Assessment of acute, sub-acute, chronic and genotoxicity of polyherbal formulation DCD-684 in mice.

Digas colic drops (DCD-684) a polyherbal formulation containing Carum carvi, Foeniculum vulgare, Mentha arvensis, Mentha piperita and Zingiber officinale is widely used in Pakistan against gastrointestinal ailments including infantile colic. The DCD-684 (0.03-3ml/kg.bw) administered orally in acute (7-days) and sub-acute toxicity (14-days) tests, displayed neither mortality nor toxicological changes in physical, behavioral, biochemical and histopathological parameters. In chronic study (90-days), DCD-684 (0.3-12ml/kg.bw) also revealed no changes. However, at 18 and 36 ml/kg.bw, liver demonstrated mild inflammation correlating with raised aspartate transaminase (AST), alkaline phosphatase (ALP) and alpha fetoprotein (AFP) levels. Increased levels of urea and inflamed renal parenchyma indicated mild nephro-toxicity with high alanine aminotransferase (ALT) at 36ml/kg.bw. The LD50 of DCD-684 in mice was 27.5 ml/kg.bw. In hepatocytes at 36ml/kg.bw, elevated mRNA expression of pro-inflammatory chemokines and cytokines were evident. DCD-684 neither damaged DNA nor induced cytotoxicity in micronucleus assay. In conclusion, polyherbal DCD-684 caused neither hepatic, renal, genotoxicity nor any undesirable effect in mice. Higher doses administered for 90 days showed mild toxic effects with no sign of necrosis, fibrosis or genotoxicity. Thus, in mice DCD-684 demonstrated a wide margin of safety to be used for the relief of infantile colic.

Analysis of Unani coded formulation on the hormonal parameters of patients with polycystic ovarian syndrome.

The effect of Unani coded polyherbal formulations (Picolin) containing: 1) Cinnamomum verum (Bark), 2) Glycyrrhiza glabra (Root), 3) Linum usitatissimum (Seed) and 4) Vitex agnus castus (Seed) on the hormonal levels of polycystic ovarian syndrome (PCOS) patients (n=73) was determined. A randomized controlled multi-center trial was conducted on three groups that received either: 1) Capsule Picolin (n=25, 500 mg, two capsules thrice a day) and 2) Hydroalcoholic extract, tablet Picolin-E (n=23, 250 mg thrice a day) or 3) Metformin (n=25, 500 mg, twice a day) that continued for 12 weeks. The effect of capsule Picolin before and after treatment on serum insulin (14.8±1.8 vs 10.7±1.7µIU/mL) and prolactin (17.7±0.9 vs 8.5±1.1ng/mL) levels were significant. Likewise, the tablet Picolin-E also demonstrated significant effect on serum insulin (16.7±1.7 vs 9.3±0.1µIU/mL) and prolactin (18.5±1 vs 10±1.5ng/mL) levels. On the contrary in metformin treated group these parameters, serum insulin (17.4±1.6 vs 16.4±1.4µIU/mL) and prolactin (21.7±2.8 vs 21±2.8ng/mL) were non-significant. The improvement in the menstrual flow in the PCOS patients after treatment with either capsule Picolin, tablet Picolin-E or metformin were improved by 84%, 61% and 54%, respectively. Unani formulation were comparatively more effective than metformin. The treatment effectiveness was capsule Picolin> tablet Picolin-E>metformin.

Antispasmodic activity and mechanism of action of polyherbal formulation DCD-684 on rabbit jejunum.

