Antinociceptive and anti-inflammatory properties of aqueous extract obtained from Serjania marginata Casar leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Serjania marginata Casar (Sapindaceae Family) Leaves are popularly used against abdominal pain. Antiulcer properties of S. marginata were scientifically described, however rare studies showed the antinociceptive effects of this plant. AIM OF STUDY: In this study, we investigated the antinociceptive and anti-inflammatory effects of aqueous extract obtained from Serjania marginata leaves (AESM) in nociception/inflammation models. MATERIAL AND METHODS: AESM was analyzed in FIA-ESI-IT-MS and Mass spectrometer LTQ XL. AESM oral administration (p.o.) (30, 100 and 300 mg/kg), dexamethasone subcutaneous injection (1 mg/kg, s.c.) and morphine (5 mg/kg, s.c.) were tested against the acetic acid-induced nociception, carrageenan-induced acute inflammatory paw edema/hyperalgesia, formalin-induced nociception and carrageenan-induced pleurisy in Swiss mice. RESULTS: Flavonoids rutin was detected in the phytochemical analysis of this extract. Oral treatment of AESM 300 mg/kg significantly reduced the number of acetic acid-induced abdominal writhing. AESM (100 and 300 mg/kg) significantly inhibited formalin-induced nociception, mechanical hyperalgesia and paw edema in carrageenan-model. Furthermore, AESM significantly inhibited leukocyte migration and protein exudation in the carrageenan-induced pleurisy test. CONCLUSION: This study confirms the antinociceptive, and anti-inflammatory activity of AESM, which may explain, in part, the popular use of this plant as a natural antinociceptive agent. This pharmacological action can be caused by flavonoids such as rutin and other compounds present in AESM.

Anti-hyperglycemic potential and chemical constituents of Aristolochia triangularis Cham. leaves - A medicinal species native to Brazilian forests.

ETHNOPHARMACOLOGICAL RELEVANCE: Aristolochia triangularis Cham. has been used in Brazilian traditional medicine for various therapeutic purposes, including as a leaf-based infusion for diabetes management. AIM OF THE STUDY: This study was designed to chemically characterize an infusion of in natura A. triangularis leaves and evaluate the in vivo anti-hyperglycemic properties of this infusion. MATERIALS AND METHODS: Chemical composition was examined using liquid-liquid extraction procedure, chromatographic methods, NMR, and LC-MS/MS. The in vivo anti-hyperglycemic activity of the freeze-dried infusion of A. triangularis leaves (Inf-L-At) was assessed using oral glucose tolerance test (OGTT). Initially, normoglycemic male rats were pre-treated with orally administered Inf-L-At at doses of 62.5, 125, and 250 mg/kg for two consecutive days. On the day of the OGTT, fasting animals received a glucose load (4 g/kg) 30 min after treatment with Inf-L-At, and the blood glucose levels were verified at 15, 30, 60, and 180 min. Intestinal maltase, lactase, and sucrase activities and muscle and liver glycogen contents were also assessed after the OGTT. RESULTS: Inf-L-At extract led to glycemic reduction with no dose-response at 15, 30, and 60 min comparable to that of the antidiabetic drug glibenclamide and was accompanied by an increase in hepatic and muscle glycogen contents. Additionally, there was a significant statistically decrease in the in vitro activity of disaccharidases. Maltase and sucrase activities were inhibited at all doses, whereas lactase activity was inhibited only at 62.5 and 250 mg/kg. In total, 75 compounds were found in the infusion, including seven new ones, (7S*,8S*,7ꞌS*,8ꞌR*)-4,4ꞌ-dihydroxy-3,3ꞌ-dimethoxy-7,9ꞌ-epoxylignan-7ꞌ-ol; 4ꞌ-hydroxy-3ꞌ-methoxy-3,4-methylenedioxy-7,9ꞌ-epoxylignan-9,7ꞌ-diol; triangularisines A, B, and C; N-ethyl-N-methyl-affineine; and N-methyl pachyconfine, and one previously not described as a natural product, epi-secoisolariciresinol monomethyl ether. CONCLUSION: The results demonstrated the anti-hyperglycemic activity of the infusion from A. triangularis leaves and showed that it is a rich source of lignoids, alkaloids, and glycosylated flavonoids, which are known to exhibit antidiabetic effects and other biological properties that can be beneficial for patients with chronic hyperglycemia, thus certifying the popular use of this herbal drink.

