Hancornia speciosa: indications of gastroprotective, healing and anti-Helicobacter pylori actions

Ethnopharmacological relevance

Mangaba (Hancornia speciosa Gomez) is a medicinal plant frequently cited in ethnopharmacological inventories of the central region of Brazil against gastrointestinal disorders such as diarrhoea, ulcer, gastritis and stomachache.

Aim of the study

The hydroalcoholic extract (HE) and infusion (BI) of Hancornia speciosa bark were investigated for their ability to prevent and heal rodent gastric ulcer.

Materials and methods

The preventive and healing action of both preparations of Hancornia speciosa were evaluated in experimental models in rodents that simulated this disease in human gastric mucosa.

Results

BI did not exert gastroprotective effect, in contrast to HE (500 mg/kg, p.o.) that decreased (p < 0.05) the severity of gastric damage induced by HCl/ethanol (52%), indomethacin/bethanechol (51%), stress (52%) or pylorus ligature experiments (54%). HE increased (p < 0.05) the pH and decreased acid output of gastric juice. This extract promoted increase of mucus amount (3.62 mg/wt. tissue vs. 5.81 mg/wt. tissue), healing action (67%) and displayed anti-Helicobacter pylori effect.

Conclusions

The antiulcer action of Hancornia speciosa resulted in increase of gastric mucus formation and antioxidant properties of polymeric proanthocyanidins present in the bark composition of this medicinal plant.     

Antiulcerogenic activity of hydroalcoholic fruit extract of Pithecellobium dulce in different experimental ulcer models in rats

Ethnopharmacological relevance

The ethnopharmacological importance of Pithecellobium dulce is evidenced by its traditional use for gastric complications. The aim of the study is to evaluate the gastroprotective activity and the mechanism of action of hydroalcoholic fruit extract of P. dulce (HAEPD) in rats by using chemical and stress induced ulcer models.

Materials and methods

Gastric ulcer was induced by administering alcohol (or) acetylsalicylic acid (or) hypothermic restraint stress to rats pretreated with HAEPD (200 mg/kg b wt for 30 day). Volume of gastric fluid, pH, acidity, activities of pepsin, H⁺, K⁺-ATPase, myeloperoxidase, mucin content, nucleic acids, glycoproteins and prostaglandin E₂ (PGE₂) levels were assessed in gastric tissues.

Results

Ulcer score was significantly minimized in HAEPD administered animals. pH and acidity of gastric fluid were significantly minimized and the mucin, PGE₂ levels were significantly maintained in drug pre administered animals. The activities of H⁺, K⁺- ATPase and myeloperoxidase were found to be significantly elevated in ulcer control animals and found to be decreased in drug pretreated animals. The cell proliferation was found to be enhanced in drug received animals. The total protein bound carbohydrate to total protein ratio was found to be significantly maintained by HAEPD. The effects were found to be comparable with that of standard drug omeprazole.

Conclusion

It is concluded that HAEPD possess a potent antiulcer activity probably by acting as cytoprotective and antiacid secretory agent.     

Antibacterial activity of the phenolic acids fractions of Scrophularia frutescens and Scrophularia sambucifolia

The phenolic fractions of aerial part of Scrophularia frutescens and S. sambucifolia (Scrophulariaceae) showed a potent antibacterial activity. Ferulic, isovanillic, p-hydroxycinnamic, p-hydroxybenzoic, syringic, caffeic, gentisic and protocatechuic acids were isolated from S. frutescens and ferulic, p-coumaric, vanillic, p-hydroxibenzoic and syringic acids were isolated from S. sambucifolia. Since phenolic acids have been shown in the literature to exert an antibacterial effect, the presence of these compounds in the two plants explains their antibacterial activity.     

Gastroprotective activity of hydroalcoholic extract obtained from the leaves of Brassica oleracea var. acephala DC in different animal models

Ethnopharmacological relevance

Brassica oleracea var. acephala DC has been extensively used in Brazilian traditional medicine to treat gastric ulcer.

Aim of the study

This study was conducted to evaluate the antiulcerogenic property of hydroalcoholic extract obtained from the leaves of Brassica oleracea.

Materials and methods

Antiulcer assays were performed using the protocol of ulcer induced by ethanol/HCl, and non-steroidal anti-inflammatory drugs (NSAIDs). Parameters of gastric secretion (volume, pH and [H⁺]) were determined by the pylorus ligation model and mucus in gastric contents.