Digas colic drops (DCD-684) is a polyherbal formulation containing decoctions of five medicinal plants namely Carum carvi L., Foeniculum vulgare Mill, Mentha arvensis L., Mentha piperita L. and Zingiber officinale Roscoe. These plants have been extensively used in traditional medicine for the treatment of various gastrointestinal diseases including abdominal colic. This study was conducted to determine the spasmolytic effect of DCD-684 (100% v/v) and its individual plant components on isolated rabbit jejunum (in vitro) and their possible mechanism of action. The effects were evaluated on spontaneous and pre-contracted tissues using KCl (80mM) and other contractile agonists including acetylcholine (0.3µM), carbamylcholine (0.3µM), serotonin (10 µM) and histamine (100µM) in the presence and absence of DCD-684. The various concentrations of DCD-684 (0.1-3% v/v) demonstrated spasmolytic effects on both spontaneous (IC50=0.75%) and KCl-induced contractions (IC50=1.6%), respectively. It also inhibited the contractions induced by acetylcholine (IC50=0.45%), carbamylcholine (IC50=0.95%), serotonin (IC50=0.95%) and histamine (IC50=0.87%). The DCD-684 exhibited synergistic effect due to its five plant components suggesting that spasmolytic cascade is probably governed by muscarinic and/or nicotinic receptors, serotonergic histaminergic, as well as calcium channel blocking mechanisms. Thereby, providing the pharmacological basis of its therapeutic use in the gastrointestinal motility disorders and related inflammatory ailments.

Toxicological assessment of Opuntia dillenii (Ker Gawl.) Haw. cladode methanol extract, fractions and its alpha pyrones: Opuntiol and opuntioside.

ETHNOPHARMACOLOGICAL RELEVANCE: The edible plant Opuntia dillenii (Ker Gawl.) Haw. commonly known as Nagphana, belongs to the Cactaceae family. It is traditionally used to treat various ailments including inflammation, gastric ulcers, diabetes, hepatitis, asthma, whooping cough and intestinal spasm. AIM OF THE STUDY: Despite its traditional use in various countries, detailed toxicological studies of O. dillenii cladode are few. Thus in the current study, toxicity of O. dillenii cladode derived methanol extract, fractions and its α-pyrones: opuntiol and opuntioside have been addressed. METHODS: The test agents were assessed using both in vitro and in vivo toxicity assays. MTT on human embryonic kidney cell line (HEK-293), tryphan blue exclusion in rat neutrophils, Cytokinesis-B block micronucleus (CBMN) in human lymphocytes and genomic DNA fragmentation using agarose gel electrophoresis were performed. In acute toxicity test, mice orally received extract (5 g/kg) for 7 days followed by measurements of relative organ weight, biochemical (blood profile, liver and kidney function test) and histological studies (liver and kidney) were carried out. Rat bone marrow micronucleus genotoxicity assay was also conducted. RESULTS: O. dillenii derived test agents were non-cytotoxic and had no effect on the integrity of DNA. Methanol extract (5 g/kg) orally administered in mice did not cause any significant change in relative organ weights, biochemical parameters and liver and kidney histology as compared to vehicle control. In parallel, extract did not stimulate micronuclei formation in rat bone marrow polychromatic erythrocytes. CONCLUSION: These results led to conclude that edible O. dillenii extract is non-toxic via the oral route and appears to be non-cyto-, hepato-, nephro- or genotoxic, thereby supporting its safe traditional use against various ailments. Therefore, opuntiol and opuntioside may serve as lead compounds in designing new drug(s) derived from edible plants.

Jiawei Shengmai San herbal formula ameliorates diabetic associate cognitive decline by modulating AKT and CREB in rats.

Memory loss is a complication of diabetes which requires new approaches to its treatment. Shengmai San (SMS) is a famous traditional Chinese formula containing Panax ginseng, Ophiopogon japonicas, and Schisandra chinensis, whereas Radix puerariae has many reported pharmacological uses. In this study the combination, as Jiawei SMS (J-SMS) was screened for its ability to reverse diabetes-associated cognitive decline in rats. This was assessed behaviorally in diabetic rats (Streptozotocin, 45 mg/kg), with biochemical and western blot analysis (Akt and CREB). Diabetic rats showed fasting blood glucose (FBG) in the range of 13-15 mM throughout the study. J-SMS (0.5, 1.5, 4.5 g/kg) treatment significantly improved learning and memory deficit among diabetic rats as evidenced by preference for novel object, reduced escape latency and increased number of platform crossings (p < .05) in the NORT and MWM tests. Treatment with J-SMS also significantly improved the histopathological changes in the diabetic brain and increased the protein expression of AKT and CREB, required for proper memory function (p < .01). This study highlighted that J-SMS can reverse reference and working memory deficit among diabetic rats by modulating AKT and CREB proteins activation. Thus, J-SMS formulation might be possible candidate for further development.

