Anti-inflammatory, analgesic and antipyretic properties of Thespesia populnea Soland ex. Correa seed extracts and its fractions in animal models

Ethnopharmacological relevance

Thespesia populnea Sol. ex Correa (Malvaceae) is commonly known as “Indian tulip tree”. The plant has been used as an astringent, antibacterial, anti-inflammatory, antinociceptive and hepatoprotective in Indian system of traditional medicine.

Materials and methods

Thespesia populnea seeds were successively extracted by soxhlet extraction using petroleum ether (40-60 °C) (TPO) and ethanol (TPE). Unsaponifiable matter (TPOUM) and fatty acids were separated from seed oil. A GC-MS analysis of fatty acid methyl esters was carried out. Ethanolic extract was fractionated using CHCl₃, EtOAc, n-BuOH and H₂O. Acute arthritis was induced by sub-plantar injection of carrageenan into the left hind paw of rats. The paw volume was measured using plethysmometer. Analgesic activity was assessed by heat induced pains (tail immersion model) and antipyretic activity assessed using brewers yeast-induced pyrexia model.

Results

Oral administration of TPO and TPE at 200 and 400 mg/kg b.w. and tested fractions at 200 mg/kg significantly reduced carrageenan induced paw edema and brewers yeast-induced pyrexia. In tail immersion method also extracts and fractions showed significant analgesic activity. Amongst all fractions EtOAc showed most significant results. TPOUM showed significant anti-inflammatory and analgesic activity. GC-MS analyses showed the presence of fourteen fatty acids, predominant fatty acids were palmitic and stearic acid.

Conclusion

These results highlighted anti-inflammatory, analgesic and antipyretic effects of Thespesia populnea seeds.     

Anti-inflammatory effect of a methanolic extract of leaves of Dregea volubilis

Aim of study

The aim of this study was to establish the anti-inflammatory activity of the methanolic extract of Dregea volubilis leaves (MEDV) with its fractions and to delineate the possible mechanism of action for MEDV.

Materials and methods

The anti-inflammatory activities of MEDV along with its petroleum ether and chloroform fractions were evaluated in a carrageenan induced model of acute inflammation. The effect of MEDV on lipopolysaccharide induced production of nitric oxide (NO) in macrophages was also studied.

Results

MEDV (100, 200 and 400 mg/kg body weight) significantly reduced carrageenan induced paw edema; chloroform fraction was most potent (66%, p < 0.001). MEDV was non-toxic up to 125 μg/ml in mouse peritoneal macrophages wherein it (0-100 μg/ml) reduced lipopolysaccharide induced NO production.

Conclusion

MEDV possesses significant anti-inflammatory activity. Chloroform fraction of MEDV showed best anti-inflammatory activity.     

Anti-ulcer activity of Cyrtocarpa procera analogous to that of Amphipterygium adstringens, both assayed on the experimental gastric injury in rats

Ethnopharmacological relevance

The bark of Amphipterygium adstringens (Aa) is commonly mixed or adulterated with the bark of Cyrtocarpa procera (Cp) and sold in Mexican markets. Aa is a well known species in Mexico used as decoction to relieve ulcers. Scientific reports reinforcing the anti-ulcer activity of Aa have been previously described, but those describing the anti-ulcer properties of Cp as a substitute for Aa in folk medicine are scarce.

Aim of the study

To investigate anatomical and phytochemical differences between these species, as well as to assess the anti-ulcer effect of Cp extracts in comparison to the Aa extracts.

Material and methods

Anatomical micro-technique and physical and spectroscopic data were used to analyze differences between Cp and Aa. Regard to the pharmacological activity, it was assessed by using the ethanol-induced gastric damage model in rats.

Results

Whereas the bark anatomy of Aa was characterized by vertical canals in the periderm and the rare occurrence of fibers in its phloem, a periderm without vertical canals and abundant fibers in the phloem were distinctive features of Cp. Phytochemical analysis allowed the identification of tirucallane, masticadienonic and 3α-hydroxymasticadienonic acids as major components in Aa, while β-amyrin and β-sitosterol were obtained from Cp. Gastric lesions observed in the control group decreased in the presence of 100 mg/kg of hexane, ethyl acetate and methanol extracts from the normal or regenerated bark of Cp, thus resembling the anti-ulcer effect of Aa. Nevertheless, major anti-ulcer potency was observed with the most active methanol extract from Cp obtained from normal [the effective dose fifty ED₅₀ = 45.54 mg/kg] or regenerated (ED₅₀ = 36.68 mg/kg) bark in comparison to Aa (ED₅₀ = 115.64 mg/kg).

