicotinamide induced type 2 diabetic models.

Materials and methods

Non insulin dependent diabetes mellitus (NIDDM) was induced in overnight fasted rats by intramuscular injection (IMI) of 60 mg/kg STZ and 120 mg/kg of nicotinamide after 5 min interval. Blood glucose was assessed by a glucometer, serum urea and creatinine levels were determined by diacetylmonooxime method and Jaffe reaction respectively. Kidney sections were taken and stained with Masson's tri-dye and Periodic Acid Schiff (PAS) and examined for structural changes. Also lipid peroxidation product (LPP) levels were determined as thio barbituric acid reactive substance levels (TBARS) method.

Results

On administration of 5 mg/kg of plant extract, blood glucose levels of the NIDDM rats showed 62.00 and 76.29% decrease in the blood glucose levels on day 0 and day 30 respectively. Damages caused to the kidney tissue were negligible or not seen. Serum urea and creatinine levels showed 61.49 and 70.96% decrease on day 30. LPP levels of kidney and pancreas showed 70.58 and 77.41% decrease respectively.

Conclusion

These results demonstrate significant antidiabetic and regenerative potential of the Nervilia plicata, justifying the use of plant in the indigenous system of medicine. Isolation and characterisation of the compound(s) playing pivotal role in the cure would open new vistas in the therapy of type 2 diabetes.     

Intestinal α-glucosidase inhibitory activity and toxicological evaluation of Nymphaea stellata flowers extract

Aim of the study

Nymphaea stellata willd. flowers (NSF) are used as a traditional medicine in India and Nepal to treat diabetic disease. Different works have demonstrated that NSF extract showed antihyperglycemic effect on alloxan-induced diabetic rats. In the present work we evaluated in vitro intestinal α-glucosidase inhibition as the possible mode of action of NSF extract on suppressing postprandial hyperglycemia for curing diabetic mellitus. In addition, NSF extract was studied to assess its possible acute oral toxicity and genotoxicity.

Materials and methods

Rat intestinal crude enzyme preparation and Caco-2 monolayer were used to evaluate α-glucosidase inhibitory activity of NSF extract. The main α-glucosidase inhibitors were detected by HPLC. For acute toxicity test, NSF extract was administered at doses of 2000, 5000 and 10,000 mg/kg body to three groups of 10 ICR mice each, and then clinical symptoms including mortality, clinical sign and gross findings were observed once a day for 14 days. In Ames test, histidine-dependent auxotrophic mutants of Salmonella typhimurium (strains TA97, TA98, TA100, TA102 and TA1535) were used and incubated in the presence and absence of S9 metabolic activation using NSF extract with concentrations of 150-5000 μg/plate. The chromosome aberration test was conducted with Chinese hamster lung (CHL) cells treated with NSF extract at doses of 150-5000 μg/ml in the presence and absence of S9 metabolic activation. In the in vivo mouse micronucleus assay, 9-week-old male and female ICR mice (n = 90, 25-30 g) were administered daily by oral gavage at doses of 2.5, 5.0 and 10.0 g/kg body for 1 or 2 days. Bone marrow smears were prepared from each treatment group 24 h after last administration and then polychromatic erythrocytes (PCEs) and normochromatic erythrocytes (NCEs) were identified.

Results

NSF extract showed potent rat intestinal α-glucosidase inhibitory activity for maltose hydrolysis with ED₅₀ value of 0.1 mg/ml. In Caco-2 monolayer, α-glucosidase activity for the maltose hydrolysis was down-regulated by NSF extract at a concentration of 0.05 mg/well level, showing 74% inhibition compared to the saline treated control. NSF was rich in phenol contents and the main α-glucosidase inhibitor, 1,2,3,4,6-penta-O-galloyl-β-d-glucose, was identified together with two phenolic compounds of gallic acid and corilagin. In acute toxicity test, NSF extract did not produce any toxic signs or deaths and the LD₅₀ value of this extract could be greater than 10,000 mg/kg body weight. These results of genotoxicity assessment showed that NSF extract did not cause genotoxic effects in Ames test, in the in vitro chromosomal aberration assay and in the in vivo micronucleus assay.

Conclusion

The current study shows that the extract from Nymphaea stellata flowers exhibits significant intestinal α-glucosidase inhibitory activity, without showing any acute toxicity or genotoxicity, which may be useful in suppressing postprandial hyperglycemia in diabetics. The results presented here suggest that the use of NSF in folk medicine as a natural antidiabetic treatment could be safe as well as beneficial.     