Selective anticancer effects of Serjania marginata Casar. extract in gastric cells are mediated by antioxidant response.

Gastric cancer is the fifth most common malignancy worldwide. Serjania marginata Casar. (SM) displays anti-inflammatory properties and has been used to treat gastrointestinal disorders. In the current study, we examined whether the hydroethanolic extract of SM leaves exerted cytotoxic, mutagenic, and protective effects in non-tumor gastric epithelium cells (MNP01) and gastric adenocarcinoma cells (ACP02) in vitro and analyzed whether its action was selective. Initially, cell viability (MTT assay), cell cycle kinetics (flow cytometry), and cell proliferation (total protein content) were analyzed. In addition, genomic instability (cytokinesis-block micronucleus cytome assay), anti/pro-oxidant status (CM-H2 DCFDA probe), and transcriptional expression (RT-qPCR) of genes related to cell cycle, cell death, and antioxidant defense were also evaluated. The SM extract was cytotoxic toward MNP01 and ACP02 cells at concentrations greater than 300 and 100 µg·ml-1 , respectively, and decreased protein content only toward ACP02 cells at 200 µg ml-1 . In ACP02 cells, the SM extract at 100 µg·ml-1 associated with doxorubicin (DXR; 0.2 µg ml-1 ) clearly promoted cell cycle arrest at the G2/M phase. The extract alone was not mutagenic to either cell type and reversed DXR-induced DNA damage and H2 O2 -induced oxidative stress in MNP01 cells. The gene expression experiments showed that SM hydroethanolic extract exerts an antioxidant response via NFE2L2 activation in non-tumor gastric cells, and cell cycle arrest (G2/M) in ACP02 gastric cancer cells via the TP53 pathway. The selective action of SM indicates that it is a promising therapeutic agent to treat gastric diseases and merits further studies.

Investigation of the antioxidant and hypoglycemiant properties of Alibertia edulis (L.C. Rich.) A.C. Rich. leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Alibertia edulis (L.C. Rich.) A.C. Rich is a vegetable species used in Brazilian folk medicine due to it is putative hypoglycemiant effect but has never been pharmacologically investigated. It is popularly used for the control of diabetes, especially in the state of Mato Grosso, Brazil. Following confirmation of the antioxidant activity of A. edulis by Aquino et al. (2017), the aim of this study was to evaluate the effects of leaves of A. edulis aqueous extract (AEAE) on some biochemical parameters in mice fed a high-fat fed. MATERIAL AND METHODS: Leaves of A. edulis were air-dried in an oven at 40 °C for 10 days and ground into a fine powder by mechanical milling. The AEAE was prepared by decoction (1:10 w/v) at 97 °C for 15 min, and later filtered and lyophilized. Preliminary phytochemical analysis of the AEAE has been already indetified the presence of caffeic acid, quercetin 3-rhamnosyl-(1 â†’ 6)-galactoside and iridois ioxide, ferulic acid and rutin in decocted leaves (Aquino et al., 2017). In one experiment, the acute oral toxicity AEAE was evaluated at 2,000 mg/kg of body weight. The animals were observed periodically for 14 days. In second experiment, the animals were divided into four groups (n = 5): Control, AEAE 200, AEAE 400 mg/kg and positive control (Metformin 100 mg/kg). In a third experiment, animals were divided into: Control RC (standard diet) (n = 24) and Control HFF (high-fat fed) (n = 24) groups for induction of glucose intolerance. After eight weeks, they were further subdivided into six groups (n = 8 each) RC or HFF with or without AEAE at doses of 200 and 400 mg/kg (2-wk) treatments to assess glucose tolerance. Plasma indicators of glucose tolerance and liver damage, skeletal muscle expression of antioxidant enzymes, and expression of the antioxidant proteins of superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and phosphorylated IKK were determined. RESULTS: The HF-fed animals developed glucose intolerance which the AEAE failed to revert. Meanwhile, the AEAE treatment did lower the glucose levels in the normolipidic cohorts by virtue of its antioxidant property. It was also observed that the treatment with the AEAE reduced food intake negatively interfering weight accretion. Beyond that, the treatment with AEAE interfered in the SOD and catalase expression and inhibited phosphorylation of IKK thus suggesting that the observed hypoglycemiant power may be related to its known antioxidant potential. No sings of toxicity or hemolysis were detectaed at indicating that, at the concentrations evaluated, the extract was not toxic to normal cells. CONCLUSION: The AEAE showed a hypoglycemiant effect in the normolipidic mice that received the control diet, but not in those that were made glucose-intolerant by consuming a high-fat fed. The extract also exhibited substantial protection against hemolysis and oxidative stress. Moreover, no signs of toxicity were evident at 2000 mg/kg of body weight.

