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.     

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.     

Phaleria macrocarpa (Scheff.) Boerl fruit aqueous extract enhances LDL receptor and PCSK9 expression in vivo and in vitro

Ethnopharmacological relevance

Phaleria macrocarpa (Scheff.) Boerl (Pm) has been shown to reduce cholesterol level in vitro and in vivo experiment.

Aim of the study

This study investigated the effects of Pm fruit on weight control and mechanistic basis of its anti-hypercholesterolemic effect in both in vivo and in vitro.

Materials and method

In the in vivo study, thirty six male Sprague Dawley were randomized to six groups. Five groups were induced into hypercholesterolemia by giving 3% cholesterol enriched-diet for 52 days while one group acted as control. The rats were then treated with Pm extract (0, 20, 30 and 40 mg/ml) or simvastatin for 84 days. The following parameters were determined: (1) body weight, (2) blood lipid profile (total cholesterol, triglyceride, HDL and LDL) and (3) hepatic LDL receptor (160 kDa and 120 kDa) and PCSK9 proteins. In the in vitro study, HepG2 cells were cultured in serum-free RPMI supplemented with 0.2% BSA with or without LDL and in the presence of Pm extract (0, 0.1, 2, 40 and 1000 μg/ml) or simvastatin (4.60 μg/ml) for 24 h. The abundance of both LDL receptor and PCSK9 proteins and mRNA were investigated.

Results

Pm extract significantly (P < 0.05) reduced body weight gain, total cholesterol, triglycerides, HDL LDL levels and upregulated hepatic LDL receptor as well as PCSK9 proteins of hypercholesterolemic rats. These results were supported by studies in HepG2 cells whereby Pm extract also significantly upregulated both LDL receptor and PCSK9 at protein and mRNA levels.

Conclusion

This study enhances the potential usage of Pm fruit for controlling the body weight of obese people and for treating hypercholesterolemia.     

Hypoglycemic and antihyperglycemic effect of Witheringia solanacea in normal and alloxan-induced hyperglycemic rats

Aim of the study

Witheringia solanacea is a small shrub that belongs to the Solanaceae family. The plant is used as an antidiabetic in Costa Rican herbal medicine. The aim of this study was to evaluate the hypoglycemic and antihyperglycemic activity of the aqueous extract of W. solanacea leaves in rodent models.

Materials and methods

A crude extract of W. solanacea leaves was prepared in boiling water and the aqueous filtrate was lyophilized. A single oral dose of 250, 500 and 1000 mg/kg of the extract was evaluated for hypoglycemic activity in a glucose tolerance test in normal rats and for antihyperglycemic activity in alloxan-induced (140 mg/kg) diabetic rats. The blood glucose level was determined at different times by the glucose oxidase method.

Results

Dosage of 500 and 1000 mg/kg of the extract significantly decreased (p < 0.05) blood glucose levels in the glucose tolerance test in normal rats after 1 h, there was no significant difference observed at 250 mg/kg. Dose of 500 mg/kg of the extract significantly reduced (p < 0.05) blood glucose levels in alloxan induced hyperglycemic rats at 4 and 5 h.

Conclusions

In the present study, the hypoglycemic and antihyperglycemic potential of the W. solanacea was demonstrated in rats. These results give support to the traditional use of W. solanacea as antidiabetic herbal medicine.     

The effects of Sceletium tortuosum in an in vivo model of psychological stress

Aim of the study

Sceletium, and especially Sceletium tortuosum, is traditionally used as masticator and thought to have a sedative effect which may be beneficial to reduce symptoms of anxiety and/or depression. The current study evaluated the scientific merit of these anecdotal claims in an in vivo model of psychological stress.

Materials and methods

Male Wistar rats were administered either placebo, 5 or 20 mg/kg/day of Sceletium tortuosum extract for 17 days by daily oral gavage. 50% of rats were exposed to repeated restraint stress lasting 1 h for the last 3 days of treatment. Rat behavioral changes in response to stress were assessed using the elevated plus maze on the last day of restraint, immediately after the restraint session. Rats were sacrificed 24 h after the last restraint exposure and whole blood collected.

