In vivo hepatoprotective and in vitro antimicrobial potential of Ceasalpinia bonduc (Linn): Pharmacological correlation with identified phytochemicals.

The in vivo hepatoprotective potential of methanolic extract of Ceasalpinia bonduc (CBLM) has been explored against carbon tetrachloride (CCl4) induced acute liver injury in rats. Treatment of plant extract on CCl4 intoxicated liver significantly reduced the hepatoxicity, along with serum enzymes GPT and GOT. To explore the chemical constituents from CBLM extract, it was fractionated into non-polar to moderately polar fractions (CBLM-H, CBLM-HEt, CBLM-Et, CBLM-EtM, CBLM-M) and subjected to GC/GC-MS analysis. Altogether twenty seven (~71%) phytochemicals were identified from different fractions by using Electronic Mass Spectral Library GC-MS (NIST 20). Out of which twenty one are first time reported from Ceasalpinia bonduc, fourteen from genus Caesalpinia and ten from family Fabaceae. The identified phytochemicals 2-ethyl-2-hydroxy-1,3-dimethylcyclopentanecarboxylic acid, ethyl ester (21) and 1,3,5-triazine-2,4-diamine,6-hydroxy-N,N-dicyclohexyl (23) are first time identified as plant metabolites. To explore the antimicrobial potential four strains of Gram-positive and eight strains of Gram-negative bacteria were used along with pure cultures of five saprophytic fungus (molds) and two strains of yeast were utilized. CBLM-H and CBLM-HEt were exhibited praiseworthy antimicrobial potential. CBLM-H showed complete growth inhibition of P. mirabilis and V. cholerae at the concentration of 0.1g/mL while CBLM-HEt at 0.05g/mL halted the growth of S. aureus.

Neurochemical and behavioral effects of Nigella sativa and Olea europaea oil in rats.

OBJECTIVES: In the last few decades, therapeutic uses of medicinal compounds present in food as a normal constituent has risen substantially, largely because of their fewer side effects and adequate efficacy. This study is designed to investigate a role of brain serotonin (5-HT) and dopamine (DA) in the potential nootropic, anxiolytic, and other beneficial effects of Nigella sativa (NS) and Olea europaea (OE) oil in rat models. METHODS: Animals were treated with NS and OE oil orally at doses of 0.1 ml/kg and 0.25 ml/kg for 5 weeks. Food intake and body weight change, anxiety-like effects in elevated plus maze and activity in a novel and familiar environment were monitored weekly. Effects on learning and memory after 5 weeks treatment were monitored using Morris water maze test. Neurochemical analysis was carried using HPLC-ECD method. RESULTS: NS and OE oil administration enhanced learning and memory in Morris water maze test and the effects were greater in NS than OE oil-treated animals. Low dose of OE oil increased exploration in an open field, higher dose of OE oil and both doses of NS oil produced no consistent effect on open field exploration. Effects of both oils on anxiety-like behavior, food and water intake, and activity in activity box were either not consistent or did not occur. The treatment increased homovanillic acid (HVA). 5-HT levels increased in high dose of NS oil and low dose of OE oil-treated groups. Low dose NS oil decreased 5-HT. DISCUSSION: The present study suggests that active components in NS and OE oil may prove useful in treating impaired cognition. OE oil may produce psychostimulant-like effect. Modulation of DA and serotonin neurotransmission seems important in the pharmacological effect of these oils.

Antidepressant-like deliverables from the sea: evidence on the efficacy of three different brown seaweeds via involvement of monoaminergic system.

Brown seaweeds exhibit several health benefits in treating and managing wide array of ailments. In this study, the antidepressant-like effect of methaolic extracts from Sargassum swartzii (SS), Stoechospermum marginatum (SM), and Nizamuddinia zanardinii (NZ) was examined in forced swimming test (FST), in rats. Oral administration of SS, SM, and NZ extract (30-60 mg/kg) exhibited antidepressant-like activity in FST by reducing immobility time as compared to control group, without inducing significant change in ambulatory behavior in open field test. In order to evaluate the involvement of monoaminergic system, rats were pretreated with the inhibitor of brain serotonin stores p-chlorophenylalanin (PCPA), dopamine (SCH23390 and sulpiride), and adrenoceptor (prazosin and propranolol) antagonists. Rats receiving treatment for 28 days were decapitated and brains were analyzed for monoamine levels. It may be concluded that the extracts of SS, SM, and NZ produces antidepressant-like activity via modulation of brain monoaminergic system in a rat model.