Hepatic and gastroprotective activity of Serjania marginata leaf aqueous extract in Nile tilapia (Oreochromis niloticus).

The aim of this study was to assess the leaf aqueous extract composition of Serjania marginata and the effects of its inclusion on the diet of Nile tilapia (Oreochromis niloticus), with respect to the activity of digestive enzymes and enzymes associated with the metabolism of the liver and intestine and liver histopathology. Fish (initial mean weight, 54.36 ± 17.04 g) were divided into groups: fasting (without feeding), control (commercial feed), and treatment (commercial feed with leaf aqueous extract of Serjania marginata), and in each aquarium, there were five individuals and the fish fed ad libitum for 15 days. Treatment fish had ingested on average 224.3 mg of extract/kg of fish/day. In the extract analysis by mass spectrometry, quercitrin, isoquercitrin, A-type proanthocyanidin trimer, and quinic acid were identified. In the enzymatic activity, fish from the treatment group showed higher level of alkaline phosphatase, while the hepatotoxic markers (AST and ALT) and levels of lipase, amylase, and nonspecific protease did not differ (p > 0.05). In liver histopathological analysis, it was observed that fish from the treatment showed normal structure, while abnormalities were associated with control (fibrosis, loss of cordonal architecture, vacuolated hepatocytes with nucleus displaced to the periphery) and fasting (reduction in hepatocyte size and sinusoidal space). The intestine histopathology evidenced that the extract favored the development of goblet cells and intestinal fold height. The results indicated that the leaf aqueous extract of S. marginata assists in the structural maintenance of the liver and intestine and stimulates intestinal alkaline phosphatase production in Nile tilapia, suggesting that the identified compounds act on the liver and intestine, showing hepatoprotective effects and stimulating intestinal digestion.

Effects of exposure to ethanolic extract from Achyrocline satureioides (Lam.) D.C. flowers on reproductive and developmental parameters in Wistar rats.

Achyrocline satureioides (LAM) D.C. is a species plant used in folk medicine with several medicinal properties; however, few studies have focused on its potential adverse effects. The aim of this study was to examine the effects of ethanolic extract of A. satureioides flowers administered during pre-mating, mating, pregnancy and postpartum period on reproductive and developmental parameters in rats. Male and female rats received by gavage 0, 250, 500 or 750 mg/kg of extract. The animals were treated from pre-mating until 13 days post-partum. Phytochemical analysis revealed the presence of important flavonoids (quercetin, luteolin, caffeic acid, rutin, and ferulic acid). In females, biochemical, hematological or gestational parameters were not markedly altered by the extract. However, an increase in calcium and thyroid stimulating hormone (TSH) levels was found in treated-dams. Although TSH and T4 levels were not significantly altered in pups, there was a rise in body weight of pups whose mothers were treated with the extract. All males treated were able to successfully copulate with treated-females. However, rats exposed to 500 and 750 mg/kg of extract exhibited a significant decrease in daily testicular sperm production and delay in sperm transit time in the epididymis. The ethanolic extract of A. satureioides flowers produced adverse effects in the male reproductive system as evidenced by diminished sperm production and transport. In addition, the extract elevated TSH levels of exposed mothers which may consequently affect the development of pups but this requires further evaluation.