Results

Behavior indicated a limited effect of lower dose Sceletium to decrease restraint stress-induced self-soothing behavior, as well as to decrease stress-induced corticosterone levels. However, increased IL-1β levels argue against the claim that the plant may act as selective serotonin reuptake inhibitor, while this result combined with increased levels of C-reactive protein and prostaglandin E₂ suggest intolerance to the treatment. Decreased IL-2 and increased IL-10 levels in response to Sceletium treatment suggest a suppressive effect on T helper 1 immune function.

Conclusions

Although data indicates a limited positive effect of Sceletium on restraint-induced anxiety, numerous side-effects were evident. More research is required to derive an optimal therapeutic dose.     

Anti-hyperglycemic activity of the leaves of Tetracera scandens Linn. Merr. (Dilleniaceae) in alloxan induced diabetic rats

Aim of the study

The present study was aimed to investigate the anti-diabetic potential of the leaves of Tetracera scandens Linn. Merr. (Dilleniaceae) in vivo with regard to prove its efficacy by local herbalists in the treatment of diabetes frailties.

Materials and methods

Crude aqueous (AQ) and methanol (MEOH) extracts of the leaves of T. scandens L. were administered to both normal and alloxan induced diabetic male albino rats (Wistar strain). The blood glucose levels were measured at 0, 2, 4, 6 and 8 h after oral administration of AQ and MEOH extracts.

Results

Significant reduction in glucose was observed in fasting blood glucose levels in the treated diabetic rats without causing any hypoglycemic effect compared to normal rats. Both polar extracts of the leaves of T. scandens L. exhibited significant anti-hyperglycemic activity at different doses and intervals. The highest anti-hyperglycemic effect (62.5%) was observed by the AQ extract at 0.25 g/kg body weight (b.w.) and MEOH extract (36.5%) at 0.5 g/kg b.w. after 8 h. The significant anti-hyperglycemic activity was found to be comparable with a known oral synthetic hypoglycemic drug, glibenclamide 0.25 mg/kg b.w.

Conclusion

This study provides scientific evidence that the leaves of T. scandens L. have anti-diabetic efficacy and justifies its utility by local herbalists. However, more experiments at the clinical levels are required to confirm the utility of this plant by traditional practitioners in the management of diabetes mellitus.     

The analgesic and anti-inflammatory effect of WIN-34B, a new herbal formula for osteoarthritis composed of Lonicera japonica Thunb and Anemarrhena asphodeloides BUNGE in vivo

Ethnopharmacological relevance

: Lonicera japonica Thunb and Anemarrhena asphodeloides BUNGE have been used for the treatment of a variety of inflammatory diseases, cold and infective diseases in many countries, including Korea and China.

Aim of the study

: This study aimed to assess the anti-nociceptive and anti-inflammatory activities of n-butanol fraction (WIN-34B) prepared from dried flowers of Lonicera japonica and dried roots of Anemarrhena asphodeloides as potential novel treatment of osteoarthritis.

Materials and methods

: Anti-nociceptive activities of WIN-34B (100, 200 and 400 mg/kg, p.o.) were measured using acetic acid-induced writhing response, formalin-induced paw licking, hot plate, radiant heat tail-flick, carrageenan-induced paw pressure, and Hargreaves tests, respectively. Anti-inflammatory activities of WIN-34B (100, 200 and 400 mg/kg, p.o.) were assessed using acetic acid-induced vascular permeability, carrageenan-induced paw edema, and croton oil-induced ear edema. Anti-osteoarthritis effect of WIN-34B was analyzed using monosodium iodoacetate (MIA)-induced osteoarthritis animal model.

Results

: WIN-34B exhibited better anti-inflammatory activity than that of celecoxib in carrageenan at the dose of 200 mg/kg and croton oil-induced paw edema and ear edema at the doses of 200 and 400 mg/kg. WIN-34B exhibited significant anti-inflammatory effects on vascular permeability. WIN-34B also exhibited significant anti-nociceptive activities in the late phase of formalin-induced paw licking and writhing response model in mice. In radiant heat tail-flick and carrageenan-induced paw pressure tests, WIN-34B at the dose of 400 mg/kg and at the doses of 200 and 400 mg/kg presented similar activities to indomethacin and celecoxib. Compared to indomethacin WIN-34B at 400 mg/kg showed similar or better anti-nociceptive activities after 1 and 2 h of theraphy in the hot plate test and better anti-nociceptive activity than that of celecoxib in Hargreves test. In the MIA-induced osteoarthritis animal models, WIN-34B at 400 mg/kg exhibited similar or better anti-nociceptive property than that of celecoxib throughout the pain measurement periods.

