Evaluation of Solanum linnaeanum and S. sisymbriifolium extracts for the management of Meloidogynechitwoodi.

Meloidogyne chitwoodi causes significant yield losses in many crops and the chemical control measures currently used are less effective for this nematode. The activity of aqueous extracts (0.8 mg/mL) of one-month-old (R1M) and two-months-old roots and immature fruits (F) of Solanum linnaeanum (Sl) and S. sisymbriifolium cv. Sis 6001 (Ss) were tested on hatching, mortality, infectivity and reproduction of M. chitwoodi. The extracts selected reduced the hatching of second-stage juveniles (J2) (cumulative hatching of 40% for Sl R1M and 24% for Ss F) but did not affect J2 mortality. However, infectivity of J2 exposed to the selected extracts, during 4 and 7 days, was lower (3% and 0% for Sl R1M and 0% and 0% for Ss F) compared to the control (23% and 3%). Reproduction was affected only after 7 days of exposure (reproduction factor (RF) was 7 for Sl R1M and 3 for Ss F) compared to the control (RF = 11). The results suggest that the selected Solanum extracts are effective and can be a useful tool in sustainable M. chitwoodi management. This is the first report on the efficacy of S. linnaeanum and S. sisymbriifolium extracts against root-knot nematodes.

Acanthus mollis Formulations for Transdermal Delivery: From Hydrogels to Emulsions.

Topical formulations of Acanthus mollis L. leaf and the optimization of the release of their active compounds and their topical bioavailability were investigated for the first time. In vitro, the release of active compounds from three formulations-an oil-in-water cream and two hydrogels (Carbopol 940 and Pluronic F-127)-was determined using Franz diffusion cells. Detection and quantification of the compounds was performed via high-performance liquid chromatography with a photodiode array (HPLC-PDA). DIBOA, a bioactive compound of this medicinal plant, exhibited release kinetics of the Weibull model for the Carbopol and Pluronic F-127 formulation, identifying it as a potential active agent to optimize the topical distribution of the formulations. The implications extend to applications in inflammation treatment and tyrosinase inhibition, suggesting that it can make a significant contribution to addressing skin conditions, including melanoma and various inflammatory diseases.

Synergistic Effect of DIBOA and Verbascoside from Acanthus mollis Leaf on Tyrosinase Inhibition.

Overexpression of melanin contributes to darkening of plant and fruit tissues and skin hyperpigmentation, leading to melasma or age spots. Although melanin biosynthesis is complex and involves several steps, a single enzyme known as tyrosinase is key to regulating this process. The melanogenesis pathway is initiated by oxidation of the starting material l-tyrosine (or l-DOPA) to dopaquinone by tyrosinase; the resulting quinone then serves as a substrate for subsequent steps that eventually lead to production of melanin. Medicinal plants are considered a good source of tyrosinase inhibitors. This study investigated the tyrosinase inhibitory activity of A. mollis leaf extracts and their phytochemicals. Significant activity was verified in the ethanol extract -EEt (IC50 = 1.21 µg/mL). Additionally, a kinetic study showed that this tyrosinase inhibition occurs by DIBOA (2,4-dihydroxy-1,4-benzoxazin-3-one) and verbascoside contribution through a non-competitive reaction mechanism. A synergistic effect on tyrosinase inhibition was observed in the binary combination of the compounds. In conclusion, both EEt and a mixture of two of its phytochemicals can be effective tyrosinase inhibitors and can be used as a bleaching agent for cosmetic formulations in the future.

Effect of Phenolic Compounds from Cymbopogon citratus (DC) Stapf. Leaves on Micellar Solubility of Cholesterol.