Increased sexual motivation in female rats treated with Humulus lupulus L. extract

Aim of the study

To evaluate the influence of Humulus lupulus extract on sexual behavior in female rats.

Materials and methods

Ovariectomized rats hormonally primed with estradiol benzoate (1.5 μg/rat) and progesterone (500 μg/rat) were acutely treated by oral gavage with Humulus lupulus extract dosed at 5, 10 and 25 mg/kg and then tested for partner preference and sexual receptivity.

Results

The administration of Humulus lupulus extract at the highest dose significantly increased the preference for the stimulus male during the partner preference test and the number of proceptive behaviors during the receptivity test, without affecting the lordosis response.

Conclusions

Humulus lupulus extract increased sexual motivation in hormone-primed female rats.     

Evaluation of the use of Cocos nucifera as antimalarial remedy in Malaysian folk medicine

Ethnopharmacological relevance

White flesh extract of Cocos nucifera (coconut) was studied to ascertain the ethnopharmacological standing of its antimalarial usage in Malaysian folk medicine.

Materials and methods

The crude methanol extract was investigated for phytochemical constituents and acute oral toxicity. Antimalarial activity of different extract doses of 50, 100, 200 and 400 mg/kg were investigated in vivo against Plasmodium berghei (NK65) infections in mice during early, established and residual infections. Chloroquine (20 mg/kg) and pyrimethamine (1.2 mg/kg) were used as reference drugs.

Results

The results revealed that the extract contained some phytochemical constituents and is toxicologically safe by oral administration. The extract significantly reduced the parasitaemia by the 200 and 400 mg/kg doses in the all three in vivo assessment assays. However, the extract did not significantly increase the survival time of the infected mice.

Conclusions

The observed pharmacological activities suggest that the Malaysian folkloric medicinal application of Cocos nucifera has a pharmacological basis.     

Acute and sub-chronic toxicity study of standardized extract of Fumaria indica in rodents

Ethnopharmacological relevance

Despite Fumaria indica (FI) widespread medicinal uses in the Indian traditional medicine, no systematic study of the potential toxicity of the plant has been described.

Aim of the study

To assess acute and sub-chronic toxicity of a 50% ethanolic extract of FI in mice and rats respectively.

Materials and methods

In acute toxicity study, Swiss strain albino mice of either sex were administered orally FI doses of 1, 2.5 and 5 g/kg and observed for behavioural changes and mortality, if any. In sub-chronic toxicity study, Charles Foster albino rats of either sex were administered two doses of FI i.e., 100 and 400 mg/kg, p.o. for 30 consecutive days. During 30 days of treatment, rats were observed for any change in body weight and daily food and water intake. After 30 days, rats were sacrificed for haematological, biochemical and histopathology study. Control animals were administered 0.3% carboxymethyl cellulose (CMC) suspension by oral route.

Results

There was no mortality or abnormal behaviour, observed in acute toxicity study in mice at all the three dose levels. In sub-chronic toxicity study, FI did not produce any significant change in body weight and daily food and water intake of rats when compared to vehicle treated rats. Further, haematological and biochemical parameters were also found normal. Histopathological study revealed normal architecture of kidney and liver of FI treated rats.

Conclusions

FI extract, provisionally standardized on its fumarate contents, seems to fulfill a preclinical criterion necessary for its further development as a clinically useful adaptogen.     

Anti-hypoxic activity at simulated high altitude was isolated in petroleum ether extract of Saussurea involucrata

Ethnopharmacological relevance

Rhodiola algida, Saussurea involucrata, and other herbs grown in Qinghai-Tibetan plateau have long been used to prevent and treat acute mountain sickness.

Aim of the study

To screen and identify the anti-hypoxic constituents in the herbs grown in Qinghai-Tibetan plateau of Northwestern China.

Materials and methods

The anti-hypoxic activities of 20 selected plateau herbs were examined against two positive controls, Rhodiola algida and acetazolamide, using the normobaric hypoxia model of mice. The herb with the highest activity was successively extracted with 70% ethanol, petroleum ether, chloroform, ethyl acetate and n-butanol. The extract with the highest activity was identified by comparing the survival time of mice under normobaric hypoxia condition after being subjected to different extracts. The identified extract was further tested by simulating high altitudes through an acute decompression model and a chronic decompression model for mice.