Volatile constituents and in vitro activity of Syzygium aromaticum flower buds (clove) against human cancer cell lines.

The methanolic extract (SA-EXT) of Syzygium aromaticum flower buds and its fractions tested against three human cancer cell lines viz uterine cervix (HeLa), breast (MCF-7) and lung NCI (H-460) using sulforhodamine-B assay. The ethyl acetate soluble sub fraction (SA-EAS) was active only against HeLa cells with GI50value of 36± 3.4µg/mL. The most active sub-fraction (SA-PES-Fr-2) showed growth inhibition (GI50: 36, 50 and 68µg/ml against MCF-7, HeLa and NCI-H-460 cancer cell lines, respectively) with cytotoxic effect LC50= 88 ± 3.4 µg/mL against HeLa and LC50=86 ± 2.8 µg/mL against MCF-7. The most active sub-fraction (SA-PES-Fr-2) analyzed by GC and GC-MS techniques revealed that eugenol was most abundant (85.34%) along with minor constituents. Thus it can be concluded that growth inhibitory and cytotoxic effect residing in Syzygium aromaticum flower buds (clove) is more likely due to eugenol.

Cytotoxic and genotoxic action of Tagetes patula flower methanol extract and patuletin using the Allium test.

Tagetes patula is used to treat cancer patients in alternative healthcare systems. However, its cytotoxic and genotoxic effects have not been reported. Therefore, themethanol extract of T. patula flower, the ethyl acetate fraction, and the pure compound patuletin were evaluatedusing the Allium test.The methanol extract and fraction contained ~3% and ~36% patuletin, respectively, with ~98% purity. The methanol extract caused inhibition of Allium root growth displaying an IC50 value of ~500 µg/mL, while the fraction and patuletin were more potent by ~2 and ~5 times, respectively. The Allium root tips demonstrated a decline in prophase, metaphase, anaphase, and telophase stages with concomitant decrease in percent mitotic index in the methanol extract (~5.64), fraction, and patuletin (~4) as compared to the control (~7.61). However, in only methanol extract-treated root tips, an increase in metaphase stage was noted. In addition, the methanol extract predominantly induced c-type, misaligned, and multipolar chromosomal abnormalities while the fraction and patuletin displayed fragments and sticky chromosomes. The fraction and patuletin also produced micronuclei (~2%). In conclusion, T. patula flower methanol extract and ethyl acetate fraction are cytotoxicand genotoxic, which most likely could be due to the patuletin. Further preclinical and clinical studies are required to justify its clinical use.

Time-dependent impairments in learning and memory in Streptozotocin-induced hyperglycemic rats.

The sedentary lifestyle is responsible for the high prevalence of diabetes which also impairs cognition including learning and memory. Various studies have highlighted the learning and memory impairments in rodent models but data regarding the timeline of their development and their correlation to biochemical parameters are scarce. So, the present study was designed to investigate the type of memory which is more susceptible to hyperglycemia and its correlation with biochemical parameters such as inflammatory cytokines, cAMP response element binding (CREB) and protein kinase B (Akt) activation. Hyperglycemia was induced using streptozotocin (STZ, 45 mg/kg i.p.) and confirmed by measuring fasting blood glucose levels after 1 week of STZ injection. Learning and memory deficits were evaluated using the Novel Object Recognition Test (NORT) and Morris water maze (MWM), and correlated with biochemical parameters (TNF-α, IL-1ß, and dopamine) at 3, 6 and 9 weeks. STZ-injected rats after 3 weeks of injection demonstrated moderate hyperglycemia (blood glucose = 7.99 ± 0.62 mM) with intact learning and reference memory; however, their working memory was impaired in MWM. Severe hyperglycemia (blood glucose = 11.51 ± 0.69 mM) accompanied by impaired short, long, and working memory was evident after 6 weeks whereas learning was intact. After 9 weeks of STZ injection, hyperglycemia was more pronounced (13.69 ± 1.43 mM) and accompanied by a learning deficit in addition to short, long, and working memory impairments. The extent of hyperglycemia either in terms of duration or severity resulted in enhanced inflammation, down-regulation of the level of dopamine, protein expression of AKT and CREB, which possibly affected learning and memory negatively.