Dyslipidemias are one of the risk factors for cardiovascular diseases, the leading cause of death and hospitalization worldwide. One way to control cholesterol levels is to control the exogenous cholesterol intake in the body. Natural polyphenolic compounds, namely theaflavins from plant extracts such as black tea, showed the ability to inhibit the formation of the micellar structure, essential for the absorption of cholesterol in the intestine. There are several methodologies to determine this effect, many of which are expensive and time-consuming. Due to these facts, the main purposes of this work were to optimize an inexpensive colorimetric method to study, in vitro, the micellar solubility of cholesterol and applied it to plant extracts. In this work, Cymbopogon citratus leaf extracts, its phenolic fractions, and flavonoids were evaluated. The non-delipidified infusion (CcI) obtained a maximum percentage of micelle destruction of 59.22% for a concentration of 50 µg/mL and the delipidified infusion (CcdI) obtained a maximum percentage of micelle destruction of 58.01% for a concentration of 200 µg/mL. In the case of the fraction of phenolic acids (CcPAs), 23.85% of maximum micellar destruction was recorded for the concentration of 100 µg/mL, while for the fraction of flavonoids (CcF), the micellar destruction was 92.74% at 1 µg/mL, and for the tannin fraction (CcT) of 99.45% at 25 µg/mL. Luteolin presented a percentage of micelle destruction of 94.83% in the concentration of 1 ng/mL, followed by luteolin-7-O-glucoside with 93.71% and luteo-lin-6-C-glucoside with 91.26% at the concentrations of 25 ng/mL and 50 ng/mL, respectively. These results suggest the capability of polyphenols from Cymbopogon citratus to prevent the cholesterol absorption in the gut by micellar destruction, and its contribution for cholesterol-lowering activity.

Forest Biomass as a Promising Source of Bioactive Essential Oil and Phenolic Compounds for Alzheimer's Disease Therapy.

Alzheimer's disease (AD) is the most common neurodegenerative disorder affecting elderly people worldwide. Currently, there are no effective treatments for AD able to prevent disease progression, highlighting the urgency of finding new therapeutic strategies to stop or delay this pathology. Several plants exhibit potential as source of safe and multi-target new therapeutic molecules for AD treatment. Meanwhile, Eucalyptus globulus extracts revealed important pharmacological activities, namely antioxidant and anti-inflammatory properties, which can contribute to the reported neuroprotective effects. This review summarizes the chemical composition of essential oil (EO) and phenolic extracts obtained from Eucalyptus globulus leaves, disclosing major compounds and their effects on AD-relevant pathological features, including deposition of amyloid-ß (Aß) in senile plaques and hyperphosphorylated tau in neurofibrillary tangles (NFTs), abnormalities in GABAergic, cholinergic and glutamatergic neurotransmission, inflammation, and oxidative stress. In general, 1,8-cineole is the major compound identified in EO, and ellagic acid, quercetin, and rutin were described as main compounds in phenolic extracts from Eucalyptus globulus leaves. EO and phenolic extracts, and especially their major compounds, were found to prevent several pathological cellular processes and to improve cognitive function in AD animal models. Therefore, Eucalyptus globulus leaves are a relevant source of biological active and safe molecules that could be used as raw material for nutraceuticals and plant-based medicinal products useful for AD prevention and treatment.

A review of the ethnomedicinal uses, chemistry, and pharmacological properties of the genus Acanthus (Acanthaceae).

ETHNOPHARMACOLOGICAL RELEVANCE: The Acanthus genus belongs to the Acanthaceae family, and its species are distributed in all continents, mainly in tropical and subtropical regions. Several traditional applications are referred to, but few scientific studies validate them. Despite this, studies in animal models corroborate some of its uses in folk medicine, such as anticancer, antidiabetic, anti-inflammatory, and antinociceptive, which encourages the research on plants of this genus. AIM OF THE REVIEW: To our knowledge, this document is the first comprehensive review study that provides information on the geographic distribution, botanical characteristics, ethnomedicinal uses, phytochemicals, and pharmacological activities of some Acanthus species to understand the correlation between traditional uses, phytochemical, and pharmacological activities, providing perspectives for future studies. RESULTS: In traditional medicine, Acanthus species are mainly used for diseases of respiratory, nervous and reproductive system, gastrointestinal and urinary tract, and skin illness. The most used species are A. montanus, A. ilicifolius, and A. ebracteatus. Chemical compounds (125) from different chemical classes were isolated and identified in seven species, mainly from A. ilicifolius, about 80, followed by A. ebracteatus and A. montanus, appearing with a slightly lower number with fewer phytochemical profile studies. Isolated phytoconstituents have been mainly alkaloids, phenylpropanoid glycosides, and phenylethanoids. In addition, aliphatic glycosides, flavonoids, lignan glycosides, megastigmane derivatives, triterpenoids, steroids, fatty acids, alcohols, hydroxybenzoic acids, simple phenols were also cited. Scientific studies from Acanthus species extracts and their phytoconstituents support their ethnomedical uses. Antimicrobial activity that is the most studied, followed by the antioxidant, anti-inflammatory, and anticancer properties, underlie many Acanthus species activities. A. dioscoridis, A. ebracteatus, A. hirsutus, A. ilicifolius, A. mollis, A. montanus, and A. polystachyus have studies on these activities, A. ilicifolius being the one with the most publications. Most studies were essentially performed in vitro. However, the anticancer, antidiabetic, anti-inflammatory and antinociceptive properties have been studied in vivo. CONCLUSION: Acanthus species have remarkable phytoconstituents with different biological activities, such as antioxidant, antimicrobial, anti-inflammatory, antinociceptive, hepatoprotective, and leishmanicidal, supporting traditional uses of some species. However, many others remain unexplored. Future studies should focus on these species, especially pharmacological properties, toxicity, and action mechanisms. This review provides a comprehensive report on Acanthus genus plants, evidencing their therapeutic potential and prospects for discovering new safe and effective drugs from Acanthus species.