Results

In the ethanol-induced ulcer model, we observed a significant reduction in all the parameters analyzed, obtaining curative ratios of 58.8 ± 11.5, 86.2 ± 12.2 and 42.8 ± 6.6% for the groups treated with 50 and 100 mg/kg of extract and omeprazole (30 mg/kg), respectively. The dose of 25 mg/kg of hydroalcoholic extract of Brassica oleracea showed no significant results. In the indomethacin-induced ulcer, the percentages of ulcer inhibition were 64.3 ± 9.9, 66.4 ± 12.3 and 81.2 ± 7.5% for the groups treated with 50 and 100 mg/kg extract and positive control (cimetidine, 100 mg/kg), respectively. The results showed a significant increase in pH and mucus production in the groups treated with Brassica oleracea when compared with the control group. No sign of toxicity was observed in the acute toxicity study.

Conclusions

The results of the present study show that hydroalcoholic extract of Brassica oleracea displays antiulcer activity, as demonstrated by the significant inhibition of ulcer formation induced using different models. The data suggest that the effectiveness of the extract is based on its ability to stimulate the synthesis of mucus, increase pH and decrease H⁺ ions in the stomach. This work corroborates the ethnopharmacology use of Brassica oleracea preparations, contributing to its pharmacological validation by suggesting that preparations obtained from Brassica oleracea could be used for the development of new phytopharmaceuticals for the treatment of gastric ulcer.     

Qingkailing injection attenuates apoptosis and neurologic deficits in a rat model of intracerebral hemorrhage

Aim of the study

Traditional Chinese herb Angong Niuhuang Pill (AGNHP) is a famous preparation for neurological diseases; Qingkailing injection (QKL), an extract of AGNHP has similar clinical applications. This investigation was designed to further elucidate the neuroprotective effect of QKL on intracerebral hemorrhage (ICH).

Materials and methods

ICH was produced in adult Sprague–Dawley rats by injection of collagenase IV. Three incremental doses of QKL injection including low-(0.5 ml/kg), moderate-(1 ml/kg) and high-dosage (2 ml/kg) were administered twice, 3 and 12 h following ICH. TUNEL and caspase-3 activity were measured at 1 d after ICH, and apomorphine-induced rotation was evaluated at 1 d, 7 d, 14 d and 28 d.

Results

Administration of high-dose QKL inhibited TUNEL positive cells (p < 0.05) and caspase-3 activity (p < 0.05) at 1 d following ICH, and reduced apomorphine-induced rotation at 1 d (p < 0.01), 7 d, 14 d and 28 d (p < 0.05), compared with the controls. However, QKL in a low or moderate dose had no such effect.

Conclusion

QKL reduced brain damage of intracerebral hemorrhage through inhibiting apoptosis, which suggested a potential intervention for ICH patients.     

Evaluation of antiulcer activity and mechanism of action of methanol stem bark extract of Lafoensia pacari A. St.-Hil. (Lytraceae) in experimental animals

Ethnopharmacological relevance

Lafoensia pacari St.-Hil. is a tree native to the Brazilian Cerrado. Its bark macerate is popularly used in Brazilian folk medicine for the treatment of peptic ulcer, wound healing and inflammation.

Aim of the study

To evaluate the antiulcer activities of the methanol extract of Lafoensia pacari (MELP) and possible mechanisms of actions involved.

Materials and methods

The stem bark of Lafoensia pacari was macerated in methanol to obtain the crude methanol extract of Lafoensia pacari (MELP). The gastroprotective and ulcer healing of MELP were evaluated using ethanol, indomethacin, cold-restraint stress-induced (acute) and acetic acid (chronic) ulcer models. The probable mode of action of MELP was also evaluated by determining intestinal transit, involvement of non-protein sulfhyhydryls (NP-SH), gastric mucus secretion, gastric secretory parameters and pro-inflammatory cytokines, tumor necrosis factor-α (TNF-α) and interleukin-17 (IL-17). Phytochemical analysis was carried out using classical methods and ellagic acid, one of the most important pharmacological active compounds was confirmed by HPLC.