Toxicological safety evaluation in acute and 28-day studies of aqueous extract from Serjania marginata Casar. (Sapindaceae) leaves in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Serjania marginata Casar. (Sapindaceae) is a species traditionally known to be used for the treatment of gastric pain and inflammatory symptoms. AIM OF THIS STUDY: Due to the therapeutic importance of this species, this study investigated the toxicological effects of S. marginata leaves (AESM), after a single and a repeated exposure in rats. MATERIALS AND METHODS: For the acute toxicity test, 2000 mg/kg of AESM was administered to male and female rats by gavage, whereas for subacute toxicity test, 30, 150, or 750 mg/kg of AESM were used. RESULTS: No evidence of toxicity was observed in the animals acutely exposed to the extract, indicating that the LD50 is higher than 2000 mg/kg. After the repeated administration of AESM the hematological and biochemical parameters were unaltered, except the erythrocytes number and albumin levels in the exposed animals. Moreover, daily administration of this extract caused alteration on kidney histology. AESM also induced an increase of abnormal sperm, however the other reproductive parameters analyzed, in both sexes, were not altered by the treatment. CONCLUSION: Although AESM was not toxic after a single exposure, its use after prolonged periods affected some analyzed parameters, indicating that precautions should be taken when it is given over longer periods.

Evaluation of the toxicity and anti-inflammatory activities of the infusion of leaves of Campomanesia guazumifolia (Cambess.) O. Berg.

ETHNOPHARMACOLOGICAL RELEVANCE: Some species of Campomanesia are used in the folk medicine due to anti-inflammatory, anti-diarrheal, anti-diabetes and hypercholesterolemic. However studies with Campomanesia guazumifolia (Cambess.) O. Berg. are scarce. AIM OF THE STUDY: This study investigated the anti-inflammatory activity and toxicological profile of infusion obtained from leaves of Campomanesia guazumifolia in mice. MATERIALS AND METHODS: Leaves infusion of C. guazumifolia was obtained in the proportion of 20 g/L (leaves/water) at 95-100 °C for 10 min in an enclosed container. The acute toxicity of the leaves infusion of C. guazumifolia lyophilized (ICG) was assessed by oral administration to female mice at doses of 500, 1000, 2000, and 5000 mg/kg, and the general behavior and toxic symptoms were observed for 14 days. In the subacute toxicity model, female mice were treated orally with the ICG (250, 500, and 1000 mg/kg) during 28 days, and biochemical, toxic signs and the estrous cycle were evaluated. The anti-inflammatory activity of the ICG (70, 300 and 700 mg/kg) was analyzed using carrageenan-induced pleurisy and inflammatory paw (mechanical and thermal hyperalgesia). RESULTS: Three flavonoids glycosylated and a cyclohexanecarboxylic acid were identified in the ICG: quercetin pentose, quercetin deoxyhexoside, myricetin deoxyhexoside and quinic acid. No clinical signs of acute toxicity were observed, suggesting that the LD50 (Lethal Dose) is above 5000 mg/kg. Subacute exposure of mice to the ICG did not change significantly the hematological and biochemical parameters as well as histology of organs. The ICG increased the duration of estrous cycle in all phases, showing anti-inflammatory potential by decreasing leukocyte migration, extravasation protein in the pleural cavity and antiedematogenic activity. The ICG treatment at a dose of 700 mg/kg decreased the mechanical hyperalgesia, while at doses of 300 mg/kg and 700 mg/kg, decreased the sensitivity to the cold. CONCLUSION: The results evidenced the anti-inflammatory potential with low toxicity of infusion of the leaves of C. guazumifolia, supporting the popular use of this species.

Alibertia edulis (L.C. Rich.) A.C. Rich - A potent diuretic arising from Brazilian indigenous species.