Comparative In Vitro Assessment of the Methanol Extracts of the Leaf, Stem, and Root Barks of Cnidoscolus aconitifolius on Lung and Breast Cancer Cell Lines.

OBJECTIVES: Cnidoscolus aconitifolius Mill. I.M.Johnst. is a medicinal plant widely used in ethnomedicine for the treatment of cancer and other diseases. MATERIALS AND METHODS: The effects of methanol extracts of the leaf, stem, and root barks were evaluated on breast (MCF-7) and lung (NCI-H460) cancer cells at 1-250 µg/mL using the SRB assay and the extracts were screened for phytochemicals using the standard method. RESULTS: The stem and root extracts showed no activity at the maximum concentration, while the leaf extract at 100 µg/mL showed remarkable cell growth inhibition against breast (-14.50±0.58) and lung cancer (+53.29±4.57) in vitro. The extracts showed the presence of saponins, terpenes, cardiac glycosides, and phenolic compounds. Partitioning of the active leaf extract further enhanced its activity as the chloroform fraction exhibited GI50, LC50, and total growth inhibition (TGI) of 22.5, 68.75, and 43.75 µg/mL against breast cancer, respectively, and GI50 and TGI of 35.4 and 55.8 µg/ mL against lung cancer cells, respectively. However, the aqueous fraction showed no cytotoxicity against either cell line. CONCLUSION: These results justified the ethnomedicinal uses of the plant against tumor-related ailments. Isolation of the constituents responsible for the observed activity needs to be carried out to further support this claim.

Cytotoxic, embryotoxic, insecticidal and anti-microbial activities of standardized Areca catechu nut.

The study was aimed at evaluating various biological actions of widely consumed Areca catechu nut. The nut's ethanolic extract exhibited cytotoxicity (lung cancer cell line), embryotoxicity (chick embryo), phytotoxicity (Lemna minor), insecticidal (Rhyzopertha dominica), anti-bacterial (Pseudomonas aeruginosa), anti-fungal (Microsporum canis) and mitogenic (human blood lymphocytes) actions. The standardization results revealed presence of 1.7 µ g arecoline per mg of extract. In conclusion, the Areca nut is endowed with both harmful and beneficial biological actions. Keeping in view its wide consumption and ease of availability, the aforesaid information should be channelized for health and agricultural benefits.

Depressive symptoms, monoamines levels, MAO-B activity and effect of treatment in a subset of depressed individuals from government sector hospital at Karachi.

Depression is one of the leading causes of disability in developing countries including Pakistan. This study was designed to assess the frequency and severity of depressive symptoms, monoamines and their metabolite levels, MAO-B activities before and after treatment with antidepressants in a sub-set of Karachi population in Pakistan. Drug naive depressed subjects were evaluated before and after treatment with selective serotonin reuptake inhibitors. Symptoms of depressed mood and anxiety psychic (90%) were more frequent whereas, suicidal thoughts (~50%) and feelings of guilt (~30%) were less common. Hamilton Depression Rating Scale scores were 21.4 ± 0.8 in both genders with a significantly higher score (1.3x) in females. Homovanillic acid, 5- hydroxyindoleacetic acid and MAO-B activity were significantly higher 43%, 66% and 25% respectively, in depressed than normal subjects. A significant decline after 2 weeks treatment in HDRS scores with fluoxetine (19%) and paroxetine (40%) and in MAO-B activity (20%) was observed. In conclusion, in our population early decline in HDRS scores supports that they are SSRIs responders, whereas a concomitant reduction in MAO-B activities indicates that it can be considered as one of the parameters for early detection of response. Additionally, the low frequency of suicidal thoughts could be associated with higher levels of monoamine metabolites.