Conclusion

Chemical and anatomical differences were found between these species, but since the anti-ulcer activity of Cp is similar to that shown by Aa our results reinforce the use of both species for the relief of gastric ulcer in folk medicine.     

Hepatoprotective and antioxidant effects of the methanolic extract from Halenia elliptica

Aims of study

Halenia elliptica, a medicinal herb of Tibetan origin, was commonly used in folk medicine to treat hepatitis. The aim of the present study is to evaluate the hepatoprotective and antioxidant activity of Halenia elliptica against experimentally induced liver injury.

Materials and methods

The antioxidant property of methanolic extract (ME) of Halenia elliptica was investigated by employing various established in vitro systems. The ME of Halenia elliptica was studied here for its hepatoprotective effects against CCl₄-induced liver toxicity in rats. Activity was measured by monitoring the levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and total bilirubin.

Results

The ME possessed strong antioxidant activity in vitro. The results of CCl₄-induced liver toxicity experiment showed that rats treated with the ME of Halenia elliptica (100 mg/kg and 200 mg/kg), and also the standard treatment, silymarin (50 mg/kg), showed a significant decrease in ALT, AST, ALP, and total bilirubin levels, which were all elevated in the CCl₄ group (p < 0.01). The results observed after administration of 100 mg/kg ME were comparable to those of silymarin at 50 mg/kg (p > 0.05). The ME did not show any mortality at doses up to 2000 g/kg body weight.

Conclusion

These results seem to support the traditional use of Halenia elliptica in pathologies involving hepatotoxicity, and the possible mechanism of this activity may be due to strong free radical-scavenging and antioxidant activities of ME.     

Reproductive toxicity of Rhizoma Sparganii (Sparganium stoloniferum Buch.-Ham.) in mice: mechanisms of anti-angiogenesis and anti-estrogen pharmacologic activities

Ethnopharmacological relevance

Indications and preliminary studies of Rhizoma Sparganii (RS) suggest its pharmacological mechanism is involved with endocrine/angiogenesis functions. We therefore studied its potential toxicity on reproduction in mice.

Materials and methods

Reproductive toxicity of 100, 200 and 400 mg/kg RS extract were studied in pregnant ICR mice and its offspring. The embryos’ fibroblast growth factor-1 (FGF-1), vascular endothelial growth factor (VEGF) and estrogen receptor-α (ER-α) were evaluated as targets of endocrine/angiogenesis by immunohistochemical test.

Results

The offspring of treated mice (100, 200 and 400 mg/kg RS extract) during their pregnancy had various pathological conditions, suggesting an abnormal FGF signaling phenomenon during pregnancy. Embryos from the 400 mg/kg group had significantly depressed levels of FGF-1 (P < 0.01) and VEGF (P < 0.05) expression levels as compared to controls by immunohistochemical test. Dysplasia in the heart (12.9%), craniofacial region (18.3%) and vertebrae (32.5%) presented in embryos of the 400 mg/kg group. Furthermore, the ER-α expression was inversely proportional to FGF-1 levels in the same embryo (P < 0.01).

Conclusions

These results implicate a FGF signaling abnormality in vivo and indicate that RS has anti-angiogenesis and anti-estrogen toxicity effects in pregnant rodents.     

Evaluation of anti-migraine potential of Areca catechu to prevent nitroglycerin-induced delayed inflammation in rat meninges: possible involvement of NOS inhibition

Ethnopharmacological relevance

Areca catechu nut extract is a popular folk remedy for the treatment of migraine in Kerala and Tamil Nadu states of India.

Aim of the study

In order to prove the claimed utilization of plant, the effect of hydroalcoholic extract of Areca catechu nut (ANE) was investigated in nitroglycerine induced inflammation in rat meninges. In these models infusion of nitric oxide donor glyceryl trinitrate (GTN) produces augmented plasma protein extravasation (PPE) in dura mater, provides an important substrate for the development of migraine in rats.

Materials and methods

The effect on plasma protein extravasation was assessed in both the models of intravenous and topical GTN application following oral administration of ANE (250 mg/kg and 500 mg/kg) in both curative and preventive treatment and compared with that of control positive. The l-NAME (15 mg/kg, i.v.) was used as reference standard. Plasma protein extravasation was measured using fluorescein as marker and was measured using a Perkin-Elmer LS-30 luminescence spectrometer.