ETHNOPHARMACOLOGICAL RELEVANCE: Although Alibertia edulis (L.C. Rich.) A.C. Rich decoction is used in Brazilian folk medicine due to its possible antihypertensive effect, this species has never been critically investigated as a hypotensive drug. So, the aim of this study was to evaluate the possible hypotensive and antihypertensive effects of the oral administration of Alibertia edulis aqueous extract (AEAE) in normotensive and hypertensive rats, and evaluate its inter-relation with a possible diuretic activity. MATERIAL AND METHODS: Different doses of AEAE (20, 65 and 200mg/kg) were tested on the mean arterial pressure (MAP) of normotensive Wistar rats and after induction of renovascular hypertension (two-kidney, one-clip Goldblatt model). In addition, the diuretic effects of AEAE were compared with hydrochlorothiazide (HCTZ) in an acute and repeated-dose treatment for 7 days. Volume, sodium, potassium, chloride, calcium contents, pH and density were estimated in urine samples collected after 8 or 24h. Plasma sodium, potassium, total protein, urea, creatinine, AST and ALT concentrations were measured in samples collected at the end of the experimental period (seventh day). Finally, the antioxidant activity of the AEAE was assessed using the DPPH radical scavenging and ferric ions reducing power assay. RESULTS: The intraduodenal administration of the HCTZ and AEAE significantly reduced, in a dose-dependent manner, the MAP in both normotensive and hypertensive rats. Otherwise, the heart rate was not affected by any treatment. Acute and prolonged oral administration of AEAE (200mg/kg) and HCTZ caused a significant increase in volume and urinary concentrations of sodium, potassium and chloride. Moreover, urinary calcium concentration was significantly increased after administration of AEAE (200mg/kg). Finally, AEAE was able to present important in vitro antioxidant properties. CONCLUSION: The results obtained have shown that AEAE presents potent diuretic activity and significant hypotensive and antihypertensive effect. In addition, this study may confirm part of the pharmacological activity popularly attributed to this species and opens perspective for the future use in various renal and cardiovascular diseases.

Acute and subacute toxicity of the aqueous extract of Alibertia edulis (Rich.) A. Rich. ex DC. in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Alibertia edulis, popularly known as "marmelo do Cerrado" is a native plant from the brazilian Cerrado. It has high food and ornamental potential and the tea leaves are currently used as hypoglycemic, antihypertensive and diuretic. AIM OF THE STUDY: In order to evaluate the safety of the aqueous extract of Alibertia edulis leaves (AEAE), the acute and subacute toxicity tests were performed in male and female Wistar albino rats. MATERIALS AND METHODS: The experiments were performed in accordance with the OECD guidelines 425 and 407. For the acute toxicity, one single dose of the AEAE (2000mg/kg) was administered by gavage to five female rats. The animals were observed for 14 days for any signs of toxicity and death. In the subacute toxicity, four different doses (125, 250, 500 and 1000mg/kg) of the AEAE were administered to male and female rats for 28 consecutive days. A satellite group received the maximum dose (1000mg/kg) for 28 days and remained untreated for 14 more days in order to observe reversibility, persistence, or delayed occurrence of toxic effects. The five parameters of the Hippocratic screening, body weight, food and water intake were daily observed. At the end of the experiment, blood samples were collected for the hematological and biochemical analysis. The vital and reproductive organs were inspected for any histopathological changes. RESULTS: No deaths or behavioral changes were observed during both experiments as well as no changes on organ weights, biochemical, hematological and histopathological parameters that could indicate any signs of toxicity. CONCLUSION: All doses tested can be considered safe in rats and the LD50 is higher than 2000mg/kg. Therefore, further assessments are required in order to proceed to clinical studies in humans.

Study on the Cytotoxic, Genotoxic and Clastogenic Potential of Attalea phalerata Mart. ex Spreng. Oil Pulp In Vitro and In Vivo Experimental Models.