Antibiofilm potential of 16-oxo-cleroda-3, 13(14) E-diene-15 oic acid and its five new γ-amino γ-lactone derivatives against methicillin resistant Staphylococcus aureus and Streptococcus mutans.

The clerodane diterpenoids 16-oxo-cleroda-3, 13(14) E-diene-15 oic acid (1) and kolavenic acid (2) isolated from Polyalthia longifolia var. pendula (Linn.) were previously reported for their antimicrobial activity. Thus present study was designed to investigate the biofilm inhibiting potential of these diterpenoids (1-2) and five new lactone derivatives (3-7) of 1 against methicillin resistant Staphylococcus aureus (MRSA), Streptococcus mutans, Klebsiella pneumoniae and Proteus mirabilis. Compounds 1 and 3 at 10-20 µg/mL were found to be bacteriostatic and significantly reduced the biofilm formation and metabolically active cells of MRSA and S. mutans up to 90%. Parental diterpenoid (1) at 10 and 16 µg/mL significantly eradicated the preformed biofilm of both pathogens. Both the compounds also delayed acid production at minimum inhibitory (MIC) and sub-inhibitory concentrations (sub MIC). Florescence and scanning electron microscopy further confirms the biofilm inhibiting potential of compounds 1 and 3 and displayed disrupted biofilms at MIC and sub MIC levels. The gene expression of MRSA and S. mutans responsible for biofilm formation was also altered. Moreover, the observed anti-virulence properties and delayed bacterial growth after 10 min of exposure to the test compounds 1 and 3 make them a promising class of antibiofilm agents.

Cytotoxicity and chromosomal aberrations induced by methanolic extract of Cuscuta reflexa and its pure compounds on meristematic cells of Allium species.

Cuscuta reflexa (Convolvulaceae), is commonly known as amarbel or akashbel. In Bangladesh and Nepal some of the tribes use C. reflexa against edema, body ache, cancer, skin infections and liver disorders. Despite its traditional uses there is no information regarding genotoxic effects of either the plant extract or its pure compounds. Methanolic extract of C. reflexa (MECR) and pure compounds derived from it namely, odoroside H, neritaloside, and strospeside, were evaluated in Allium cepa L. and A. sativum L. for their effects on root growth, root apical meristem mitotic index (MI) , and chromosomal aberrations (CAs). In this study, we adopted a new method of calculating percent change in root length. MECR caused a concentration- and time- dependent inhibition in root length at 100 - 10000µg/ml in A. cepa root. It was accompanied by a subsequent decline in MI which is an indicative of its cytotoxic effect. On the contrary, at low concentrations a significant rise in root length was noticeable. In A. sativum, MECR also reduced the root length having IC50 values ~8 x and 4.3 x lower than A. cepa. A variety of CAs were evident in both Allium systems after treatment with MECR, odoroside H and neritaloside. Thus in MECR, cardenolides glycosides, i.e. odoroside H and neritaloside could be accountable for its genotoxicity.

In vitro growth inhibition and cytotoxicity of Euphorbia caducifolia against four human cancer cell lines and its phytochemical characterisation.

Several Euphorbia species have been used in folklore as cancer remedies, however, scientific studies on the cytotoxicity (in vitro studies) of Euphorbia caducifolia are lacking. In present study, anticancer potential of E. caducifolia aerial parts ethanol extract and its fractions were evaluated against human lung (NCI-H460), breast (MCF-7), prostate (PC-3) and cervical (HeLa) cancer cell lines, using sulphorhodamine-B in vitro cytotoxicity (in vitro studies) assay. The ethanol extract demonstrated growth inhibitory effect against all aforementioned cancer cell lines with IC50, 19-135 µg/mL and LC50, ~220 µg/mL, and its petroleum ether fraction obtained on bioactivity guided fraction showed highest activity with IC50, 28-70 µg/mL and LC50, 71 µg/mL against NCI-H460 and MCF-7 cell lines. Its phytochemicals were analysed by gas chromatography-mass spectrometry (GC-MS). The present study provides scientific justification for its traditional use against cancer.