Results

The herb found to have the highest anti-hypoxic activity was Saussurea involucrate (Kar. et Kir.) Sch.-Bip, and the most effective fraction was in the petroleum ether extract. Administration of petroleum ether extract of Saussurea involucrata (PESI) to mice at 50 mg/kg significantly decreased the mortality of animals under acute decompression conditions. Changes in biochemical indicators for glycometabolism and energy metabolism, including adenosine triphosphate (ATP) content and adenosine triphosphatase (ATPase) activity in brain and cardiac muscle, lactic acid (LAC) and lactate dehydrogenase (LDH) in blood and cardiac muscles, blood sugar, and glycogen content in liver and skeletal muscle were reversed under chronic decompression conditions.

Conclusions

Saussurea involucrata (Kar. et Kir.) Sch.-Bip exhibits high anti-hypoxic activity that may be effective in preventing acute mountain sickness, and the active constituents are mainly in the petroleum ether extract.     

Antidiabetic effect of Symplocos cochinchinensis (Lour.) S. Moore. in type 2 diabetic rats

Aim

Symplocos cochinchinensis (Lour.) S. Moore. is used in Indian system of traditional medicine to treat diabetes mellitus. The present study aims to investigate the antidiabetic efficacy of the hexane extract of Symplocos cochinchinensis leaves in high fat diet-low streptozotocin (STZ) induced type 2 diabetic rats.

Materials and methods

The doses for the study were fixed based on Irwin test. The hypoglycemic effect of the hexane extract of Symplocos cochinchinensis leaves were studied in normal rats. Oral glucose and insulin tolerance tests were carried out. The antihyperglycemic effect of the hexane extract at 250 and 500 mg/kg was studied in high fat diet-low STZ induced type 2 diabetic rats for 28 days.

Results

The extracts showed no adverse effects up to 5 g/kg concentration. In hypoglycemic study, after treatment with hexane extract at 250 and 500 mg/kg the blood glucose was mildly reduced. In oral glucose tolerance test, the treatment with the hexane extract at 250 and 500 mg/kg showed a highly significant reduction of 12.07% and 23.58% in plasma glucose levels, respectively 30 min after glucose load. The insulin tolerance test also showed improved insulin sensitivity after 60 min of insulin treatment. In high fat diet-low STZ induced type 2 diabetic rats, after 28 days treatment with the hexane extract at 250 and 500 mg/kg reduced the plasma glucose level by 17.04% and 42.10%, respectively. A significant reduction in plasma insulin, plasma and hepatic total cholesterol (TC), triglycerides (TG) and free fatty acids (FFA) and a significant increase in liver glycogen were observed in treated diabetic rats.

Conclusion

This study demonstrated the potential antidiabetic property of hexane extract of Symplocos cochinchinensis leaves on type 2 diabetes mellitus, thus justifying its traditional usage.     

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.     

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.     

Anticonvulsant activity of aqueous root extract of Ficus religiosa

Ethnopharmacological relevance

Ficus religiosa Linn is frequently used for the treatment of nervous disorders among Pawara tribe of the Satpuda range, India.

Aim of the study

This study aimed to investigate the anticonvulsant activity of the aqueous aerial root extract of Ficus religiosa in chemoconvulsant-induced seizures in mice.

Materials and methods

The anticonvulsant activity of the extract (25, 50 and 100 mg/kg, p.o.) was investigated in strychnine-, pentylenetetrazole-, picrotoxin- and isoniazid-induced seizures in mice. Rat ileum and fundus strip preparations were used to study the effect of the extract on acetylcholine (Ach)- and serotonin (5-HT)-induced contractions, respectively.

Results

The extract showed no toxicity and protected the animals in the strychnine and pentylenetetrazole tests in a dose-dependent manner. Its effect in the picrotoxin and isoniazid tests, however, was less potent. The extract also exhibited dose-dependent potentiation of Ach in rat ileum but failed to potentiate the effect of 5-HT in rat fundus strip preparation.

Conclusions

The results suggest that an orally administered aqueous root extract of Ficus religiosa has dose-dependent and potent anticonvulsant activities against strychnine- and pentylenetetrazole-induced seizures. The observed activities may be ascribed to the appreciable content of zinc and magnesium in the extract.     

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.     