Effect of dichloromethane fraction of Areca catechu nut on monoamines associated behaviors and tyramine pressor sensitivity in rodents.

The current study was aimed at investigating the effect of Areca catechu nut dichloromethane fraction (7 mg/kg) on monoamines (serotonin and dopamine) modulation (5-hydroxytryptophan-induced tremors and phenylethylamine-induced stereotypes) and its interaction with tyramine (cheese effect). The dichloromethane fraction caused pronounced increase in 5-HTP-induced tremors (50%) with negligible PEA-induced stereotypes (20%). Additionally, it did not produce a significant increase in the tyramine pressor effects. These results suggest that the dichloromethane fraction of A. catechu nut primarily elevates serotonin levels (probably via monoamine oxidase A inhibition) and does not induce cheese effect.

Potential antidepressant activity of Areca catechu nut via elevation of serotonin and noradrenaline in the hippocampus of rats.

The current study was aimed at investigating the potential antidepressant activity of Areca catechu nut ethanol extract and its various fractions using behavioral (acute and sub-chronic forced swim tests) and biochemical (monoamines and their metabolite levels using high performance liquid chromatography) tests. The areca nut ethanol extract and its aqueous fraction exhibited antidepressant activity in both acute and sub-chronic forced swim tests (IC50 ~ 50 and 20 mg/kg, respectively), which was further confirmed by unaltered locomotor (horizontal and vertical) activities of rats in the activity cage. Phytochemical analysis revealed that saponins of areca nut may be the active component in its antidepressant action. The rats treated sub-chronically with areca nut extract displayed toxic effects, whereas its active aqueous fraction was non-toxic, indicating the presence of different constituents for antidepressant and toxic effects. In the hippocampus of rats, the areca nut extract (50 mg/kg) and aqueous fraction (20 mg/kg) caused a significant elevation of serotonin (around 35%) and noradrenaline (around 30%) compared with the control (261 ± 25 and 512 ± 29 ng/g, respectively). In conclusion, the areca nut possesses potential antidepressant effect via the elevation of serotonin and noradrenaline.

Biochemical and haematological evaluation of repeated dose exposure of male Wistar rats to an ethanolic extract of Artemisia annua.

Artemisia annua is widely used for the treatment of malaria and other disorders. In a previous study, the artemisinin concentration in the dry leaves of A. annua grown under humid tropical conditions was determined to be 1.098% using reversed phase high performance liquid chromatography. In the current study, biochemical and haematological evaluations of ethanolic leaf extracts derived from such plants (EAA) were carried out in 20 male Wistar rats. Rats were divided into four study groups of saline-treated (control) and test groups exposed orally to graded doses of EAA for 28 days. The results showed that the liver function and haematological indices, and testosterone levels were not adversely affected. High density lipoprotein -cholesterol was reduced at 100 mg/kg of EAA, atherogenic index as well as low density lipoprotein -cholesterol was raised, and glucose concentration was reduced significantly at the 100 and 200 mg/kg of EAA (p < 0.05). In addition to serving as a possible antidiabetic agent, EAA may not predispose users to hepatotoxicity, haematotoxicity and testicular toxicity. However, due to the possible risk of atherosclerosis, we advise that the plant extract should be taken with caution in people with atherosclerotic condition.

A safety assessment of the antimalarial herb Artemisia annua during pregnancy in Wistar rats.