Results

The results of the pharmacological studies on MELP demonstrated potent and effective gastroprotection against ethanol, indomethacin and cold stress-induced acute ulcers and ulcer healing in acetic acid induced chronic ulcer. MELP had no significant effect on the intestinal motility and it is also independent of mucus production but rather have a mucolytic effect. In pylorus-ligated rats the extract showed anti-secretory activity by decreasing total gastric juice volume and gastric acidity while increasing the gastric pH. The gastroprotection against ethanol is partially attributed to effective attenuation in the decrease in NP-SH levels, inhibition of the increases in the pro-inflammatory cytokines, TNF-α and IL-17. Phytochemical analysis of MELP revealed the presence of pyrrogalic tannins, saponins, steroids, triterpenoids and simple phenols, with ellagic acid being the major components.

Conclusion

The results of this study showed that MELP possesses preventive and curative effects against gastric ulcer in experimental animals. These effects are partly dependent on its anti-oxidant, anti-secretory properties and inhibition of pro-inflammatory cytokines and independent of gastric/intestinal motility and mucus secretion. Ellagic acid, a compound of proven antiulcer activity, was found in MELP as the major component. These results confirmed the traditional use of MELP in Brazilian popular medicine for the treatment of gastric ulcers and shed some light on some of its mechanisms of action.     

HILIC quantification of oenotheralanosterol A and B from Oenothera biennis and their suppression of IL-6 and TNF-α expression in mouse macrophages

Ethnopharmacological relevance

Evening primrose (Oenothera biennis L.) is a wild medicinal herb of Central American origin that is now globally widespread. Its traditional uses include treatment of rheumatoid arthritis and premenopausal pain both of which have an inflammatory component. The present study demonstrates the in vitro anti-inflammatory activity of three Oenothera biennis compounds.

Materials and methods

Oenotheralanosterol A and B (Oen-A &; Oen-B) along with gallic acid (GA) were isolated and characterized using column chromatography and NMR. The compounds were tested with LPS stimulated peritoneal mouse macrophages assaying for suppression of IL-6, TNF-α and NO synthesis. An HILIC method for the simultaneous quantitation of GA, Oen-A, and Oen-B in Oenothera biennis plant material was also developed as a means of monitoring quality of plant material.

Results

Significant inhibition of TNF-α and IL-6 by GA, Oen-A and Oen-B was observed (p < 0.05). Inhibition was concentration dependent and no synergistic or antagonistic effect on pro-inflammatory cytokines was found when used in combination (1:1) (p > 0.05). The HILIC analysis method was validated using Oenothera biennis root.

Conclusion

The study demonstrates the anti-inflammatory activity of Oenothera biennis root compounds and supports its traditional use in arthritis management. Active anti-inflammatory compounds were identified and quantified by the HILIC method.     

Cerebral protection of purified Herba Leonuri extract on middle cerebral artery occluded rats

Aim of study

Oxidative stress is involved in stroke. In particular, Chinese Herbal Medicine with antioxidant properties is believed to have potential therapeutic effect. In this study, neuroprotective effects of purified Herba Leonuri (pHL) were evaluated in Wistar rats undergone middle cerebral artery occlusion (MCAO).

Materials and methods

The rats were treated with their respective treatments for 2 weeks prior to the MCAO, continually treated for another 7 days after MCAO. During the post-surgery treatment period, neurological deficit score was measured. At the end of treatment, animals were sacrificed and samples were collected for analysis of infarct volume, apoptosis and antioxidant analysis.

Results

Under the treatment of pHL, the infarct volume was reduced significantly from 20.75 ± 0.03% to 15.19 ± 0.02% (p < 0.05). The neurological impairment was alleviated to 1.82 as compared to vehicle (2.43). Plasma antioxidant concentration was increased from 0.31 ± 0.03 mM to 0.42 ± 0.05 mM (p < 0.05). DNA oxidative damage was reduced to 1.19 ± 0.03 in stroke pHL treated group (p < 0.05 as compared to vehicle group, 1.78 ± 0.03). pHL could reduce the level of apoptosis and also the pro-apoptotic proteins, but increase the level of anti-apoptotic proteins.

Conclusion

pHL is believed to have promising therapeutic effect for stroke treatment through antioxidant mechanisms.     