Opuntia dillenii cladode: Opuntiol and opuntioside attenuated cytokines and eicosanoids mediated inflammation.

ETHANOPHARMACOLOGICAL RELEVANCE: Opuntia dillenii Haw (Nagphana) traditionally used against inflammation. The present study addressed the anti-inflammatory activity of O. dillenii derived methanol extract, fractions and pure compounds and their underlying mechanism of action. MATERIALS AND METHODS: O. dillenii cladode methanol extract was subjected to vacuum liquid chromatography (VLC) furnishing two main fractions viz (T-1 and -2) leading to isolation of opuntiol (aglycone) and opuntioside (O-glucoside), respectively. Anti-inflammatory activity of extract, fractions, pure compounds and reference drugs were evaluated using: (1) arachidonic acid (AA) and 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced ear edema accompanied by histological studies of mice ear sections and phospholipase A2 (PLA2)-induced mice paw edema. (2) Carrageenan and glycogen-induced peritonitis in rodents. In parallel levels of leukotriene B4 (LTB4) and reactive oxygen species (ROS) were also determined via HPLC and fluoroemetrically using 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA) dye, respectively. Additionally, levels of prostaglandin E2 (PGE2), tumor necrosis factor (TNF-α), interleukins IL-1ß and -6 were measured by ELISA assay. RESULTS: O. dillenii methanol extract, fractions and pure compounds reduced AA and TPA-induced ear punch weight in a dose dependent fashion. The corresponding IC50 values obtained also suppressed inflammatory features observed histologically. Furthermore, paw edema and peritonitis were also attenuated. Similar to indomethacin and diclofenac sodium, opuntioside reduced PGE2 levels of inflamed ear which was comparatively 1.3× better than opuntiol. However, opuntiol was more potent in reducing LTB4 levels in rat neutrophils with an IC50 value of 19±3.3µΜ, while opuntioside was ineffective. Opuntiol also effectively suppressed ROS (37%) and cytokine levels (TNF-α, IL-1ß and -6) by ~50% and comparable to dexamethasone. CONCLUSIONS: O. dillenii cladodes possess anti-inflammatory properties via inhibition of arachidonic acid metabolites and cytokines. Opuntiol (aglycone) emerged as a dual inhibitor of cyclooxygenase (COX) and lipooxygenase (LOX) pathways. It also suppressed ROS and cytokine levels. However, opuntioside manifested its selectivity towards COX (PGE2) pathway without affecting LTB4 levels. The present report describing the anti-inflammatory activity of opuntiol and opuntioside for the first time thereby, supporting and justifying the traditional use of O. dillenii against inflammation and may serve as lead compound in designing of new anti-inflammatory agents.

Investigation of cyclooxygenase and signaling pathways involved in human platelet aggregation mediated by synergistic interaction of various agonists.