Artemisia annua is a Chinese antimalarial herb that has been used for more than 2000 years. The maternal and foetal safety of the ethanolic leaf extract of therapeutically active Artemisia annua (EAA), with previously determined artemisinin yield of 1.098% was evaluated in Wistar rats. Twenty pregnant rats, divided into four study groups of saline treated (control), and test groups administered orally with 100, 200 and 300 mg/kg body weights of EAA, respectively, from gestation days (GD) 8 to 19. Following overnight fast, animals were sacrificed on GD 20, and maternal blood was collected to evaluate biochemical and haematological markers. Foetuses were carefully removed, weighed, and observed for any possible malformation. Biochemical and haematological studies revealed that EAA did not result in maternal hepatotoxicity, haematotoxicity, and hyperlipidemia. While litter size significantly decreased (p < 0.05) at 100 mg/kg EAA, maternal estrogen levels decreased in all the EAA-treated groups. Non-viable (21%) and malformed (31%) foetuses were observed at the 300 mg/kg dose of EAA, which implies that although consumption of the leaf extract may not predispose users to hepatotoxicity, haematotoxicity, and hyperlipidemia, it should be taken with caution during pregnancy due to possible risk of embryotoxicity at concentrations higher than the therapeutic dose.

Comparison of monoamine reuptake inhibitors for the immobility time and serotonin levels in the hippocampus and plasma of sub-chronically forced swim stressed rats.

The current study was aimed at comparing the behavioral and biochemical (5-hydroxytryptamine and 5-hydroxyindoleacetic acid levels) effects of monoamine reuptake inhibitors (fluoxetine, venlafaxine and imipramine) in sub-chronically forced swim stressed rats. At the given doses of 10, 20 and 30 mg/kg, among aforesaid antidepressants, the imipramine treatment alone caused significant decline in the immobility time of rats (IC(50) 20 mg/kg). In the hippocampus of rats, the imipramine treatment caused significant elevation of 5-hydroxytryptamine (5-HT) whereas, the fluoxetine and venlafaxine elicited significant increase in 5-hydroxyindoleacetic acid (5-HIAA) levels. Likewise, in the plasma of rats, the imipramine treatment significantly increased the 5-HIAA levels whereas, the fluoxetine and venlafaxine treatment significantly elevate the 5-HT levels. It can therefore be inferred that the imipramine did not act like other monoamine reuptake inhibitors in biochemical study, which could possibly underlie its ability to be detected in forced swim test (behavioral study). Moreover, the re-uptake inhibition of 5-HT is not accountable for the antidepressant action exhibited in forced swim test.

Bioassay-guided isolation of antioxidant agents with analgesic properties from flowers of Tagetes patula.

CONTEXT: Tagetes patula L. is one of the French marigold group of the Asteraceae family. It is recognized in folklore for its medicinal and pesticidal properties. OBJECTIVE: In search of more effective, but non-toxic compounds with antioxidative potential led to the bioassay guided isolation studies on the extracts of T. patula. MATERIALS AND METHODS: The bioassay on Tagetes patula flowers were carried out guided by in vitro antioxidant activity using DPPH assay. A minor but proven plant constituent methyl protocatechuate (1) was isolated by column chromatography, while patuletin (2) and patulitrin (3) obtained in bulk by employing solvent partition of methanol extract. Derivatization of patuletin into benzoyl, cinnamoyl and methyl was conducted to establish the structure activity relationship (SAR). Analgesic activity of compound 2 was evaluated using acetic acid-induced writhing test and hot-plate test in mice. The toxicity of methanol extract and compound 2 were also determined. RESULTS: Polar extracts, fractions and phases demonstrated better antioxidant activity. The synthetic methyl protocatechuate (1) showed IC(50) value of 2.8 ± 0.2 µg/mL, whereas patuletin (2) (IC(50) = 4.3 ± 0.25 µg/mL) was comparable to quercetin and rutin but significantly better than patulitrin (3) (IC(50) = 10.17 ± 1.16 µg/mL). Toxicity test for the methanol extract and compound 2 did not elicit any behavioral changes or cause mortality in mice. Compound 2 also demonstrated mild analgesic property. DISCUSSION AND CONCLUSION: These findings demonstrate that the plant polar extracts and fractions possess significant antioxidant property with non-toxic effect. Compound 1 is a genuine plant constituent of T. patula.