Chemical Composition and Effect against Skin Alterations of Bioactive Extracts Obtained by the Hydrodistillation of Eucalyptus globulus Leaves.

Eucalyptus globulus is planted extensively for pulp, paper and wood production. Although bioactive compounds obtained from its biomass are used as cosmetics ingredients, the skin effects were not yet fully explored. In order to fill this gap, this work aimed to study the protective effect against skin damage provided by the essential oil (EO) obtained from the hydrodistillation of Eucalyptus globulus leaves, and by an extract obtained from the hydrodistillation residual water (HRW). The major compound identified in the EO was 1,8-Cineole, and the phenolic acids in the HRW included gallic acid as the main phenolic constituent. Moreover, non-toxic EO and HRW concentrations were shown to have anti-aging skin effects in vitro, decreasing age-related senescence markers, namely ß-galactosidase and matrix metalloproteinases activation, as well as collagen type 1 upregulation. In addition, EO and HRW were found to exhibit depigmenting effects by inhibiting tyrosinase and melanin production, along with potent anti-inflammatory properties. Furthermore, the absence of skin irritation and sensitization in cells exposed to EO and HRW revealed the safety of both extracts for topical use. Taken together, these results highlight the beneficial effects of extracts obtained from Eucalyptus globulus biomass for skin aesthetic and health purposes, which should be explored deeply for the prediction of future pharmaceutical and dermocosmetics industrial applications.

Formulation Effects in the Antioxidant Activity of Extract from the Leaves of Cymbopogon citratus (DC) Stapf.

Cymbopogon citratus DC (Stapf.) is a perennial grass and it is distributed around the world. It is used as a condiment for food and beverage flavouring in the form of infusions and decoctions of its dried leaves. Our previous studies have shown antioxidant, anti-inflammatory and gastroprotective activities for the infusion and its phenolic fractions. The aim of the present work was to develop oral dosage forms from a Cymbopogon citratus extract to be used as a functional food with antioxidant properties. Initially, an essential oil-free infusion was prepared, lyophilized and characterized by HPLC-PDA. Total phenols were quantified with the Folin-Ciocalteu method and the antioxidant activity was assessed by DPPH assay. Gelatine capsules containing the extract with different excipients, selected after DSC and IR trials, were prepared. A formulation exhibiting better antioxidant behaviour in a gastric environment was attained. These results suggest that the proposed formulation for this extract could be a valuable antioxidant product and, consequently, make an important contribution to "preventing" and minimizing diseases related to oxidative stress conditions.

Solanum linnaeanum and Solanum sisymbriifolium as a sustainable strategy for the management of Meloidogyne chitwoodi.

Root-knot nematodes (RKN), Meloidogyne spp., are important crop pests that cause severe losses in crop production worldwide, reducing both productivity and crop quality. Meloidogyne chitwoodi Golden, O'Bannon, Santo & Finley, 1980 is considered a quarantine organism by the European and Mediterranean Plant Protection Organization (EPPO) causing damage in tomato and potato crops. The development of nonchemical and sustainable management strategies to reduce nematode damage is crucial. The resistance of Solanum linnaeanum Hepper & P.-M.L. Jaeger and S. sisymbriifolium Lamarck cv. Sis 6001 to M. chitwoodi was evaluated based on gall index (GI), the Bridge & Page (1980) rating chart and reproduction factor (RF). Both plant species were resistant to M. chitwoodi. Solanum linnaeanum had an average of 519 small root swellings/plant, with 45% adult nematodes inside the roots, all males. Solanum sisymbriifolium had GI ≤ 2 and RF ≤ 1 with a high percentage (69%) of nematodes inside the roots that did not develop beyond the sexually undifferentiated second-stage. The use of S. linnaeanum as a new source of resistance is a good alternative for the control of RKN in the quest to develop nonchemical and sustainable management strategies to protect crops.