Results

Expression of iNOS in the spleen after intravenous injection produced PPE into the dura mater in control positive group was significantly (P < 0.01) reduced to 1.553 ± 0.02499 and 1.398 ± 0.01887 by preventive treatment with ANE at the dose of 250 and 500 mg/kg, orally, respectively. The extravasation produced by topical GTN due to expression of iNOS in dural macrophages was also reduced to 1.555 ± 0.03384 and 1.425 ± 0.01204 by preventive treatment with ANE at the dose of 250 and 500 mg/kg, orally, respectively. While ANE do not showed any significant results in curative treatment in both the models of i.v. and topical GTN application.

Conclusion

These findings collectively indicate that the extract exhibited significant inhibition of iNOS, which may be the probable mechanism for its anti-migraine activity, providing evidence, at least in part, for its folkloric use.     

Identification of trans-tiliroside as active principle with anti-hyperglycemic, anti-hyperlipidemic and antioxidant effects from Potentilla chinesis

Aim of the study

The present study was carried out to isolate and identify trans-tiliroside as principal compound with anti-hyperglycemic, anti-hyperlipidemic and antioxidant effects from Potentilla chinesis.

Materials and methods

A bioactive compound, trans-tiliroside was isolated from the ethanol extract of Potentilla chinesis and its administration dose was optimized and patented. The normal, alloxan-induced diabetic mice and streptozotocin-induced diabetic rats were used to evaluate the anti-hyperglycemic, anti-hyperlipidemic and antioxidant effects of trans-tiliroside from Potentilla chinesis. Biochemical parameters were assayed in blood samples of different groups of alloxan-induced diabetic mice and streptozotocin-induced diabetic rats.

Results

The level of fasting serum glucose levels, triglycerides (TG) and total cholesterol (TC) in alloxan-induced diabetic mice were significantly decrease after daily oral administration of trans-tiliroside in normal and diabetic mice at doses of 0.4, 0.8 and 1.6 mg/kg/day, for 15 days. Blood glucose level was significant decrease in STZ induced diabetic rats by trans-tiliroside (1.2 and 0.3 mg/kg body weight for 10 weeks). The content of TC, low density lipoprotein (LDL-C) and TG levels were decreased and high density lipoprotein (HDL-C) content was increased, so lipid metabolism was improved. Moreover, trans-tiliroside revealed antioxidant activity as shown by increased activities of superoxide dismutase (SOD), decreased level of malondialdehyde (MDA) in diabetic rats. Histological morphology examination showed that the trans-tiliroside restored the damage of pancreas tissues in rats with diabetes mellitus.

Conclusion

Trans-tiliroside, a constituent from Potentilla chinesis, revealed significant anti-hyperglycemic, anti-hyperlipidemic and antioxidant activities.     

An herbal fruit, Amomum xanthoides, ameliorates thioacetamide-induced hepatic fibrosis in rat via antioxidative system

Aim of the study

Amomum xanthoides is a well-known traditional herbal medicine mainly for diverse digestive system disorders in Asia for a long time. In the present study, we investigate the effects and action mechanism of methanol fraction of Amomum xanthoides (MFAX) on thioacetamide (TAA)-induced liver fibrosis in rat model.

Materials and methods

TAA (200 mg/kg, ip on twice a week for 14 weeks) treated rats were orally administered with MFAX (25, 50 or 100 mg/kg) once a day from the 7th week until 14th week.

Result

Significantly elevated serum bilirubin, liver tissue hydroxyproline and malondialdehyde (MDA) in liver fibrosis were ameliorated by MFAX treatment. Further, MFAX treatment attenuated the reactive oxygen species (ROS) levels and restored glutathione (GSH) content and glutathione-peroxidase (GPx) activity. Histopathological data showed that MFAX treatment inhibited collagen accumulation and activation of hepatocyte stellate cells (HSCs) in the liver tissue. Compared to the TAA group, activation of inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-α), transforming growth factor beta (TGF-β), platelet-derived growth factor beta (PDGF-β) mRNAs and the level of pro-fibrotic cytokines PDGF-β and connective tissue growth factor (CTGF) in the liver tissue were attenuated in MFAX treated groups.