Conclusion

: When compared to celecoxib, WIN-34B exhibited similar or better anti-nociceptive and anti-inflammatory activities in osteoarthritic animal models, which may become a potential novel treatment for osteoarthritis.     

Analgesic effects and anti-inflammatory properties of the crude methanolic extract of Schwenckia americana Linn (Solanaceae)

Aim of the study

To evaluate analgesic effect and anti-inflammatory properties of Schwenckia americana (Solanaceae), a medicinal plant used for treating rheumatic pains and swelling in North-western Nigeria.

Materials and methods

Three doses (25 mg/kg, 50 mg/kg and 100 mg/kg) of the crude methanolic extract of Schwenkia americana were evaluated for analgesic and anti-inflammatory activities using acetic acid induced writhing test, formalin induced nociception, and formalin induced hind paw oedema in rats.

Results

All doses (25, 50, 100 mg/kg) of the extract tested were effective. The extract at the tested doses produced a percentage inhibition of the acetic acid induced abdominal constriction of (53.3, 58.0 and 86.7%), respectively. A percentage inhibition of the formalin induced nociception of 44.00, 56.04, and 56.04% (early phase) and 33.00, 36.63 and 59.71% (late phase) was also produced. The inhibition of oedema formation increased with increasing dosage from 25 to 100 mg/kg. The crude extract produced a statistically significant analgesic and anti-inflammatory activity comparable to the effect of standard drug (10 mg/kg Piroxicam).

Conclusion

This study demonstrated the potential analgesic and anti-inflammatory properties of crude methanolic extract of Schwenkia americana thus justifying its traditional usage.     

Antidepressant-like activity of the aqueous extract of Allium macrostemon in mice

Aim of the study

The aim of this study was to identify the effects of water extracts of Allium macrostemon Bunge (AM-W), a traditional herb, in mice.

Materials and methods

The antidepressant-like activities of AM-W were evaluated through behavioral despair in forced swimming test and tail suspension test. To elucidate the mode of action of the antidepressant-like effects of AM-W, new born cells in the subgranular zone and the granule cell layer were analyzed by immunostaining for incorporation of 5-bromo-2-deoxyuridine (BrdU). In addition, the effects of AM-W on the expression levels of brain-derived neurotrophic factor (BDNF) were investigated by western blotting and immunohistochemistry.

Results

The administration of AM-W reduced the immobility duration in the forced swimming test and tail suspension test (100 or 200 mg/kg, P < 0.05). Sub-chronic administration of AM-W (100 or 200 mg/kg, p.o., for 14 days) increased the number of BrdU-incorporating cells. The percentage of BrdU-incorporating cells co-localized with NeuN was significantly increased after AM-W administration (100 or 200 mg/kg, P < 0.05). Moreover, the expression levels of BDNF which is reported to be associated with neurogenesis were significantly increased in the hippocampus after administration of AM-W.

Conclusions

These results suggest that AM-W may be a good antidepressant, and that its mechanism of action may be related to its positive effects on neurogenesis and BDNF release.     

Neuroprotective effects of water-soluble Ganoderma lucidum polysaccharides on cerebral ischemic injury in rats

Aim of the study

To investigate the neuroprotective effects of water-soluble Ganoderma lucidum polysaccharides (GLPS) on cerebral ischemic injury in rats, and to explore the involved mechanisms.

Materials and methods

Two models [middle cerebral artery occlusion (MCAO) in Sprague-Dawley (SD) rats and oxygen and glucose deprivation (OGD) in primary cultured rat cortical neurons] were employed to mimic ischemia-reperfusion (I/R) damage, in vivo and in vitro, respectively. Cerebral infarct area was measured by tetrazolium staining, and neurological functional deficits were assessed at 24 h after I/R. Neuronal apoptosis was studied by Nissl staining and DNA fragmentation assay. Neuronal injury was assessed by morphological examination using phase-contrast microscopy and quantified by measuring the amount of lactate dehydrogenase (LDH) leakage, cell viability was measured by sodium 3′-1- (phenylaminocarbonyl)-3, 4-tetrazolium-bis (4-methoxy-6-nitro) benzene sulfonic acid (XTT) reduction. Neuronal apoptosis was determined by flow cytometry, and electron microscopy was used to study morphological changes of neurons. Caspase-3, -8 and -9 activation and Bcl-2, Bax protein expression were determined by western blot analysis.