Kaempferia parviflora, a plant used in traditional medicine to enhance sexual performance contains large amounts of low affinity PDE5 inhibitors

Aim of the study

A number of medicinal plants are used in traditional medicine to treat erectile dysfunction. Since cyclic nucleotide PDEs inhibitors underlie several current treatments for this condition, we sought to show whether these plants might contain substantial amounts of PDE5 inhibitors.

Materials and methods

Forty one plant extracts and eight 7-methoxyflavones from Kaempferia parviflora Wall. ex Baker were screened for PDE5 and PDE6 inhibitory activities using the two-step radioactive assay. The PDE5 and PDE6 were prepared from mice lung and chicken retinas, respectively. All plant extracts were tested at 50 μg/ml whereas the pure compounds were tested at 10 μM.

Results

From forty one plant extracts tested, four showed the PDE5 inhibitory effect. The chemical constituents isolated from rhizomes of Kaempferia parviflora were further investigated on inhibitory activity against PDE5 and PDE6. The results showed that 7-methoxyflavones from this plant showed inhibition toward both enzymes. The most potent PDE5 inhibitor was 5,7-dimethoxyflavone (IC₅₀ = 10.64 ± 2.09 μM, selectivity on PDE5 over PDE6 = 3.71). Structure activity relationship showed that the methoxyl group at C-5 position of 7-methoxyflavones was necessary for PDE5 inhibition.

Conclusions

Kaempferia parviflora rhizome extract and its 7-methoxyflavone constituents had moderate inhibitory activity against PDE5. This finding provides an explanation for enhancing sexual performance in the traditional use of Kaempferia parviflora. Moreover, 5,7-dimethoxyflavones should make a useful lead compound to further develop clinically efficacious PDE5 inhibitors.     

Inhibitory effect of hydro-methanolic extract of seed of Holarrhena antidysenterica on alpha-glucosidase activity and postprandial blood glucose level in normoglycemic rat

Ethnopharmacological relevance

The present experiment was conducted to search out the effect of hydro-methanolic extract of seed of Holarrhena antidysenterica on intestinal α-glucosidase activity in dose dependent manner and on the management of postprandial hyperglycemia in starch loaded rats.

Materials and methods

Activity of intestinal α-glucosidase was measured by in vitro method. Fasting blood glucose level was determined by single touch glucometer. Total phenol and flavonoids of seed extract of Holarrhena antidysenterica were estimated using gallic acid and quercetin standard curves, respectively.

Results

The degree of elevation in blood glucose level after starch administration was significantly (p < 0.05) less by the extract in respect to the control. The said extract also inhibited α-glucosidase activity having an IC₅₀ of 0.52 mg/ml. Phytochemical study revealed that the extract is rich in phenolic compounds (60.23 mg of gallic acid equivalent/g of extract) and flavonoids (360.23 mg of quercetin equivalent/g of the extract).

Conclusion

The extract exerts its antihyperglycemic effect by retarding the carbohydrate absorption from intestine through the inhibition in α-glucosidase activity and therefore resists postprandial hyperglycemia.     

Saponin rich fractions from Polygonatum odoratum (Mill.) Druce with more potential hypoglycemic effects

Ethnopharmacological relevance

Peppers from Capsicum species (Solanaceae) are native to Central and South America, and are commonly used as food and also for a broad variety of medicinal applications.

Aim of the study

The red pepper Capsicum baccatum var. pendulum is widely consumed in Brazil, but there are few reports in the literature of studies on its chemical composition and biological properties. In this study the antioxidant and anti-inflammatory activities of Capsicum baccatum were evaluated and the total phenolic compounds and flavonoid contents were determined.

Materials and methods

The antioxidant property was assayed by scavenging abilities using DPPH and the anti-inflammatory activity was tested through the carrageenan-induced pleurisy model in mice. The total phenolic compounds and flavonoid contents were determined spectrophotometrically.

Results

The ethanolic and butanol extracts (200 mg/kg, p.o.) presented a significant anti-inflammatory activity toward carrageenan-induced pleurisy model in mice in comparison to dexamethasone (0.5 mg/kg, s.c.). Among the parameters evaluated, the treatment with these samples inhibited leukocyte migration and reduced the formation of exudate. The contents of flavonoids and total phenolic compounds could be correlated with the antioxidant and anti-inflammatory activities observed for Capsicum baccatum.

Conclusions

Our findings suggest that Capsicum baccatum contains potential antioxidant and anti-inflammatory compounds which could be tested as drug candidates against oxidative and inflammation-related pathological processes in medicinal chemistry studies.