Conclusion

The above evidences collectively indicate that MFAX is a potential herb which can be used as an anti-hepatofibrotic remedy.     

Normal and delayed wound healing is improved by sesamol, an active constituent of Sesamum indicum (L.) in albino rats

Ethno-pharmacological relevance

The seeds of Sesamum indicum Linn. (Pedaliaceae) has been used traditionally for the treatment of wounds in Buldhana district of Maharashtra state. Sesamol is the main anti-oxidative constituent contained mainly in the processed sesame seed oil which has not been explored scientifically for its wound healing activity.

Aim of the study

To investigate the influence of sesamol (SM) on wound repair, both in normal and dexamethasone (DM) delayed healing processes in albino rats.

Materials and methods

Incision, excision and dead space wounds were inflicted on albino rats (180-220 g) of either sex, under ketamine anaesthesia. Group I served as control, group II received SM 50 mg/kg i.p., group III was treated with dexamethasone (DM) i.m. (0.17 mg/kg) and SM + DM was given to group IV. The tensile strength, wound contraction, hydroxyproline, lysyl oxidase and total RNA and DNA levels (in granulation tissue) were measured.

Results

The tensile strength significantly (p < 0.05) increased with SM at 471.40 ± 14.66 g when compared to control at 300.60 ± 9.16 g in normal and DM suppressed healing. No significant change was observed in duration of wound contraction and lysyl oxidase when compared to control at 2.98 ± 0.10 mg. SM treated rats showed a significant (p < 0.05) rise in hydroxyproline levels at 6.45 ± 0.45 mg when compared to control at 1.75 ± 0.20 mg.

Conclusion

These results indicate that sesamol could be a promising drug in normal as well as delayed wound healing processes.     

Inhibitory effects of processed Aconiti tuber on morphine-induced conditioned place preference in rats

Aim of the study

Our previous studies indicated that processed Aconiti tuber (PAT), a traditional Chinese herbal medicine, had antinociceptive effects and inhibitory effects on morphine tolerance by activation of kappa-opioid receptor (KOR). Preclinical studies also demonstrated that KOR agonists functionally attenuate addictive behaviors of morphine, such as conditioned place preference (CPP). Therefore, we hypothesize that PAT may inhibit morphine-induced CPP in rats.

Materials and methods

(1) Five groups of rats (n = 8 for each group) were alternately subcutaneous (s.c.) injected with morphine 10 mg/kg (one group receive normal saline as a control) and normal saline for 8 days and oral co-administrated with distilled water or PAT 0.3, 1.0, or 3.0 g/kg daily on days 2-9 during CPP training, respectively. (2) Other four groups of rats were randomly s.c. injected with nor-binaltorphimine (nor-BNI; 5 mg/kg) or normal saline (as a control) 120 min before alternately s.c. with morphine and normal saline and oral co-administrated with distilled water or PAT 3.0 g/kg daily. Each rat was acquired pre-conditioning and post-conditioning CPP data and assayed dynorphin concentrations by radioimmunoassay in rat's nucleus accumbens (NAc) after CPP training.

Results

(1) PAT 1.0 or 3.0 g/kg dose-dependently decreased the morphine-induced increase of CPP scores. (2) Nor-BNI completely antagonized the inhibition of PAT on morphine-induced CPP. (3) PAT dose-dependently increased dynorphin content in rats’ NAc after CPP training.

Conclusions

(1) PAT dose-dependently inhibited morphine-induced CPP. (2) The inhibition of PAT on morphine-induced CPP was probably due to activation of KOR by increasing dynorphin release in rats’ NAc.     

Anti-inflammatory activity of Trichosanthes cucumerina Linn. in rats

Aim of the study

Trichosanthes cucumerina Linn. (Family: Cucurbitaceae) is one of the medicinal plants that is often used in Sri Lankan traditional systems of medicine. One of its uses is the treatment of inflammatory conditions. However, validity of the anti-inflammatory activity has not been scientifically investigated so far. Therefore, the aim of this study was to investigate the anti-inflammatory potential of Trichosanthes cucumerina hot water extract (HWE) and its fractions.

Materials and methods

The anti-inflammatory activity of Trichosanthes cucumerina was evaluated by use of the carrageenan-induced paw oedema model in Wistar rats. In addition, the mechanism/s by which Trichosanthes cucumerina is mediated the anti-inflammatory activity was assessed by determining its effects on (a) membrane stabilizing activity and (b) nitric oxide inhibitory activity.