Aegiceras corniculatum extract suppresses initial and late phases of inflammation in rat paw and attenuates the production of eicosanoids in rat neutrophils and human platelets.

AIM OF THE STUDY: The present study is designed to explore the anti-inflammatory potential of Aegiceras corniculatum Linn. Blanco stems extracts and their mechanism of action against various pro-inflammatory mediators and to validate its traditional use against inflammatory diseases. MATERIALS AND METHODS: Rat paw edema and peritonitis models were employed for in vivo studies. For in vitro studies human platelets and rat neutrophils were stimulated with Ca(2+)-ionophore A23187 leading to the production of various pro-inflammatory metabolites, i.e., 12-HTT, 12-HETE and LTB(4) and 5-HETE which were quantified by HPLC. RESULTS: The highly polar methanol extract (100mg/kg) caused approximately 90% reduction in the carrageenan- and prostaglandin E2-induced paw edema in rats. It also caused the inhibition of cycloxygenase-1 metabolite, 12-HHT (IC(50) 41.1+/-1.5microg/ml) with a concomitant rise in 12-lipoxygenase metabolite, 12-HETE in A23187 stimulated human platelets. Conversely, the non-polar hexane extract attenuated (IC(50) 0.36+/-0.12microg/ml) 12-HETE formation with a parallel rise in 12-HHT, thereby displaying a selectivity towards 12-lipoxygenase. Non-polar hexane extract also antagonized the production of 5-lipoxygenase metabolites, i.e., leukotriene B(4) and 5-HETE in the rat neutrophils. Furthermore, ethyl acetate extract inhibited both COX and 5-LOX with a marked decline in the production of 12-HHT (IC(50) 0.08+/-0.002microg/ml) and LTB(4) (IC(50) 0.86+/-0.03microg/ml), respectively. The anti-inflammatory effect of hexane and ethyl acetate extracts was also reflected by the diminution of carrageenan-induced cell infiltration in rat peritoneum. Additionally, plant extracts caused approximately 60% suppression in dextran-induced paw edema implying that they also ameliorate histamine and serotonin release. CONCLUSION: Hexane, ethyl acetate and methanol extracts derived from Aegiceras corniculatum possess significant anti-inflammatory activity via multiple mechanisms and validate their traditional use against inflammation-related diseases.

A study on antioxidant, free radical scavenging, anti-inflammatory and hepatoprotective actions of Aegiceras corniculatum (stem) extracts.

AIM OF THE STUDY: The present study was conducted to evaluate the antioxidant, anti-inflammatory and hepatoprotective potential of Aegiceras corniculatum Linn. Blanco (Aegicerataceae). METHODS AND RESULTS: The n-hexane, ethyl acetate and methanol extracts, derived from Aegiceras corniculatum stems, scavenged superoxide anions (O2*) and hydroxyl radicals (*OH) in nitro blue tetrazolium reduction and deoxyribose degradation assays, respectively. All the extracts inhibited the process of lipid peroxidation at its initiation step. Additionally, in rat liver microsomes n-hexane and ethyl acetate extracts also caused termination of radical chain reaction supporting their scavenging action towards lipid peroxy radicals (LOO*). Moreover, increased production of O2* in human neutrophils, stimulated by phorbol-12-myristate-13-acetate (PMA) and/or opsonized zymosan were also suppressed (IC50 approximately 3-20 microg/mL). Thereby, revealing the ability of plant extracts to antagonize the oxidative stress via interference with NADPH oxidase metabolic pathway. These in vitro results coincide with the reduction in the glucose oxidase-induced paw edema in mice in the presence of ethyl acetate and methanol extracts (10, 50, and 100mg/kg, i.p.). Plant extracts (250, 500 and 1000 mg/kg, p.o.) also significantly protected the carbon tetrachloride (CCl4)-induced oxidative tissue injury in rat liver. This was reflected by a approximately 60% decline in the levels of serum aminotransferase enzymes. CONCLUSION: Aegiceras corniculatum extracts found to possess pronounced antioxidant effect that may be at least in part related to its anti-inflammatory and hepatoprotective activities. This study provides a scientific basis for the ethnomedical claims that Aegiceras corniculatum is effective against inflammation and liver injury.