In the present study, the mechanism(s) of synergistic interaction of various platelet mediators such as arachidonic acid (AA) when combined with 5-hydroxytryptamine (5-HT) or adenosine diphosphate (ADP) on human platelet aggregation were examined. The results demonstrated that 5-HT had no or negligible effect on aggregation but it did potentiate the aggregation response of AA. Similarly, the combination of subeffective concentrations of ADP and AA exhibited noticeable rise in platelet aggregation. Moreover, the observed synergistic effect of AA with 5-HT on platelets was inhibited by different cyclooxygenase (COX) inhibitors, namely ibuprofen and celecoxib, with half maximal inhibitory effect (IC50) values of 18.0 ± 1.8 and 15.6 ± 3.4 µmol/L, respectively. Interestingly, the synergistic effect observed for AA with 5-HT was, also, blocked by the 5-HT receptor blockers cyproheptadine (IC50=22.0 ± 7 µmol/L), ketanserin (IC50=152 ± 23 µmol/L), phospholipase C (PLC) inhibitor (U73122; IC50=6.1 ± 0.8 µmol/L), and mitogen activated protein kinase (MAPK) inhibitor (PD98059; IC50=3.8 ± 0.5 µmol/L). Likewise, the synergism of AA and ADP was, also, attenuated by COX inhibitors (ibuprofen; IC50=20 ± 4 µmol/L and celecoxib; IC50=24 ± 7 µmol/L), PLC inhibitor (U73122; IC50=3.7 ± 0.3 µmol/L), and MAPK inhibitor (PD98059; IC50=2.8 ± 1.1 µmol/L). Our observed data demonstrate that the combination of subthreshold concentrations of agonists amplifies platelet aggregation and that these synergistic effects largely depend on activation of COX/thromboxane A2, receptor-operated Ca(2+) channels, Gq/PLC, and MAPK signaling pathways. Moreover, our data revealed that inhibition of COX pathways by using both selective and/or non-selective COX inhibitors blocks not only AA metabolism and thromboxane A2 formation, but also its binding to Gq receptors and activation of receptor-operated Ca(2+) channels in platelets. Overall, our results show that PLC and MAPK inhibitors proved to inhibit the synergistic activation of platelets by several/multiple agonists.

Cytotoxic and antioxidant properties of phenolic compounds from Tagetes patula flower.

CONTEXT: Tagetes patula Linn. (Asteraceae) (French Marigold) flowers are used by local practitioners for cancer treatment; however, it lacks scientific justification. OBJECTIVE: Identification of bioactive compounds in T. patula flower for cytotoxic and growth inhibition in human cancer cell lines along with its antioxidant properties using chemical and cell based systems. MATERIALS AND METHODS: The T. patula flower methanol extract, its seven fractions, and three phenolic compounds including methyl protocatechuate (1), patuletin (2), and patulitrin (3) were evaluated using sulforhodamine-B assay against HeLa, HT-144, NCI-H460, MCF-7, PC-3, and SF-268 human cancer cell lines. In parallel, antioxidant activity was evaluated using chemical (DPPH(·), deoxyribose, and lipid peroxidation assays) and cell-based chemiluminescence systems (human neutrophils and mice macrophages). RESULTS: The methanol extract and ethyl acetate insoluble fraction exhibited cytotoxic and growth inhibitory effects against HeLa in which 2 exhibited highest cell growth inhibition (GI50: 0.6 ± 0.1 µg/ml) and cytotoxicity (LC50: 2.5 ± 0.1 µg/ml). It also scavenged LOO(·) (IC50: 6.5 ± 0.7 µg/ml) and [Formula: see text] (IC50: 27.5 ± 1.3 µg/ml) in chemical systems and human neutrophils, respectively. However, 1 preferably scavenged H2O2-Cl(-) (IC50: 0.5 ± 0.01 µg/ml) in mice macrophages. DISCUSSION AND CONCLUSION: Compound 2 from T. patula flower exhibited both growth inhibitory and cytotoxic properties while 1 and 3 were only growth inhibitory against HeLa. 1-3 also displayed antioxidant properties implying its probable role in growth inhibition/cytotoxic action. The present study provides scientific evidence for the use of T. patula flower in cancer treatment by traditional healer.

Anticancer and antioxidant activities of methanol extracts and fractions of some Cameroonian medicinal plants.