Results

Apart from the lowest dose of the HWE, other tested doses (500, 750, 1000 mg/kg) produced a significant (P ≤ 0.05) inhibition of the inflammation, most pronounced at 5 h after the injection of carrageenan. The anti-inflammatory effect induced by 750 mg/kg, was comparable to that of the reference drug, indomethacin at 4 and 5 h. Inhibition of nitric oxide (NO) production and membrane stabilization activities are probable mechanisms by which Trichosanthes cucumerina mediates its anti-inflammatory actions. Among the tested fractions, methanol fraction (MEF) and aqueous fraction (AQF) at a dose of 75 mg/kg exhibited marked inhibition against carrageenan-induced hind paw oedema. The anti-inflammatory effect induced by MEF, was comparable to that of the reference drug, indomethacin and as well as to the 750 mg/kg of HWE at 4 and 5 h.

Conclusions

(a) These findings rationalize the traditional usage of this plant as an anti-inflammatory agent and (b) membrane stabilizing properties and NO inhibitory activity are possible mechanisms through which Trichosanthes cucumerina mediates its anti-inflammatory action.     

Paeoniflorin inhibits function of synoviocytes pretreated by rIL-1α and regulates EP4 receptor expression

Ethnopharmacological relevance

To investigate the effect of the Paeoniflorin (Pae), a main active component of total glucosides of paeony (TGP) extracted from the root of Paeonia lactiflora, on regulation of synoviocytes cultured from rats collagen-induced arthritis (CIA) in vitro.

Materials and methods

CIA was induced in male Sprague-Dawley rats immunized with chicken type II collagen (CCII) in Freund's complete adjuvant. The levels of interleukin-1 (IL-1), tumor necrosis factor α (TNF-α), prostaglandin E₂ (PGE₂) and cyclic adenosine monophosphate (cAMP) were measured by radioimmunoassay. The proliferation responses was determined by the 3-(4,5-2dimethylthiazal-2yl) 2,5-diphenyltetrazoliumbromide (MTT) assay. Expression of E-prostanoid (EP₄) receptor was detected by Western blotting technique.

Results

Treatment of Pae (2.5, 12.5, 62.5 μg/ml) significantly decreased the production of IL-1 and TNF-α. Recombinant interleukin-1 (rIL-1α) (10 ng/ml) apparently stimulated synoviocyte, thymocyte and splenocyte proliferation, and Pae (12.5, 62.5 μg/ml) inhibited abnormal proliferation responses stimulated by rIL-1α. Moreover, rIL-1α time- and concentration-dependently increased production of PGE₂. The production of PGE₂ produced by synoviocytes from CIA rats significantly inhibited by administration of Pae (12.5, 62.5 μg/ml). rIL-1α (10 ng/ml) decreased cAMP of synoviocytes cells treated for 24 h. Similarly rIL-1α (0.1, 1, 10 ng/ml) induced a concentration-dependent decrease in the production of cAMP at 24 h. Pae (12.5, 62.5 μg/ml) increased the production of cAMP in synoviocytes. The immunoblot, Pae (12.5, 62.5 μg/ml) apparently increased the expression of EP₄ receptor in synoviocytes stimulated by rIL-1α (10 ng/ml).

Conclusions

The present study indicates that Pae might exert its anti-inflammatory effects through suppressing synoviocytes function and regulating immune cells responses in CIA rats, which might be associated with its ability to up-regulate the E-prostanoid (EP₄) receptor protein expression and modulate intracellular cAMP level.     

Safety evaluation of tea (Camellia sinensis (L.) O. Kuntze) flower extract: assessment of mutagenicity, and acute and subchronic toxicity in rats

Ethnopharmacological relevance

Tea (Camellia sinensis (L.) O. Kuntze, Theaceae) flowers possess many physiological functions and have been used in traditional medicines for deodorization, skin care, cough suppressant and expectorant in China. However, there is a little information about its possible toxicity.

Aim of the study

The present investigation was carried out to evaluate the safety of tea flower extract by mutagenicity and acute and subchronic toxicity studies.

Materials and methods

Mutagenicity of tea flower extract was evaluated by the Ames test in Salmonella typhimurium strains TA97, TA98, TA100 and TA102 at concentrations of 0.008, 0.04, 0.2, 1.0, 5.0 mg/plate. In the acute toxicity study, Sprague-Dawley rats were administered a single dose of 12.0 g/kg of body weight by gavage, and were monitored for 14 days. In the subchronic toxicity study, tea flower extract was administered by gavage at doses of 1.0, 2.0 and 4.0 g/kg body weight daily for 13 weeks to Sprague-Dawley rats.