Evaluation of antiulcer activity of chromanone fraction from Calophyllum brasiliesnse Camb

Ethnopharmacological relevance

Calophyllum brasiliense Camb. (Clusiaceae), popularly known as ‘guanandi’, is found in the tropical areas and swampy lands. The latex exuding from its bark is used in the treatment of gastric ulcer in folk medicine. Several active compounds have been isolated from its stem bark among them, are the chromanone acids. Therefore, this study aimed to evaluate antiulcer activity and probable mechanism(s) of action of a fraction containing a mixture of chromanone acids (BI), derived by column chromatography fractionation of the hexane extract of the stem bark of Calophyllum brasiliense (HECb), using experimental in vitro and in vivo models.

Materials and methods

Ulcer was induced by oral administration of ethanol (75%, v/v) and indomethacin (50 mg/kg). Malondialdehyde, reduced glutathione and catalase activity was measured in stomach tissue after ethanol induced ulcer. In order to evaluate the effect of BI on nitric oxide, ulcer was induced by ethanol in l-NAME pretreated animals. Anti-Helicobacter pylori activity was verified in disk diffusion and broth microdilution in vitro assays, using cagA+ and vacA+ Helicobacter pylori strains.

Results

BI prevented the gastric ulceration caused by ethanol and indomethacin treatments. Its gastroprotective mechanism in ethanol-induced ulcer was partly due to reduction of MDA and CAT levels in the gastric tissue. BI did not affect the GSH levels and its gastroprotective effect was not reversed by pretreatment with l-NAME. BI showed anti-Helicobacter pylori in the both assays.

Conclusion

The results indicate that BI is partly responsible for the HECb antiulcer and anti-Helicobacter pylori effects.     

Gastroprotective activities of Turnera diffusa Willd. ex Schult. revisited: Role of arbutin

Ethnopharmacological relevance

Turnera diffusa Willd. ex Schult. has been used for the treatment of several human disorders including peptic ulcer.

Objectives of the study

The current study is an attempt to evaluate the anti-ulcerogenic activities of arbutin, a major constituent of Turnera diffusa on two ulcer models. The possible involvement of lipid peroxidation, nitric oxide, IL-6, IL-10, TNF-α and mucus barrier mechanism has been investigated.

Materials and methods

Effects of arbutin on ulcer index, gastric juice acidity, mucus content and histochemistry, gross and histological gastric lesions, nitric oxide, cytokines levels (IL-6, IL-10 and TNF-α), and thiobarbituric acid reactive substances (TBARS), were evaluated in aspirin or ethanol-induced ulcer in vivo. Acute toxicity of arbutin was also examined in rodent model. MTT assay was used to assess the cytotoxicity of the compound on normal liver cells (WRL-68).

Results

Pre-treatment with arbutin or omeprazole protected the gastric mucosa as seen by reduction in ulcer area and mucosal content, reduced or absence of edema, inflammation and leucocytes infiltration on both models. Arbutin significantly (P < 0.05) lowered the elevated TBARS level into gasteric homogenate. Arbutin did not produce significant inhibition of NO. This natural compound has modulated the levels of interleukin-6, interleukin-10 and TNF-α. No in vitro or in vivo toxicities for arbutin were observed.

Conclusion

Thus it can be concluded that Turnera diffusa possesses anti-ulcer activity, which could be attributed to lipid peroxidation inhibitory, immuno modulatory and anti-oxidant mechanisms of arbutin but not to the intervention with nitric oxide inflammation pathway.     

Effects of acute administration of the hydroalcoholic extract of mate tea leaves (Ilex paraguariensis) in animal models of learning and memory

Aim of the study

Ilex paraguariensis St. Hilaire (Aquifoliaceae) is a plant widely cultivated in South America that is used to prepare a tea-like beverage with a reputation to improve cognitive function, a response that has been attributed to the constituents of the leaves, especially caffeine. Our previous study indicated that the hydroalcoholic extract of Ilex paraguariensis presents an antiparkinsonian profile in reserpine- and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP)-treated rodents.

Materials and methods

In the present study, the effects of the hydroalcoholic extract of Ilex paraguariensis on the short- and long-term learning and memory of rats were assessed with the social recognition, Morris water maze, and step-down inhibitory avoidance tasks.