Results

Oral administration of GLPS (100, 200 and 400 mg/kg) significantly reduced cerebral infarct area, attenuated neurological functional deficits, and reduced neuronal apoptosis in ischemic cortex. In OGD model, GLSP (0.1, 1 and 10 μg/ml) effectively reduced neuronal cell death and relieved cell injury. Moreover, GLPS decreased the percentage of apoptotic neurons, relieved neuronal morphological damage, suppressed overexpression of active caspases-3, -8 and -9 and Bax, and inhibited the reduction of Bcl-2 expression.

Conclusions

Our findings indicate that GLPS protects against cerebral ischemic injury by inhibiting apoptosis by downregulating caspase-3 activation and modulating the Bcl-2/Bax ratio.     

In vitro antimicrobial, anthelmintic and cyclooxygenase-inhibitory activities and phytochemical analysis of Leucosidea sericea

Ethnopharmacological relevance

Leucosidea sericea is used as a vermifuge and in the treatment of ophthalmia by various tribes in southern African countries.

Aim of the study

The study aimed at screening leaves and stems of Leucosidea sericea for pharmacological activity and validating the plant's traditional use. A general phytochemical screening was also carried out.

Materials and methods

Petroleum ether (PE), dichloromethane (DCM), ethanol (EtOH) and water extracts of the plant parts were investigated for antimicrobial, anthelmintic and cyclooxygenase (COX) inhibitory activities. Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae) and Candida albicans were used for the antimicrobial evaluation. Caenorhabditis elegans was used for the anthelmintic assay using the microdilution technique. Cyclooxygenase-1 and -2 (COX-1 and -2) were used to evaluate the anti-inflammatory potential of the plant extracts. Phytochemical analysis for phenolic compounds, including gallotannins, condensed tannins and flavonoids was done using 50% methanol extracts of the leaves and stems employing spectrophotometric methods.

Results

The leaf extracts exhibited broad spectrum antibacterial activity ranging from 0.025 to 6.25 mg/ml. The most noteworthy minimum inhibitory concentration (MIC) of 0.025 mg/ml was exhibited by PE and DCM leaf extracts against Bacillus subtilis and Staphylococcus aureus, respectively. In the anthelmintic assay, the best minimum lethal concentration (MLC) value of 0.26 mg/ml was observed for the DCM and EtOH leaf extracts. Both leaf and stem organic solvent extracts exhibited high to moderate inhibition against COX-1 and -2 at a screening concentration of 250 μg/ml. At lower concentrations, the extracts displayed a dose-dependent inhibition, with the lowest IC₅₀ values of 0.06 μg/ml (COX-1) and 12.66 μg/ml (COX-2) exhibited by the PE extract of the leaves. Generally, the leaf extracts exhibited better pharmacological activities and contained higher amounts of phenolic compounds than the stem extracts. Alkaloids and saponins were only detected in the leaf and stem extracts, respectively.

Conclusion

The reported results support the local use of Leucosidea sericea against eye infections and as a vermifuge. The pharmacological activities exhibited by the leaf extracts are probably due to their higher phenolic levels.     

Evaluation of safety of the herbal formula Ojeok-san: Acute and sub-chronic toxicity studies in rats

Aim of the study

Ojeok-san (OJS; wuji powder in China and goshaku-san in Japan), a widely used herbal formula in traditional Korean medicine and Japanese herbal medicine (Kampo medicine), has been used to treat common cold and illnesses including fatigue and gastrointestinal disorders, but there is very little information on its safety. To provide information on the safety of OJS, we evaluated its acute and sub-chronic toxicity in rats.