Attalea phalerata Mart. ex Spreng. (Arecaceae), popularly known as "bacuri", is used in Brazilian folk medicine. Its oil is used orally to relieve pulmonary congestion and joint pain. In topical applications, it is applied as an effective hair tonic and anti-dandruff. The in natura pulp and its nuts are used as food because of its nutritional value. Despite its use in folk medicine, there is a lack of data regarding its in vivo/in vitro cytotoxic/genotoxic and clastogenic effects. Therefore, in this study, we evaluated the cytotoxic, genotoxic and clastogenic effects of Attalea phalerata Mart. ex Spreng. oil (APMO) in vitro and in vivo. For the analysis of cytotoxic potential, the Artemia salina and MTT (3-(4,5-dimethizzol-zyl)-2,5-diphenyltetrazolium bromide) assays were performed. Possible cytotoxic, genotoxic and clastogenic effects of APMO intake were determined by performing the comet and micronucleus assays. Male and female Wistar rats were orally treated with doses of 125, 250, 500 or 1000 mg.kg-1 of the APMO daily for 28 consecutive days (four weeks). The results showed that the APMO did not induce cell death in the experiments of Artemia salina and MTT, indicating that it has no cytotoxicity. The APMO did not cause significant damage to the DNA of the rats in the four doses used when compared to the negative control group (saline + Tween® 80). The APMO did not present any significant increase in micronucleated polychromatic erythrocytes (MNPCEs) for the four tested doses. When compared to the positive control group, all groups (comet and micronucleus tests) were statistically different. These data suggest that the administration of Attalea phalerata Mart oil. ex Spreng does not cause cytotoxicity, genotoxicity and clastogenicity in experimental models in vitro and in vivo following oral administration in this study.

Does the gastroprotective action of a medicinal plant ensure healing effects? An integrative study of the biological effects of Serjania marginata Casar. (Sapindaceae) in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Serjania marginata (Sapindaceae), a medicinal plant commonly found in the Brazilian Cerrado, Paraguay, Bolivia and Argentina, is also known as "cipó-uva" or "cipó-timbó". Ethnopharmacological studies indicate that the leaves from this medicinal plant are used in folk medicine to treat gastric pain. The overall objective of this study was to evaluate the gastroprotective and healing effect of the hydroalcoholic extract obtained from S. marginata (HESM) leaves using rodent experimental models. As part of the integrative study of this medicinal plant, we also evaluated the acute toxicity, antimicrobial, antidiarrheal, (anti)mutagenic, and hemodynamic effects. MATERIAL AND METHODS: We performed a pharmacological study to test the acute toxicity and antimutagenic effect (Ames assay) of the HESM. The HESM was tested against different necrosis-promoting agents and experimental manipulations, such as absolute ethanol, cysteamine, pyloric ligature, and ischemia-reperfusion (I/R) injury. The gastroprotective effect of the HESM was assessed by analyzing the gastric juice (volume, pH, total acidity) and the mucus in the gastric mucosa from rats. We assessed the levels of NO, sulfhydryl compounds, PGE2, vanilloid receptor, glutathione (GSH), and malondialdehyde (MDA), as well as the myeloperoxidase (MPO) activity. The gastric healing effects of the HESM were evaluated during 7 or 14 days of treatment. The intestinal motility, antidiarrheal action, and antibacterial effects (microdilution methods) of the HESM were also evaluated. RESULTS: The phytochemical analysis of the HESM revealed the presence of saponins, flavonoid glycosides, and tannins. The extract exhibited no sign of acute toxicity or mutagenic effect in vitro. In contrast, this extract exhibited a protective effect against the mutagenic action of direct- and indirect-acting mutagens. Only the oral administration of HESM (250mg/kg) significantly decreased the severity of gastric damage induced by ethanol (60.13%) and I/R (58.31%). The HESM exerts its gastroprotective effects by decreasing the MPO and MDA activities in the gastric tissue and by increasing the amount of adherent mucus covering the gastric mucosa. In vitro, the extract also displayed evident antimicrobial effects against Helicobacter pylori. However, the preventive effect of the HESM was not accompanied by an ulcer-healing effect. The treatment with HESM (14 days) significantly increased gastric lesions in 99% of the tested animals compared with the control group. This result represents a highly relevant piece of evidence that should resonate as an alert against the chronic use of this medicinal plant as an antiulcer in folk medicine. CONCLUSIONS: Despite the anti-H. pylori and gastroprotective actions of S. marginata in experimental models, the gastric injuries aggravation induced after chronic treatment with the HESM argues against the use of this plant species in folk medicine.