Results

In the Ames test, there was no mutagenic effect of tea flower extract (up to 5.0 mg/plate) towards four tested strains (TA97, TA98, TA100, TA102), with or without metabolic activation (S9). In the acute toxicity study, all animals gained weight and appeared active and normal, so the LD₅₀ value must be >12.0 g/kg body weight. In the subchronic toxicity study, no dose-related effects on survival, growth, hematology, blood chemistry, organ weights, or pathologic lesions were observed.

Conclusion

These results indicate that tea flower extract does not possess mutagenic potential, and that both acute and subchronic toxicity towards animals is very low. A no-observed adverse-effect level (NOAEL) for tea flower extract is 4.0 g/kg bw/day for rats under the conditions of this study.     

Antinociceptive effect of the essential oil of Zingiber zerumbet in mice: possible mechanisms

Ethnopharmacological relevance

Zingiber zerumbet (L.) Smith, a wild edible ginger species or locally known as “lempoyang”, commonly used in the Malays traditional medicine as an appetizer or to treat stomachache, toothache, muscle sprain and as a cure for swelling sores and cuts.

Aim

The present study was conducted to investigate the possible mechanism of actions underlying the systemic antinociception activity of the essential oil of Zingiber zerumbet (EOZZ) in chemical-induced nociception tests in mice.

Materials and methods

Acetic acid-induced abdominal constriction, capsaicin-, glutamate- and phorbol 12-myristate 13-acetate-induced paw licking tests in mice were employed in the study. In all experiments, EOZZ was administered systemically at the doses of 50, 100, 200 and 300 mg/kg.

Results

It was shown that EOZZ given to mice via intraperitoneal and oral routes at 50, 100, 200 and 300 mg/kg produced significant dose dependent antinociception when assessed using acetic acid-induced abdominal writing test with calculated mean ID₅₀ values of 88.84 mg/kg (80.88-97.57 mg/kg) and 118.8 mg/kg (102.5-137.8 mg/kg), respectively. Likewise, intraperitoneal administration of EOZZ at similar doses produced significant dose dependent inhibition of neurogenic pain induced by intraplantar injection of capsaicin (1.6 μg/paw), glutamate (10 μmol/paw) and phorbol 12-myristate 13-acetate (1.6 μg/paw) with calculated mean ID₅₀ of 128.8 mg/kg (118.6-139.9 mg/kg), 124.8 mg/kg (111.4-139.7 mg/kg) and 40.29 (35.39-45.86) mg/kg, respectively. It was also demonstrated that pretreatment with l-arginine (100 mg/kg, i.p.), a nitric oxide precursor significantly reversed antinociception produced by EOZZ suggesting the involvement of l-arginine/nitric oxide pathway. In addition, methylene blue (20 mg/kg, i.p.) significantly enhanced antinociception produced by EOZZ. Administration of glibenclamide (10 mg/kg, i.p.), an ATP-sensitive K⁺ channel antagonist significantly reversed antinociceptive activity induced by EOZZ.

Conclusion

Together, the present results suggested that EOZZ-induced antinociceptive activity was possibly related to its ability to inhibit glutamatergic system, TRPV1 receptors as well as through activation of l-arginine/nitric oxide/cGMP/protein kinase C/ATP-sensitive K⁺ channel pathway.     

Bacopa monniera leaf extract up-regulates tryptophan hydroxylase (TPH2) and serotonin transporter (SERT) expression: implications in memory formation

Aim of the study

To examine the effect of Bacopa monniera leaf ethanolic extract (BMEE) on the serotonergic system of postnatal rats with reference to learning and memory.

Materials and methods

From postnatal day (PND)-15-29, rats were treated with BMEE (40 mg/kg BW + 0.5% gum acacia) by oral gavage. Behavioural tests (Y-maze, hole-board and passive avoidance) were used to evaluate their learning (PND-32-37) and retention of memory (PND-47-53). Effect of BMEE on neurotransmitter system was analyzed by ELISA and semi-quantitative polymerase chain reaction (PCR).