Materials and methods

The single and sub-chronic toxicity of OJS was examined using male and female Sprague-Dawley rats. The rats were treated with the OJS extract orally at the highest dose level of 2000 mg/kg/day body weight. After single administration, signs of toxicity were observed every hour for the first 6 h and every day for two weeks. In the sub-chronic toxicity study, OJS was administered for 13 weeks. Mortality, clinical signs, body weight changes, food and water consumption, ophthalmologic findings, urinalysis, hematological and biochemical parameters, gross findings, organ weights and histological markers were monitored during the study period.

Results

We found no mortality and no abnormality in clinical signs, body weight, and necropsy findings for any of the animals in the acute and sub-chronic toxicity study following oral administration of OJS.

Conclusion

OJS may not have any single dose toxicity. The lethal dose with a 50% mortality rate (LD₅₀) was over 2000 mg/kg. The no-observed adverse effects level (NOAEL) was considered to be 2000 and 1000 mg/kg/day for male and female rats, respectively.     

Hypoglycemic effect of a leaf extract of Pseuderanthemum palatiferum (Nees) Radlk. in normal and streptozotocin-induced diabetic rats

Ethnopharmacological relevance

Pseuderanthemum palatiferum (Nees) Radlk (Acanthaceae) was first found in Northern Vietnam and expanded throughout the country including the Mekong Delta region. The leaves of this plant are recommended in folk medicine of Vietnam and Thailand for promoting and treating various diseases including hypertension, diarrhea, arthritis, hemorrhoids, stomachache, tumors, colitis, bleeding, wounds, constipation, flu, colon cancer, nephritis, and diabetes.

Aim of the study

The hypoglycemic effect of an 80% ethanolic leaf extract from the leaves of Pseuderanthemum palatiferum (PPE) was investigated in normal and streptozotocin (STZ)-induced diabetic rats.

Materials and methods

The PPE was administered daily and orally to the rats at the doses of 250, 500, and 1000 mg/kg body weight (b.w.) for 14 days. The levels of fasting plasma glucose (FPG), serum insulin, and biochemical data such as blood urea nitrogen (BUN), triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and alkaline phosphatase (ALP) were evaluated. The hypoglycemic effect of PPE was compared to that of the known anti-diabetic drug glibenclamide (0.25 mg/kg b.w.).

Results

FPG and serum insulin in normal rats were not significantly different from the control and test groups in all dosages. The treated diabetic rats which had received PPE and glibenclamide showed significantly (p < 0.05) decreased FPG and increased serum insulin levels at the end of the experiment. The hypoglycemic effect of PPE at the dose of 250 mg/kg b.w. was significantly (p < 0.05) more effective than that of glibenclamide. The serum insulin in PPE fed diabetic rats at the dose of 250 mg/kg b.w. was not different from those which had received glibenclamide, and this dose was significantly (p < 0.05) more effective than PPE at the doses of 500 and 1000 mg/kg b.w. while PPE increased HDL and decreased TC, TG, LDL, BUN and ALP in the diabetic rats.

Conclusions

PPE has a beneficial effect in hyperglycemic rats and may prevent the complication of diabetes.     

Antinociceptive and gastroprotective actions of ethanolic extract from Pluchea sagittalis (Lam.) Cabrera

Ethnopharmacological relevance

Pluchea sagittalis, an herbaceous plant widely distributed in South America, is used in folk medicine for the treatment of digestive diseases and inflammation.

Aim of the study

This study was designed to investigate the antinociceptive and gastroprotective effects of the ethanolic extract (EE) of aerial parts from Pluchea sagittalis in rodents.

Materials and methods

The antinociceptive effects of EE was evaluated in mice after oral administration in chemical tests (acetic-acid, glutamate and formalin) or by biting behavior following intrathecal administration of cytokines such as interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) in mice. Furthermore, rats were treated with EE and subsequently exposed to acute gastric lesions induced by 80% ethanol. Afterwards the gastric lesion extension and the mucus levels of gastric mucosa were measured.