OBJECTIVE: To obtain a scientific basis of the use of plant-derived preparations by many rural people in Cameroon, for their primary health care needs in the treatment of diseases such as cancer. METHODS: The antiproliferative effect of 11 plants methanol crude extracts on four cancer cells using sulforhodamine-B assay and their antioxidant activities using 1-diphenyl-2-picrylhydrazyl radical and nitric oxide radical scavenging ability were investigated. The Ekebergia senegalensis (E. senegalensis) and Protea elliotii (P. elliotii) extracts were selected based on their antioxidant and anticancer activities, and partition in hexane, dichloromethane, ethyl acetate, butanol and methanol was done. Each fraction was submitted to antioxidant and anticancer activities, and the effect of the dichloromethane fraction (the most antiproliferative fraction) on NCI-H460 cell cycle was determined by flow cytometry. RESULTS: The most antiproliferative substances were found for the extracts from E. senegalensis, P. elliotii, Terminalia macroptera and Vitellaria paradoxa. Whereas the most antioxidant substances were found for the extracts from Cissus populnea, E. senegalensis, P. elliotii, Terminalia macroptera, Vitellaria paradoxa, and Gardenia aqualla. Dichloromethane fraction of P. elliotii was found to be highly antiproliferative to NCI-H460 cancer cells and showed S phase arrest cell cycle progression. Ethyl acetate n-butanol and methanol fractions showed quite strong antioxidant activity for both E. senegalensis and P. elliotii, as compared to that of gallic acid. CONCLUSIONS: Overall, the antiproliferative and antioxidant activities of some of the extracts lend some support to their use in the traditional medicine of Adamawa Region, Cameroon to treat cancer.

Neuroprotective effects of forsythiaside on learning and memory deficits in senescence-accelerated mouse prone (SAMP8) mice.

Forsythiaside (3,4-dihydroxy-ß-phenethyl-O-α-L-rhamnopyranosyl-(1→6)-4-O-caffeo yl-ß-d-glucopyranoside, C29H36O15), which is isolated from air-dried fruits of Forsythia suspensa, has been shown to possess anti-oxidant, anti-bacterial and anti-inflammatory activities. The aim of this study is to investigate the neuroprotective effects of forsythiaside on learning and memory deficits in the senescence-accelerated mouse prone 8 (SAMP8, a model of age-dependent neurodegenerative disorders such as Alzheimer's disease). Forsythiaside (60, 120 and 240mg/kg) was orally administered to aged (8months old) SAMP8 mice for 45days followed by evaluating cognitive impairment (Morris water maze and step-through passive avoidance), inflammation (interleukin-1ß (IL-1ß) and interleukin-6 (IL-6) levels), oxidative stress (glutathione peroxidase (GSH-Px) and total superoxide dismutase (T-SOD) activities; malondialdehyde (MDA) and nitric oxide (NO) contents) and neurotransmitter such as norepinephrine (NE), dopamine (DA), 5-hydroxytryptamine (5-HT), glutamate (GLU) gamma-aminobutyric acid (GABA) and acetyl choline (ACh). In Morris water maze, forsythiaside had significantly reduced the latency time, the crossing numbers and time spent in target quadrant compared to aged SAMP8 mice. In passive avoidance test, a significant decline in number of errors while increase in latency was observed when compared with aged SAMP8 mice. Furthermore, a significant decrease in IL-1ß, NO, MDA and NE levels, and an increase in T-SOD and GSH-Px activities and GLU and Ach levels were evident in the brain homogenates of forsythiaside-treated mice compared to aged SAMP8 mice. These findings demonstrated that forsythiaside may be a useful treatment against amnesia.

Flavonoid and cardenolide glycosides and a pentacyclic triterpene from the leaves of Nerium oleander and evaluation of cytotoxicity.

A pentacyclic triterpene, oleanderocioic acid, two flavonoidal glycosides, quercetin-5-O-[α-L-rhamnopyranosyl-(1→6)]-ß-D-glucopyranoside and kaempferol-5-O-[α-L-rhamnopyranosyl-(1→6)-ß-D-glucopyranoside, and a cardenolide, oleandigoside, together with 11 known compounds, were isolated from the leaves of Nerium oleander. Their structures were elucidated on the basis of spectroscopic analysis. The growth inhibitory and cytotoxic activities of eight compounds were evaluated against the MCF-7 human breast cancer cell line using a sulforhodamine B assay. Three compounds, oleandrin, odoroside A and B were further assayed using a panel of 57 human cancer cell lines.