Results

A preliminary HPLC fingerprint of the plant extract confirmed the presence of caffeine (the major compound), rutin and kaemperol, and revealed the absence of detectable concentrations of caffeic acid, quercetin and ursolic acid. Acute pre-training intraperitoneal (i.p.) or oral administration of the extract of Ilex paraguariensis improved the short-term social memory in a specific manner as well as facilitated the step-down inhibitory avoidance short-term memory evaluated 1.5 h after training. Moreover, a synergistic response was observed following the co-administration of ‘non-effective’ doses of caffeine and Ilex paraguariensis in the social memory. In contrast, pre-training administration of hydroalcoholic extract of Ilex paraguariensis did not alter the step-down inhibitory avoidance long-term memory evaluated 24 h after training, while the highest dose tested (250 mg/kg, i.p.) disrupted the animals’ performance in a cued version of the Morris water maze.

Conclusion

These results partly substantiate the traditional use of mate tea for improvement of cognition indicating that acute administration of hydroalcoholic extract of Ilex paraguariensis differentially modulates short- and long-term learning and memory in rats probably through its antagonist's action on adenosine receptors.     

Antiulcer principle from Zingiber montanum

Ethnopharmacological relevance

Rhizome of Zingiber montanum has been extensively used as a folk medicine to ameliorate peptic ulcer at northern part of Bangladesh.

Aim of the study

To identify the antiulcer principle of the MeOH extract of the rhizome of Zingiber montanum by an ex vivo bioassay guided chromatographic separation and purification, and structure elucidation of the purified compound by spectroscopic methods.

Materials and methods

Dried powder of Zingiber montanum rhizomes was extracted with MeOH. The antiulcer activity of the crude extract and its chromatographic fractions were evaluated by the inhibition of 1 N HCl induced gastric lesions in Swiss albino mice. The pure compound was purified from the active fraction by crystallization with hexanes. Structure of the pure compound was elucidated by spectroscopic methods. The antiulcer activity of the pure compound was evaluated by the inhibition of 1 N HCl, 95% ethanol and indomethacin induced gastric lesions in mice.

Results

The MeOH extract of Zingiber montanum showed 61.97% and 83.10% inhibition of the 1 N HCl induced gastric lesions at doses of 200 mg/kg and 400 mg/kg, respectively, in mice. Chromatographic separation on silica gel of the extract was yielded seven fractions and the fraction 2 was found to have most potent antiulcer activity in mice. This fraction showed 77.46% inhibition of the 1 N HCl induced gastric lesions at a dose of 40 mg/kg in mice. Crystallization of the fraction yielded 1 (zerumbone, 180 mg). It showed statistically 45.77% and 92.25% inhibition of 1 N HCl induced gastric lesions in mice at doses of 20 mg/kg and 40 mg/kg, respectively. It also showed 29.07% and 45.35% inhibition of 95% ethanol induced gastric mucosal damage, and 64.76% and 72.38% inhibition of indomethacin induced gastric lesions in mice at doses of 20 mg/kg and 40 mg/kg, respectively.

Conclusion

Zerumbone (1) showed potent cytoprotective effect against necrotizing agent (HCl) and non-steroidal anti-inflammatory drug (indomethacin) induced gastric ulceration. It also exhibited moderate cytoprotective effect against noxious agent (EtOH) induced gastric lesions. It can be considered as a promising new antiulcer natural drug lead.     

Endothelium-independent relaxation and contraction of rat aorta induced by ethyl acetate extract from leaves of Morus alba (L.)

Aim of the study

Based on screening for vasoactive traditional Chinese medicinal herbs, the present study was performed to investigate the vasoactive effects of an ethyl acetate extract from leaves of Morus alba (L.) (ELM) on rat thoracic aorta and the mechanisms underlying these effects.

Materials and methods

Isolated rat thoracic rings were mounted in an organ bath system and the effects of ELM on their responses were evaluated.

Results

ELM (0.125–32.000 g/l) induced a concentration-dependent relaxation (P < 0.01 vs. control) both in endothelium-intact and -denuded aortas precontracted by high K⁺ (6 × 10⁻² M) or 10⁻⁶ M phenylephrine (PE). In endothelium-denuded aortas, ELM at the EC₅₀ concentration reduced Ca²⁺-induced contraction (P < 0.01 vs. control) after PE or KCl had generated a stable contraction in Ca²⁺-free solution. And after incubation with verapamil, ELM induced contraction in endothelium-denuded aortas precontracted by PE (P < 0.01 vs. control); this was abolished by ruthenium red (P < 0.01 vs. ELM-treated endothelium-denuded group; P > 0.05 vs. control), but not by heparin (P > 0.01 vs. ELM-treated endothelium-denuded group; P < 0.01 vs. control).