Results

The oral administration of EE showed a dose-dependent inhibition of acetic acid-induced abdominal constrictions and glutamate-induced pain in mice, with ID₅₀ values of 624.0 (523.0-746.0) mg/kg and 368.0 (216.0-628.0) mg/kg, respectively. In the formalin test, the EE also produced significant inhibition of the inflammatory phase, with an ID₅₀ value of 411.0 (183.0-721.0) mg/kg; however, it was ineffective in the neurogenic phase caused by formalin. In addition, oral treatment with EE caused a significant inhibition of biting behavior induced by i.t. injection of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). The antinociception caused by the EE (300 mg/kg, p.o.) was not reversed by naloxone (1 mg/kg, i.p.) when assessed in the acetic acid writhing test. The EE (300-1000 mg/kg, p.o.) did not affect the motor coordination of animals in an open-field model. Oral treatment with the EE protected rats against gastric lesions induced by ethanol, with an ID₅₀ value of 55.0 (46.6-64.9) mg/kg, and increased the mucus levels of gastric mucosa to levels found in the non-lesioned group.

Conclusions

The mechanism by which the extract produced antinociception still remains unclear, but this effect seems to be primarily related to the modulation or inhibition of the action of pro-inflammatory mediators. Furthermore, these data support, at least in part, the ethnomedical use of Pluchea sagittalis.     

Typhonium flagelliforme induces apoptosis in CEMss cells via activation of caspase-9, PARP cleavage and cytochrome c release: Its activation coupled with G0/G1 phase cell cycle arrest

Ethnopharmacological relevance

The plant Typhonium flagelliforme (TF), commonly known as ‘rodent tuber’ in Malaysia, is often used as traditional remedy for cancer, including leukemia.

Aim of the study

We had previously identified morphologically that the linoleic acid rich fraction (DCM/F7) from the tubers of this plant induces selective anti-proliferative effects and apoptosis in CEMss cells. In this present study, we subjected the same DCM/F7 fraction to cell based activity analyses in order to determine the possible mechanism of cell death in leukemic CEMss cells in vitro.

Materials and methods

Extraction of Typhonium flagelliforme tuber has done and fractionation has been done by vacuum liquid column chromatography. The anti-proliferative activity was assayed using MTT and the apoptosis detection was done by Annexin V and DNA laddering assay. Colorimetric caspase assay and immunoblot analysis were employed to detect the expression of protein associated with cell death. Cell cycle analysis was done using flow cytometry.

Results

We found that the cancer inhibitory effect of the DCM/F7 fraction in CEMss cells was 3 ± 0.08 μg/ml (IC₅₀). An early apoptotic induction in CEMss cells was observed by Annexin V assay, which showed a clear dose-dependent DNA fragmentation being observed in gel electrophoresis at 10 and 20 μg/ml. The DCM/F7 fraction at 3 μg/ml significantly arrested CEMss cells at G0/G1 phase (p < 0.05). A constant but increasing pattern-related Sub-G0/G1 index was observed between 12 and 72 h treatment. In relation to this, we further investigated the biochemical events leading to cell death and found that the DCM/F7 fraction increased the cellular levels of caspase-3 and -9 on treated cells. Our results indicated that cytochrome c from mitochondria into the cytosol increased gradually as the DCM/F7 concentration increases, which later lead to the subsequent cleavage of PARP in to 85 kDa fragments. On the contrary, Bcl-2 protein was found to decrease concomitantly during treatment.

Conclusions

Collectively, results presented in this study demonstrated that the DCM/F7 fraction inhibited the proliferation of leukemia cells, leading to the programmed cell death, which was confirmed to be through the mitochondrial pathway.     

Antinociceptive activity of methanol extract of fruits of Capparis ovata in mice

Ethnopharmacological relevance

Capparis ovata Desf. and Capparis spinosa L. have wide natural distribution in Turkey and they are consumed in pickled form. Flower buds, root bark, and fruits of the plant are used in folk medicine due to their analgesic, wound healing, cell regeneration, tonic, and diuretic effects.

Aim of the study

In this study, we attempted to identify the possible antinociceptive action of methanol extract prepared from fruits of Capparis ovata.

Materials and methods

Using tail immersion, hot plate and writhing tests, the antinociceptive effect of the methanol extract of Capparis ovata (MEC) fruits was assessed after intraperitoneal administration into mice. Morphine sulfate (5 mg/kg; i.p.) and diclofenac (10 mg/kg; i.p.) were used as reference analgesic agents. Naloxone (5 mg/kg; i.p.) was also tested.