Results

Oral administration of BMEE improved learning and retention of memory significantly in all behavioural tasks. Following BMEE treatment, the level of serotonin (5-HT) increased while dopamine (DA) decreased significantly. We also found variation in the level of acetylcholine (ACh). However, no significant changes were observed in the level of ACh and glutamate (Glu). The level of 5-HT was significantly elevated up to PND-37 and was then restored to normal level on PND-53. Interestingly, concomitant up-regulation was recorded in the mRNA expression of serotonin synthesizing enzyme tryptophan hydroxylase-2 (TPH2) and serotonin transporter (SERT) on PND-29 and PND-37, which was restored on PND-53.

Conclusions

The results suggest that BMEE treatment significantly enhances the learning and retention of memory in postnatal rats possibly through regulating the expression of TPH2, 5-HT metabolism and transport.     

Anti-inflammatory activity of Eupatorium perfoliatum L. extracts, eupafolin, and dimeric guaianolide via iNOS inhibitory activity and modulation of inflammation-related cytokines and chemokines

Ethnopharmacological relevance

Eupatorium perfoliatum L. has been used traditionally for the treatment of fever, malaria and inflammation-associated diseases. Nowadays it is mostly used as immune activating remedy. The following study was performed to evaluate extracts with different polarity and defined lead-compounds from the herbal material on potential in vitro activities concerning immune cell activation, phagocytosis, and inflammation-related processes.

Materials and methods

MeOH-, EtOH-, and DCM extracts, beside several subfractions and isolated polysaccharides, sesquiterpene lactones and flavonoids were prepared and characterized analytically from the aerial parts of E. perfoliatum. Immunological activity was tested within lymphocyte transformation test on PBMC, test on enhancement of phagocytosis and of NO-production by murine RAW 264.7 macrophages. Anti-inflammatory effects were assessed from LPS-stimulated RAW 264.7 cells by NO/iNOS quantification, gene array, real-time PCR and ELISA.

Results

No stimulatory activity was found within lymphocyte transformation test, for phagocytic activity and NO formation in macrophages. MeOH-, EtOH- and DCM extracts showed anti-inflammatory activity against LPS-stimulated macrophages by inhibition of NO release (IC₅₀ > 100, 89, 19 μg/mL resp.) with eupafolin and a dimeric guaianolide having prominent NO inhibiting activity (IC₅₀ 6 resp. 16 μM). Anti-inflammatory activity was found on gene and protein level by significant down-regulation of cytokines CSF-3, IL-1α, IL-1β, and chemokines CCL2, CCL22 and CXCL10. Also TNF was down-regulated moderately (−17%).

Conclusions

Although the postulated immunostimulating properties of E. perfoliatum have not been confirmed, the anti-inflammatory effects can be seen as a verification of the traditional use against inflammatory diseases.     

Anti-ulcerogenic properties of Quassia amara L. (Simaroubaceae) standardized extracts in rodent models

Aim of the study

Quassia amara L. is commonly used in Costarican folk medicine. It has been used for the treatment of a broad range of gastrointestinal symptoms such as dyspepsia, gastritis and constipation. In this study, the gastroprotective activity of two standardized extracts of Quassia amara L., Lipro^® and Ligas^®, was evaluated.

Materials and methods

Anti-ulcerogenic properties were evaluated in female rats under acute ulcer-induction models (ethanol, indomethacin and hypotermic restraint). To get a deeper insight in the anti-ulcerogenic properties of the extracts, Ligas^® was evaluated in the Shay rat model. Five parameters were estimated with this model: gastric mucus barrier, non-protein sulfhydril groups (NPSG) in the gastric mucosa, and pH, total acidity and peptic activity of the gastric juice.

Results

Induction of ulcers by 95% ethanol (0.5 mL per os), indomethacin (100 mg/kg s.c.) and stress (2 h in hypothermic restraint) was inhibited significantly with administration of Lipro^® (p < 0.05), in a dosage range from 4.9 mg/kg/d to 48.9 mg/kg/d. Treatment was given for one week. The extract Ligas^® showed a significant augmentation of NPSG (p < 0.05) in a dosage range from 4.0 to 39.7 mg/kg. Ligas^® did not produce a significant change (p > 0.05) in the other indicators.

Conclusions

Quassia amara L. standardized extracts, Lipro^® and Ligas^®, showed an important anti-ulcerogenic effect in acute ulcer induction models. Their effect was related to an increase in gastric barrier mucus and non-protein sulfhydril groups.