Conclusions

The results showed that ELM had dual vasoactive effects, and the relaxation was greater than the contraction. The relaxation was mediated by inhibition of voltage- and receptor-dependent Ca²⁺ channels in vascular smooth muscle cells, while the contraction occurred via activation of ryanodine receptors in the sarcoplasmic reticulum.     

Evaluation of the genotoxic and antigenotoxic potential of Brassica oleracea L. var. acephala D.C. in different cells of mice

Ethnopharmacological relevance

Brassica oleracea L. var. acephala D.C. has been extensively used in Brazilian traditional medicine to treat gastric ulcer.

Aim of the study

This study was conducted to evaluate the in vivo genotoxic and/or antigenotoxic potential of a Brassica oleraceae hydroalcoholic extract obtained from the leaves, in different cells of mice.

Materials and methods

Analyses were performed using the comet assay, on leukocytes (collected 4 and 24 h after treatment), liver, brain, bone marrow and testicular cells (collected 24 h after treatment), and using the micronucleus test (MN) in bone marrow cells. Eight groups of albino Swiss mice were treated (N=6): control (C), positive control (doxorubicin 80 mg/kg (DXR)), and six experimental groups, which received 500, 1000 and 2000 mg/kg of Brassica oleraceae extract alone by gavage, while a further three groups received the same doses plus DXR (80 mg/kg). We calculated the damage scores, and their averages were compared by ANOVA followed by the Tukey test for multiple comparisons.

Results

The results demonstrated that none of the tested doses of Brassica oleraceae extract showed genotoxic effects by the comet assay, or clastogenic effects by the MN test. On the other hand, for all cells evaluated, the three tested doses of the Brassica extract promoted inhibition of DNA damage induced by DXR.

Conclusions

Under our experimental conditions, Brassica oleraceae leaf extract showed no genotoxic or clastogenic effects in different cells of mice. However, it did show a significant decrease in DNA damage induced by doxorubicin. It is suggested that the antigenotoxic properties of this extract may be of great pharmacological importance, and may be beneficial for cancer prevention.     

Inhibition of the human neutrophil oxidative metabolism by Baccharis dracunculifolia DC (Asteraceae) is influenced by seasonality and the ratio of caffeic acid to other phenolic compounds

Ethnopharmacological relevance

The great potential of phytotherapic drugs for treating and preventing inflammatory diseases mediated by increased neutrophil reactive oxygen species (ROS) generation has guided the search for new natural products with antioxidant and immunomodulatory properties. Baccharis dracunculifolia D.C. (Asteraceae), the main botanical source of Brazilian green propolis, is a native plant from Brazil widely used in folk medicine as anti-inflammatory. This study aims: (a) to determine the influence of seasonality on the chemical profile and biological activity of Baccharis dracunculifolia (Asteraceae) leaf extracts (BdE); (b) to analyze the correlation between the major compounds and the ability of BdE to modulate the superoxide anion and total ROS generation by human neutrophils.

Materials and methods

The extracts were obtained from leaf samples collected monthly during one year. The superoxide anion and total ROS generation were assessed by the lucigenin (CL-luc)- and luminol (CL-lum)-enhanced chemiluminescence assays.

Results

Seasonality influenced more the quantitative than the qualitative chemical profile of B. dracunculifolia, and affected its biological activity. The major compounds identified were caffeic acid, p-coumaric acid, aromadendrin-4′-methyl ether (AME), isosakuranetin and artepillin C. The IC₅₀ values obtained for CL-lum and CL-luc inhibition by BdE ranged from 8.1–15.8 and 5.8–13.3 µg mL⁻¹, respectively, and correlated positively with caffeic acid concentration. CL-luc inhibition correlated negatively with the concentration of artepillin C, AME, isosakuranetin and total flavonoids. The BdE sample from May/07 inhibited CL-lum and CL-luc the most strongly (IC₅₀=8.1±1.6 and 5.8±1.0 µg mL⁻¹, respectively), and contained the highest ratio of caffeic acid to the other isolated compounds; so, this ratio could be employed as chemical marker for this biological activity of B. dracunculifolia.