Fragaria vesca L. Extract: A Promising Cosmetic Ingredient with Antioxidant Properties.

Fragaria vesca L. (F. vesca), popularly known as wild strawberry, is a plant from the Rosaceae family, found in temperate and subtropical areas of the northern hemisphere. F. vesca leaves have been shown to have antiseptic, emollient, and dermatological protection properties, due to the presence of bioactive compounds, such as flavonoids, phenolic acids, ellagitannins, and proanthocyanidins. In this study, a F. vesca extract was obtained by an optimized extraction process, and was characterized by HPLC, ROS scavenging activity, cytotoxicity assays in HaCaT cells, and tyrosinase inhibitory activity determination. The most active extract was then incorporated in a hydrogel with hydroxyethylcellulose at 2% (w/w), which was characterized at the physicochemical, stability, cytotoxicity, and ROS scavenging activity levels to evaluate its quality, safety, and efficacy. In vivo studies, human repeat insult patch testing, and an assay to determine their antioxidant efficacy, were also performed. The results showed that the Fragaria vesca extracts had antioxidant activity and that the F. vesca extract-based hydrogel exhibited cutaneous compatibility, acceptability and antioxidant efficacy, being stable, and suitable for topical application.

Acanthus mollis L. leaves as source of anti-inflammatory and antioxidant phytoconstituents.

This work expands the phytochemical composition knowledge of Acanthus mollis and evaluates antioxidant and anti-inflammatory activities which could be related with its traditional uses. Extracts from leaves, obtained by sequential extraction, were screened using TLC and HPLC-PDA. The ethanol extract was the most active on DPPH assay (IC50 = 20.50 µg/mL) and inhibited nitric oxide (NO) production in RAW 264.7 macrophages (IC50 = 48.31 µg/mL). Significant amounts of cyclic hydroxamic and phenolic acids derivatives were detected. A lower antioxidant effect was verified for a fraction enriched with DIBOA derivatives (IC50 = 163.02 µg/mL), suggesting a higher contribution of phenolic compounds for this activity in ethanol extract. However, this fraction exhibited a higher inhibition of NO production (IC50 = 32.32 µg/mL), with absence of cytotoxicity. These results support the ethnomedical uses of this plant for diseases based on inflammatory processes. To our knowledge, it is the first report to the anti-inflammatory activity for DIBOA derivatives.

Urtica spp.: Phenolic composition, safety, antioxidant and anti-inflammatory activities.

Urtica dioica and other less studied Urtica species (Urticaceae) are often used as a food ingredient. Fifteen hydroxycinnamic acid derivatives and sixteen flavonoids, flavone and flavonol-type glycosides were identified in hydroalcoholic extracts from aerial parts of Urtica dioica L., Urtica urens L. and Urtica membranacea using HPLC-PDA-ESI/MSn. Among them, the 4-caffeoyl-5-p-coumaroylquinic acid and three statin-like 3-hydroxy-3-methylglutaroyl flavone derivatives were identified for the first time in Urtica urens and U. membranacea respectively. Urtica membranacea showed the higher content of flavonoids, mainly luteolin and apigenin C-glycosides, which are almost absent in the other species studied. In vitro, Urtica dioica exhibited greater antioxidant activity but Urtica urens exhibited stronger anti-inflammatory potential. Interestingly, statin-like compounds detected in Urtica membranacea have been associated with hypocholesterolemic activity making this plant interesting for future investigations. None of the extracts were cytotoxic to macrophages and hepatocytes in bioactive concentrations (200 and 350µg/mL), suggesting their safety use in food applications.

Urolithins impair cell proliferation, arrest the cell cycle and induce apoptosis in UMUC3 bladder cancer cells.