Sargassum swartzii extracts ameliorate memory functions by neurochemical modulation in a rat model.

Recently, considerable attention has been paid to drug exploration from natural sources for treating memory loss, a major manifestation of various neurodegenerative diseases. Increasing evidences implicate brain serotonin metabolism in learning and memory, supporting the notion that targeting 5-HT (5-hydroxytryptamine) and its receptors would be beneficial in the treatment of cognitive disorders. In the present study, behavioral and neurochemical effects were examined following administration of Sargassum swartzii extracts in albino Wistar rats. Increase in spatial working memory and recognition memory was exhibited by the seaweed-treated rats as compared to controls. Plasma tryptophan, brain 5-HT, and 5-hydroxyindoleacetic acid levels were measured using HPLC-ECD, and a significant increase in brain 5-HT metabolism was observed in the seaweed-treated rats. The increase in memory functions following repeated administration of S. swartzii extracts is suggested to be due to the increased serotonergic neurotransmission in the brain of seaweed-treated rats.

Anti-inflammatory activity of Bacopa monniera in rodents.

The ethanol extract of Bacopa monniera (Scrophulariaceae) exhibited marked anti-inflammatory activity against carrageenan-induced paw edema in mice and rats, an acute inflammatory model. To assess the possible mechanism of anti-inflammatory action against carrageenan, the ethanol extract was treated with chemical mediators (histamine, serotonin, bradykinin, prostaglandin E(2) and arachidonic acid)-induced edema in rats. The extract selectively inhibited prostaglandin E(2)-induced inflammation. Thus, it may be inferred that B. monniera possesses significant anti-inflammatory activity that may well be relevant for its effectiveness in the healing of various inflammatory conditions in traditional medicine.

Genetic instability in EBV-transformed lymphoblastoid cell lines.

Epstein Barr virus (EBV)-transformed lymphoblastoid cell lines are commonly used to provide an inexhaustible supply of DNA. We examined microsatellite instability in these cell lines in 35 individuals where DNA was available from the original blood samples and from cultured cell lines. Mutations were observed in 0.3% of the analyses, thus providing a quantitative measure of somatic mutation rate.

Evaluation of Alstonia scholaris leaves for broncho-vasodilatory activity.

The present study demonstrates that the ethanol extract of Alstonia scholaris (Apocynaceae) leaves induced pronounced bronchodilatory activity in anaesthetized rats with the probable involvement of prostaglandins. However, in vitro preparations of guinea-pig trachea did not confirm this property, indicating that bronchodilation is not due to the direct tracheal smooth muscle relaxation. The vasodilatory activity of the extract was independent of adrenergic or muscarinic receptors or prostaglandins but was mainly via endothelial-derived relaxing factor, nitric oxide. The extract inhibited the spontaneous movements of rabbit jejunum and contractile effects of acetylcholine and histamine on guinea-pig ileum. Additionally, the extract caused marked reduction of barium chloride-, potassium chloride- and calcium chloride-induced contraction on guinea-pig ileum and pulmonary artery, implying a direct interference of plant extract with the influx of calcium ions into cells. However, the extract has no detectable effect on mobilization of intracellular calcium. These results coupled with the in vivo effects of ethanol extract reveal that the Alstonia scholaris leaves possess broncho-vasodilatory activity mediated presumably by prostaglandins, calcium antagonism and endothelium-derived relaxing factor(s).

Antinociceptive diterpene from Euphorbia decipiens.

One myrsinol-type diterpene ester (1) isolated from Euphorbia decipiens was evaluated for analgesic activity in the acetic acid induced writhing test in mice. Different dose (5-20 mg/kg i.p.) of the compound showed significant antinociceptive activity, which was comparable to standard analgesic drugs, aspirin and ibuprofen (100 mg/kg i.p.).

In-vitro assessment of cytotoxicity of halloysite nanotubes against HepG2, HCT116 and human peripheral blood lymphocytes.