Conclusion

The ability of B. dracunculifolia to inhibit the neutrophil ROS generation depends more on the type and ratio of phenolic compounds and flavonoids than on their high absolute concentrations. Together, our results help select the most appropriate plant material for the production of phytotherapic drugs to be used in the treatment of inflammatory diseases mediated by increased neutrophil activation.     

Effect of Brazilian copaiba oils on Leishmania amazonensis

Ethnopharmacological relevance

Copaiba oil has been used in folk medicine since the 19th century. The use of copaiba oils to treat leishmaniasis is cited in several ethnopharmacological studies. Nevertheless, the potential antileishmania of copaiba oils had not been studied.

Aim of the study

Eight different kinds of Brazilian copaiba oils were screened for antileishmanial activity.

Materials and methods

The antiproliferative effect of copaiba oil on promastigote and amastigote axenic were determined. To determine the survival index peritoneal macrophage were infected with promastigotes of Leishmania amazonensis and treated with copaiba oil. The cytotoxic effect of copaiba oil was assessed on macrophage strain J774G8 by assay of sulforhodamine B.

Results

Copaiba oils showed variable levels of activity against promastigote forms with IC₅₀ values in the range between 5 and 22 μg/mL. The most active oil was that from Copaifera reticulata (collected in Pará State, Brazil) with IC₅₀ values of 5, 15, and 20 μg/mL for promastigote, axenic amastigote and intracellular amastigote forms, respectively. Amphotericin B showed IC₅₀ of 0.058 and 0.231 μg/mL against promastigote and amastigote forms, respectively. Cytotoxicity assay showed that this copaiba oil obtained from Copaifera reticulata showed low cytotoxicity against J774G8 macrophages.

Conclusion

Copaiba oils showed significant activity against the parasite Leishmania amazonensis.     

Byrsonima intermedia A. Juss.: gastric and duodenal anti-ulcer, antimicrobial and antidiarrheal effects in experimental rodent models

Ethnopharmacological relevance

An ethnopharmacological survey indicated that the leaves of Byrsonima intermedia A. Juss. (Malpighiaceae), a medicinal species commonly found in the Brazilian Cerrado, can be used against gastroduodenal disorders, such as gastric ulcers and diarrhea.

Aim of the study

The objective of this study was to evaluate the effects of a methanolic extract of Byrsonima intermedia (MBI) leaves on gastric and duodenal ulcers and to assess the antimicrobial and antidiarrheal effects of this extract.

Material and methods

The anti-ulcerogenic effect of MBI was investigated with different ulcerogenic agents in rodents (mice and rats), including non-steroidal anti-inflammatory drug (NSAID), HCl/ethanol, pyloric ligature, absolute ethanol, cysteamine and ischemia–reperfusion. The gastroprotective effect of MBI was assessed by analysing the volume of gastric juice, pH, total acidity, mucus, NO, sulfhydryl compound, vanilloid receptor, glutathione (GSH) levels, and myeloperoxidase (MPO) activity in the gastric and duodenal mucosa. The gastric and duodenal healing effects of MBI were also evaluated during 7 or 14 days of treatment. The antidiarrheal action (measured by intestinal motility and diarrhea induced by castor oil) and anti-bacterial action of MBI against Staphylococcus aureus, Escherichia coli and Helicobacter pylori were also evaluated by microdilution methods.

Results

The phytochemical profile from MBI indicated the presence of phenolic acids, flavan-3-ols, oligomeric proanthocyanidins, and flavonoids. MBI (500 mg/kg, p.o.) significantly inhibited totally gastric and duodenal lesions (69%) and healed gastric (49% on 14 days) and duodenal lesions (45% on 7 and 14 days). The MBI exert gastroprotective action by participation of endogenous sulfhydryl compounds, vanilloid receptors and increase in GSH level to effective gastric and duodenal protection. MBI also displayed curative (42%) and preventive (49%) antidiarrheal effects by involvement of opiate receptors and also antimicrobial effects in vitro.

Conclusions

Byrsonima intermedia leaves present gastroprotective, healing and antidiarrheal activities, supporting previous claims that its traditional use can treat gastrointestinal disorders.     