Ellagitannins have been gaining attention as potential anticancer molecules. However, the low bioavailability of ellagitannins and their extensive metabolization in the gastrointestinal tract into ellagic acid and urolithins suggest that the health benefits of consuming ellagitannins rely on the direct effects of their metabolites. Recently, chemopreventive and chemotherapeutic activities were ascribed to urolithins. Nonetheless, there is still a need to screen and evaluate the selectivity of these molecules and to elucidate their cellular mechanisms of action. Therefore, this work focused on the antiproliferative effects of urolithins A, B and C and ellagic acid on different human tumor cell lines. The evaluation of cell viability and the determination of the half-maximal inhibitory concentrations indicated that the sensitivity to the studied urolithins varied markedly between the different cell lines, with the bladder cancer cells (UMUC3) being the most susceptible. In UMUC3 cells, urolithin A was the most active molecule, promoting cell cycle arrest at the G2/M checkpoint, increasing apoptotic cell death and inhibiting PI3K/Akt and MAPK signaling. Overall, the present study emphasizes the chemopreventive/chemotherapeutic potential of urolithins, highlighting the stronger effects of urolithin A and its potential to target transitional bladder cancer cells.

Polyphenols from Cymbopogon citratus leaves as topical anti-inflammatory agents.

ETHNOPHARMACOLOGICAL RELEVANCE: A variety of plant polyphenols have been reported to have anti-inflammatory, frequently associated with erythema, edema, hyperplasia, skin photoaging and photocarcinogenesis. Cymbopogon citratus (DC). Stapf (Poaceae) is a worldwide known medicinal plant, used in traditional medicine in inflammation-related conditions. AIM OF THE STUDY: In this work, the anti-inflammatory potential of C. citratus infusion (CcI) and its polyphenols as topical agents was evaluated in vivo. MATERIALS AND METHODS: The plant extract was prepared and its fractioning led two polyphenol-rich fractions: flavonoids fraction (CcF) and tannins fraction (CcT). An oil/water emulsion was developed with each active (CcI, CcF+CcT and diclofenac), pH and texture having been evaluated. Release tests were further performed using static Franz diffusion cells and all collected samples were monitored by HPLC-PDA. In vivo topical anti-inflammatory activity evaluation was performed by the carrageenan-induced rat paw edema model. RESULTS: The texture analysis revealed statistically significant differences for all tested parameters to CcF+CcT, supporting its topical application. Release experiments lead to the detection of the phenolic compounds from each sample in the receptor medium and the six major flavonoids were quantified, by HPLC-PDA: carlinoside, isoorientin, cynaroside, luteolin 7-O-neohesperidoside, kurilesin A and cassiaoccidentalin B. The CcF+CcT formulation prompted to the higher release rate for all these flavonoids. CcI4%, CcI1% and CcF+CcT exhibited an edema reduction of 43.18, 29.55 and 59.09%, respectively. CONCLUSIONS: Our findings highlight that CcI, containing luteolin 7-O-neohesperidoside, cassiaoccidentalin B, carlinoside, cynaroside and tannins have a potential anti-inflammatory topical activity, suggesting their promising application in the treatment of skin inflammatory pathologies.

Gastroprotective effect of Cymbopogon citratus infusion on acute ethanol-induced gastric lesions in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Treatment of gastric ulcers with medicinal plants is quite common in traditional medicine worldwide. Cymbopogon citratus (DC) Stapf. leaves infusion has been used in folk medicine of many tropical and subtropical regions to treat gastric disturbances. The aim of this study was to assess the potential gastroprotective activity of an essential oil-free infusion from C. citratus leaves in acute gastric lesions induced by ethanol in rat. MATERIALS AND METHODS: The study was performed on adult male Wistar rats (234.0±22.7g) fasted for 24h but with free access to water. The extract was given orally before (prevention) or after (treatment) intragastric administration of absolute ethanol. Effects of dose (28 or 56mg/kg of body weight) and time of contact of the extract with gastric mucosa (1 or 2h) were also assessed. Animals were sacrificed, being the stomachs removed and the lesions were assessed by macroscopic observation and histopathology. RESULTS: C. citratus extract, given orally before or after ethanol, significantly (P<0.01) reduced gastric mucosal injury compared with control group (vehicle+ethanol). The effect does not appear to be dose-dependent. Results also suggested that the extract is more effective when the time of contact with gastric mucosa increases. CONCLUSIONS: The results of this assay confirm the gastroprotective activity of C. citratus extract on experimental gastric lesions induced by ethanol, contributing for the pharmacological validation of its traditional use.

Flavan hetero-dimers in the Cymbopogon citratus infusion tannin fraction and their contribution to the antioxidant activity.