Halloysite is a clay mineral with chemical similarity to kaolin, a pharmaceutical ingredient. It consists of mainly aluminosilicate nanotubular particles in the size range of ∼ 200-1000 nm. Many studies have tried to empirically explore this novel clay for its potential in drug delivery systems but no work has yet studied its cytotoxicity from the perspective of oral drug delivery system. In this study, the halloysite nanotubes (HNTs) were subjected to size distribution analyses, which reveal more than 50% of nanotubes in the size range of 500 nm and rest mainly in the sub micrometer range. HNTs were then evaluated for in-vitro cytotoxicity against HCT116 (colorectal carcinoma) and HepG2 (hepatocellular carcinoma) cells which represent the earliest entry point and the first accumulating organ, respectively, for nanoparticles en-route to systemic circulation after oral delivery. Moreover, HNTs were tested for their cytogenetic toxicity against human peripheral blood lymphocytes. Both these results collectively indicated that HNTs are generally safe at practical concentrations of excipients for oral dosage forms.

Broncho-vasodilatory activity of fractions and pure constituents isolated from Bacopa monniera.

The present study demonstrates that various fractions and sub-fractions isolated from Bacopa monniera produced significant inhibition of carbachol-induced bronchoconstriction, hypotension and bradycardia in anaesthetized rats. All these showed more potency towards inhibition of tracheal pressure compared to either blood pressure or heart rate. The sub-sub fraction and compound 1 caused greater inhibition of tracheal pressure and heart rate compared to blood pressure. Thus, overall bioassay-directed fractionation of B. monniera improved the bronchodilatory activity in various fractions and compound 1 (2-219x) in anaesthetized rats. In vitro, the KCl-induced contraction was equally inhibited by crude extract, petroleum ether and methanol fractions on trachea suggesting bronchodilatory activity remained the same in fractions. On pulmonary artery petroleum ether, dichloromethane and methanol fractions produced 2-2.6 times more vasodilatation compared to crude extract of B. monniera. Subsequent sub-fractions failed to show the existence of broncho-vasodilatory activity, however, the CHCl(3)/MeOH sub-fraction significantly reduced the acetylcholine-induced contraction on ileum. Both the methanol fraction and CHCl(3)/MeOH sub-fraction caused marked reduction of barium chloride-, potassium chloride- and calcium chloride-induced contraction on guinea-pig ileum, indicating their interference with Ca(2+) ion movement. Thus, it may be concluded that various fractions derived from B. monniera possess broncho-vasodilatory activity, which is attributed mainly to inhibition of calcium ions.

Antinociceptive activity of methanolic extracts of St. John's Wort (Hypericum perforatum) preparation.

The analgesic effect of St. John's Wort plant preparation was evaluated using chemically induced pain models in adult albino mice. The intraperitoneal (i.p.) administration of 30-100 mg/kg of the preparation produced significant analgesic effect (75%) in acetic acid induced writhing and formalin licking tests. Based on its inhibitory effects on acetic acid induced writhes and both phases of the formalin test, the plant preparation seems to have central and peripheral analgesic properties. Its effect was found twice more potent than ibuprofen (100 mg/kg), a standard reference drug, in the acetic acid induced writhing test.

Artemisia annua as a possible contraceptive agent: a clue from mammalian rat model.

In a previous study, we evaluated the maternal and fetal safety of antimalarial herb Artemisia annua with artemisinin yield of 1.09%. Here, we attempted to ascertain the contraceptive claim of A. annua. Sexually matured female Wistar rats (180-220 g) were allotted into four study groups of six rats each. The control group received normal saline, while the A. annua-treated groups received 100, 200 and 300 mg/kg of A. annua for 2 weeks, followed by mating with proven fertile males (1:1). The rats were allowed to carry the pregnancy to term. At birth and weaning periods, selected reproductive outcome and fertility indices were determined. The results showed that A. annua significantly reduced litter size, reproductive outcome and fertility indices compared with the control (p <  0.05). These results imply that A. annua could serve as a prospective contraceptive agent in addition to its antimalarial activity.