Results

MEC was studied at the doses of 50, 100, and 200 mg/kg (i.p.) and exhibited significant antinociceptive activities in all tests used. The above-mentioned doses of the extract reduced the writhing responses by 32.21, 55.70, and 68.36%, respectively. MPE% were increased by 7.27, 12.07, 14.60% in the tail immersion, and 7.88, 11.71, 16.73% in the hot plate test at the tested doses, respectively. Naloxone antagonized antinociceptive effect at the doses of 100 and 200 mg/kg whereas partially antagonized the effect of MEC at the dose of 50 mg/kg.

Conclusions

Based on the results obtained, it can be concluded that MEC has antinociceptive effects both at the peripheral and central levels.     

Suppression of dust mite extract and 2,4-dinitrochlorobenzene-induced atopic dermatitis by the water extract of Lindera obtusiloba

Ethnopharmacological relevance

The Lindera obtusiloba has been used in traditional medicine for the treatment of inflammation and dermatitis. In this study, we investigated the effect of topical application of Lindera obtusiloba water extract (LOWE) on the house dust mite extract (Dermatophagoides farinae extract, DFE) and 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis (AD).

Materials and methods

We established AD model in BALB/c mice by repeated local exposure of DFE/DNCB to the ears. After a topical application of LOWE on the skin lesions, the epidermal thickness, mast cell infiltration, and serum immunoglobulin E (IgE) and histamine were measured. In addition, the gene expression of interleukin (IL)-4, IL-13, IL-31, and tumor necrosis factor (TNF)-α in the ears was assayed.

Results

LOWE reduced AD symptoms based on ear thickness, histopathological analysis, and serum IgE levels. LOWE inhibited mast cell infiltration into the ear and elevation of serum histamine in AD model. Moreover, LOWE suppressed DFE/DNCB-induced expression of IL-4, IL-13, IL-31, and TNF-α in the ears.

Conclusions

Our results showed that topical application of LOWE exerts beneficial effects in AD symptoms, suggesting that LOWE might be a candidate for the treatment of AD.     

Antimalarial activity and toxicity evaluation of a quantified Nauclea pobeguinii extract

Aim of the study

To evaluate the in vitro and in vivo antiplasmodial activity and toxicity of the aqueous and 80% EtOH extract of the stem bark of Nauclea pobeguinii (Pob. Ex. Pell.) Petit (Rubiaceae), a plant used in traditional medicine in DR Congo against malaria.

Materials and methods

The aqueous and 80% EtOH extract from N. pobeguinii stem bark, and its constituents (5S)-5-carboxystrictosidine, 19-O-methylangustoline, 3-O-β-fucosylquinovic acid, 3-ketoquinovic acid and strictosamide, were evaluated for their in vitro activity against Plasmodium falciparum (chloroquine-sensitive Ghana-strain). The 80% EtOH extract, containing 5.6% strictosamide, was evaluated in vivo in the 4-day P. berghei mouse model, and in the P. yoelii N67 model.

Results

All compounds were inactive or only moderately active in vitro. The aqueous and 80% EtOH extract displayed moderate in vitro activity with IC₅₀ values of 44 and 32 μg/mL, respectively, without apparent cytotoxicity on MRC-5 cells (CC50 > 64 μg/mL). Daily oral dosing of the 80% EtOH extract, at 300 mg/kg, resulted in 86% reduction of parasitaemia in the 4-day P. berghei mouse model, and 75% reduction in the P. yoelii N67 model. Prolonging oral dosing to 2 × 5 days, with an interval of 2 days, and oral administration of the 80% EtOH extract at 300 mg/kg induced 92% reduction of parasitaemia, and a mean survival time of 17 days. Strictosamide, the putative active constituent, may be metabolically activated in the gastrointestinal tract after oral administration. Levels of creatinin, urea, ALAT and ASAT remained unchanged after treatment. No acute toxicity was observed in mice after a single 2 g/kg oral dose, nor after 4 weekly doses. No significant macroscopic or microscopic lesions were observed in heart, lung, spleen, kidney, liver, large intestine and brain.

Conclusions

These results can partly support and justify the use of N. pobeguinii in traditional medicine in the DR Congo for the treatment of uncomplicated malaria.     

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.