Cymbopogon citratus (lemongrass) leaf infusion, a commonly used ingredient in Asian, African and Latin American cuisines, is also used in traditional medicine for the treatment of several pathological conditions; however, little is known about their bioactive compounds. Recent studies revealed the crucial role of the phenolic compounds namely flavonoids and tannins on the infusion bioactivity. Flavonoids have already been characterized; however the tannin fraction of lemongrass infusion is still uncharted. The aim of the present work is to characterize this fraction, and to evaluate its contribution to the antioxidant potential of this plant. Chemical characterization was achieved by HPLC-DAD-ESI/tandem MS and the antioxidant activity was evaluated using DPPH, ABTS and FRAP assays. Hetero-dimeric flavan structures have been described for the first time in lemongrass consisting of apigeniflavan or luteoliflavan units linked to a flavanone, either naringenin or eriodictyol, which may occur as aglycone or glycosylated forms. The antioxidant capacity of the fraction containing these compounds was significantly higher than the infusion, indicating its potential as a source of natural antioxidants.

Cymbopogon citratus industrial waste as a potential source of bioactive compounds.

BACKGROUND: Cymbopogon citratus (Cc), commonly known as lemongrass, is a very important crop worldwide, being grown in tropical countries. It is widely used in the food, pharmaceutical, cosmetic and perfumery industries for its essential oil. Cc aqueous extracts are also used in traditional medicine. They contain high levels of polyphenols, which are known for their antioxidant and anti-inflammatory properties. Hydrodistillation of lemongrass essential oil produces an aqueous waste (CcHD) which is discarded. Therefore a comparative study between CcHD and Cc infusion (CcI) was performed to characterize its phytochemical profile and to research its antioxidant and anti-inflammatory potential. RESULTS: HPLC-PDA/ESI-MS(n) analysis showed that CcI and CcHD have similar phenolic profiles, with CcHD presenting a higher amount of polyphenols. Additionally, both CcI and CcHD showed antioxidant activity against DPPH (EC50 of 41.72 ± 0.05 and 42.29 ± 0.05 µg mL(-1) respectively) and strong anti-inflammatory properties, by reducing NO production and iNOS expression in macrophages and through their NO-scavenging activity, in a dose-dependent manner. Furthermore, no cytotoxicity was observed. CONCLUSION: The data of this study encourage considering the aqueous solution from Cc leaf hydrodistillation as a source of bioactive compounds, which may add great industrial value to this crop.

Bioactivity of Fragaria vesca leaves through inflammation, proteasome and autophagy modulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Fragaria vesca leaves have been used in folk medicine for the treatment of several diseases, namely gastrointestinal, cardiovascular and urinary disorders, which could be related with the potential anti-inflammatory properties of the extract. This work aims to disclose the bioactivity and the underlying action mechanism of an extract from Fragaria vesca leaves in order to support its traditional uses. MATERIALS AND METHODS: A hydroalcoholic extract was prepared from Fragaria vesca leaves and its anti-inflammatory potential was evaluated through inhibition of nitric oxide production and expression of several pro-inflammatory proteins in lipopolysaccharide-triggered macrophages. Nitric oxide scavenger activity was also assessed using a standard nitric oxide donor. Since numerous inflammatory proteins are tightly regulated by ubiquitination and proteasomal degradation, the putative effect of the extract on these cellular proteolytic pathways was also disclosed. The phytochemical characterization was performed by HPLC-PDA-ESI/MSn and compared with an infusion prepared according to the traditional method. RESULTS: For non-cytotoxic concentrations (80 and 160µg/mL) the extract inhibited nitrite production, probably due to a direct nitric oxide scavenging. Furthermore, inhibition of proteasome activity was verified, leading to accumulation of ubiquitinated proteins. The extract also increased the conversion of the microtubule-associated protein light chain LC3-I to LC3-II, a marker of autophagy. Polyphenols, namely ellagitannins, proanthocyanidins, and quercetin and kaempferol glucuronide derivatives were identified in Fragaria vesca leaves extract. Most of the identified phenolic compounds matched with those found in traditional preparation, the infusion. CONCLUSIONS: The extract has a direct nitric oxide scavenging activity giving support to the traditional use of this plant for the treatment of inflammatory disorders. Furthermore, the extract affects the proteolytic systems but its role in cancer treatment requires further studies.