Aqueous extracts and polysaccharides from Liquorice roots (Glycyrrhiza glabra L.) inhibit adhesion of Helicobacter pylori to human gastric mucosa

Aims

Aqueous extracts from the roots of Glycyrrhiza glabra L. (Fabaceae) are widely used for treatment of stomach ulcer. The clinical proven effects are related to the presence of anti-inflammatory 12-keto-triterpensaponins in the extracts. Apart from that the influence of Glycyrrhiza glabra extract on the bacterial adhesion of Helicobacter pylori to stomach tissue was to be investigated. Additionally the influence of Glycyrrhiza glabra secondary compounds on the bacterial adhesion of Porphyromonas gingivalis, a major pathogen for induction of periodontal inflammations was to be investigated.

Methodology

In vitro cytotoxicity against Helicobacter pylori was investigated by agar diffusion assay; antiadhesive properties of aqueous extract, raw polysaccharides and purified polysaccharide fractions was investigated by means of an in situ adhesion assay with FITC-labelled bacteria on tissue slides of human stomach resectates.

Results

Aqueous extract (1 mg/mL) of Glycyrrhiza glabra significantly inhibited the adhesion of Helicobacter pylori to human stomach tissue. This effect was related to the polysaccharides isolated from the extract, with one purified acidic fraction (0.25 SPB) as main active polymer. Purified polysaccharides did not exhibit direct cytotoxic effects against Helicobacter pylori and did not influence hemagglutination. Additionally raw polysaccharides from Glycyrrhiza glabra were shown to have strong antiadhesive effects against Porphyromonas gingivalis.

Conclusion

Aqueous extracts and polysaccharides from the roots of Glycyrrhiza glabra are strong antiadhesive systems, which may be used as potent tools for a further development of cytoprotective preparations with anti-infectious potential.     

Antiulcer effects of Achyrocline satureoides (Lam.) DC (Asteraceae) (Marcela), a folk medicine plant,in different experimental models

Ethnopharmacological relevance

Achyrocline satureoides is a medium-sized South American indigenous herb, commonly known as “Marcela” or “Macela”. The infusion obtained from inflorescences of this plant is widely used in Brazilian folk medicine as an antispasmodic, anti-inflammatory, hypoglycemic, and hypocholesterolemic, mainly to treat gastrointestinal disorders such as gastric ulcers and dyspepsia. However, the antiulcer properties of this species have not yet been fully studied.

Aim of the study

This study was conducted to evaluate and contribute to validating the antiulcer activity of hydroalcoholic extract of inflorescences of Achyrocline satureoides.

Materials and methods

The antiulcer assays were performed using the ethanol-induced ulcer, and nonsteroidal anti-inflammatory drug (NSAID)-induced ulcer protocols. Gastric secretion parameters were also evaluated (volume, pH and total acidity) by the pylorus ligated model, and the mucus in the gastric content was determined.

Results

In the ethanol-induced ulcer model, it was observed that the treatment with Achyrocline satureoides extract significantly reduced the lesion index by 75.1 ± 8.6, 85.0 ± 9.2, 86.6 ± 7.4 and 75.5 ± 5.3 for the groups treated with 100, 250 and 500 mg/kg of extract of inflorescences of Achyrocline satureoides and the positive control (omeprazole 30 mg/kg), respectively. Significant inhibition was also observed in the lesion index in the indomethacin-induced ulcer model, with decreases of 62.5 ± 7.1, 62.5 ± 6.1, 63.6 ± 5.5 and 96.2 ± 3.6 for the groups treated with 100, 250 and 500 mg/kg of extract and the positive control (cimetidine 100 mg/kg), respectively. The parameters of gastric secretion (pH, volume, [H⁺]) showed no alteration in the different doses of the treatment. On the other hand the treatment with the hydroalcoholic extract of Achyrocline satureoides (100, 250 and 500 mg/kg), significantly increased mucus production (p < 0.01), when compared with the groups treated with indomethacin (100 mg/kg), cimetidine (100 mg/kg) and negative control (vehicle). No signs of toxicity was observed in the acute toxicity study.

Conclusions

The results of the present study show that hydroalcoholic extract of Achyrocline satureoides displays antiulcer activity, as demonstrated by the significant inhibition of the formation of ulcers induced using different models. However, this activity appears not be related to the antisecretor mechanisms. Moreover, this work suggests that preparations obtained from Achyrocline satureoides could be used for the development of new phytotherapic drugs for the treatment of gastric ulcer.