Screening and identification of neuroprotective compounds relevant to Alzheimer׳s disease from medicinal plants of S. Tomé e Príncipe.

ETHNOPHARMACOLOGICAL RELEVANCE: Alzheimer׳s disease (AD) neuropathology is strongly associated with the activation of inflammatory pathways, and long-term use of anti-inflammatory drugs reduces the risk of developing the disease. In S. Tomé e Príncipe (STP), several medicinal plants are used both for their positive effects in the nervous system (treatment of mental disorders, analgesics) and their anti-inflammatory properties. The goal of this study was to determine whether a phenotypic, cell-based screening approach can be applied to selected plants from STP (Voacanga africana, Tarenna nitiduloides, Sacosperma paniculatum, Psychotria principensis, Psychotria subobliqua) in order to identify natural compounds with multiple biological activities of interest for AD therapeutics. MATERIALS AND METHODS: Plant hydroethanolic extracts were prepared and tested in a panel of phenotypic screening assays that reflect multiple neurotoxicity pathways relevant to AD-oxytosis in hippocampal nerve cells, in vitro ischemia, intracellular amyloid toxicity, inhibition of microglial inflammation and nerve cell differentiation. HPLC fractions from the extract that performed the best in all of the assays were tested in the oxytosis assay, our primary screen, and the most protective fraction was analyzed by mass spectrometry. The predominant compound was purified, its identity confirmed by ESI mass spectrometry and NMR, and then tested in all of the screening assays to determine its efficacy. RESULTS: An extract from the bark of Voacanga africana was more protective than any other plant extract in all of the assays (EC50s≤2.4 µg/mL). The HPLC fraction from the extract that was most protective against oxytosis contained the alkaloid voacamine (MW=704.90) as the predominant compound. Purified voacamine was very protective at low doses in all of the assays (EC50s≤3.4 µM). CONCLUSION: These findings validate the use of our phenotypic screening, cell-based assays to identify potential compounds to treat AD from plant extracts with ethnopharmacological relevance. Our study identifies the alkaloid voacamine as a major compound in Voacanga africana with potent neuroprotective activities in these assays.

Propolis and its constituent caffeic acid suppress LPS-stimulated pro-inflammatory response by blocking NF-κB and MAPK activation in macrophages.

ETHNOPHARMACOLOGICAL RELEVANCE: Propolis is a bee product with numerous biological and pharmacological properties, such as immunomodulatory and anti-inflammatory activities. It has been used in folk medicine as a healthy drink and in food to improve health and prevent inflammatory diseases. However, little is known about its mechanism of action. Thus, the goal of this study was to verify the antioxidant activity and to explore the anti-inflammatory properties of propolis by addressing its intracellular mechanism of action. Caffeic acid was investigated as a possible compound responsible for propolis action. MATERIALS AND METHODS: The antioxidant properties of propolis and caffeic acid were evaluated by using the 2,2-Diphenyl-1-picrylhydrazyl free radical (DPPH) scavenging method. To analyze the anti-inflammatory activity, Raw 264.7 macrophages were treated with different concentrations of propolis or caffeic acid, and nitric oxide (NO) production, a strong pro-inflammatory mediator, was evaluated by the Griess reaction. The concentrations of propolis and caffeic acid that inhibited NO production were evaluated on intracellular signaling pathways triggered during inflammation, namely p38 mitogen-activated protein kinase (MAPK), c-jun NH2-terminal kinase (JNK1/2), the transcription nuclear factor (NF)-κB and extracellular signal-regulated kinase (ERK1/2), through Western blot using specific antibodies. A possible effect of propolis on the cytotoxicity of hepatocytes was also evaluated, since this product can be used in human diets. RESULTS: Caffeic acid showed a higher antioxidant activity than propolis extract. Propolis and caffeic acid inhibited NO production in macrophages, at concentrations without cytotoxicity. Furthermore, both propolis and caffeic acid suppressed LPS-induced signaling pathways, namely p38 MAPK, JNK1/2 and NF-κB. ERK1/2 was not affected by propolis extract and caffeic acid. In addition, propolis and caffeic acid did not induce hepatotoxicity at concentrations with strong anti-inflammatory potential. CONCLUSIONS: Propolis exerted an antioxidant and anti-inflammatory action and caffeic acid may be involved in its inhibitory effects on NO production and intracellular signaling cascades, suggesting its use as a natural source of safe anti-inflammatory drugs.