On the potential of Tephrosia purpurea as anti-Helicobacter pylori agent

Aim of the study

Since Tephrosia purpurea (Linn.) Pers. (Fabaceae) has traditional use in curing different types of wounds including gastroduodenal ulcers, it was of interest to evaluate the in vitro anti-Helicobacter pylori activity profile of the plant extract and its fractions with a view to examining its therapeutic potential, if any.

Materials and methods

Employing clinical isolates and standard strains of Helicobacter pylori, the extract and fractions were bioevaluated in terms of MIC and MBC values, acid stability, time-kill kinetics, drug resistance, and synergistic potential.

Results

The methanolic extract showed promising activity against clinical isolates and standard strains of Helicobacter pylori, including metronidazole-resistant strains. Fractionation of the extract revealed the n-hexane and chloroform fractions to possess marked activity. The extract and the less polar fractions remained functionally active in acidic condition similar to stomach environment, exhibited consistent bacteriostatic activity during repeated exposure, and demonstrated synergism, complete or partial, even with antibiotic-resistant strains.

Conclusion

Apolar fractions of Tephrosia purpurea may have therapeutic potential in combating Helicobacter pylori mediated gastroduodenal disorders.     

Evidence of the anti-Helicobacter pylori, gastroprotective and anti-inflammatory activities of Cuphea aequipetala infusion

Ethnopharmacological relevance

Cuphea aequipetala (Lythraceae) is a medicinal plant highly appreciated in Mexico to treat stomach ailments such as pain and burning sensation, stomach infections, ulcers, diarrhea, dysentery, and different types of tumors and bruises. In this work, the infusion of aerial parts of this plant (CAI) was investigated for its polypharmacological potential.

Materials and methods

In vitro anti-Helicobacter pylori activity was assessed by broth dilution method. Pharmacological studies included acute toxicity in mice using Lorke´s model, anti-inflammatory activity by xylene and TPA induced ear edema assay, as well as gastroprotection with ethanol-induced gastric ulcer model. DPPH and ABTS assays were used to determine antioxidant capacity. Polyphenols and flavonoid contents were determined by Folin–Ciocalteu method and AlCl₃ reaction, respectively.

Results

CAI showed good anti-Helicobacter pylori activity with a MIC of 125 μg/mL. The infusion was not toxic according to Lorke's model with a LD₅₀ greater than 5 g/kg. CAI exhibited low anti-edematogenic action in the models assayed. Oral administration of 300 mg/kg CAI significantly reduced gastric lesions by 87.9%. The effect was reversed only by indomethacin and N-ethylmaleimide demonstrating the role of endogenous prostaglandins and sulfhydryl compounds in gastroprotection. Total phenolic and flavonoid contents of CAI were 109.9 mg GAE/g DW and 28.1 mg QE/g DW, respectively, and the infusion exhibited a good antioxidant activity that is thought to play a role in its biological activity. The analysis of a preliminary fractionation of the infusion indicates that the complete extract conserves all its pharmacological activities in contrast to fractionated extracts.

Conclusions

Cuphea aequipetala is a promising native herb in an integral therapy for the treatment of bacterial or non-bacterial gastric ulcer because it possesses some anti-inflammatory properties, as well as exhibits good gastroprotective and antibacterial effects. It represents an important source for the isolation of anti-Helicobacter pylori compounds. This work provides ethnopharmacological evidence that supports the traditional use of this species.     

Anti-secretory,anti-inflammatory and anti-Helicobacter pylori activities of several fractions isolated from Piper carpunya Ruiz & Pav.

Ethnopharmacological relevance

The leaves of Piper carpunya Ruiz & Pav. (syn Piper lenticellosum C.D.C.) (Piperaceae), are widely used in folk medicine in tropical and subtropical countries of South America as an anti-inflammatory, anti-ulcer, anti-diarrheal and anti-parasitical remedy as well as an ailment for skin irritations.

Aims of the study

To study the anti-inflammatory, anti-secretory and anti-Helicobacter pylori activities of different fractions isolated from an ethanolic extract of the leaves of Piper carpunya, in order to provide evidence for the use of this plant as an anti-ulcer remedy. Moreover, to isolate the main compounds of the extract and relate their biological activity to the experimental results obtained with the fractions.

Materials and methods

Sixteen fractions were obtained from the ethanolic extract (F I–XVI) and 16 pure compounds were isolated and identified from these fractions. We studied the effects of the fractions (0.1–400 μg/mL) on the release of myeloperoxidase (MPO) enzyme from rat peritoneal leukocytes, on rabbit gastric microsomal H⁺, K⁺-ATPase activity and anti-Helicobacter pylori anti-microbial activity using the microdilution method (MM). The main compounds contained in the fractions were isolated and identified by ¹H- and ¹³C NMR spectra analysis and comparison with the literature data.

Results

Eight fractions showed inhibition of MPO enzyme (F I–IV, X, XII, XIV and XV). The highest inhibition was observed with F XIV (50 μg/mL, 60.9%, p < 0.001). F X and XII were the most active ones, inhibiting the gastric H⁺, K⁺-ATPase activity with IC₅₀ values equal to 22.3 μg/mL and 28.1 μg/mL, respectively. All fractions, except F XV, presented detectable anti-Helicobacter pylori activity, with a diameter of inhibition zones ranging from 11 mm up to 50 mm. The best anti-Helicobacter pylori activity was obtained with F III and V. Both fractions killed Helicobacter pylori with lowest concentration values, about 6.25 μg/mL. Sixteen pure compounds were isolated, five of them are flavonoids that possess strong anti-oxidant and free radical scavenging activity, e.g. vitexin, isovitexin, and rhamnopyranosylvitexin. Terpenoids like sitosterol, stigmasterol and phytol, which have shown gastroprotective activity, and dihydrochalcones, like asebogenin, with anti-bacterial activity, were also isolated. Furthermore, the rare neolignan 1, that is a DNA polymerase β lyase inhibitor, and (6S, 9S)-roseoside, that shows strong anti-bacterial activity, were isolated, for the first time, from the genus Piper.

Conclusions

We suggest that the flavonoids isolated from F I and II (vitexin, isovitexin, rhamnopyranosylvitexin and isoembigenin) contribute to the anti-MPO activity, as well as to their anti-Helicobacter pylori activity. These flavonoids may also be responsible for the important inhibition of H⁺, K⁺-ATPase activity. Also the phytosterols and phytol obtained from F XIV and XV could be involved in these gastroprotective activities. These results encourage us to continue phytochemical studies on these fractions in order to obtain full scientific validation for this species.     

In vitro and in vivo anti-Helicobacter pylori activity of Calophyllum brasiliense Camb.

Aims of the study

Calophyllum brasiliense (Camb.) is a medicinal tree that grows particularly in the hilly and forested regions of Brazil. Preparations from its stem bark are popular remedies for the treatment of chronic ulcers. Since earlier investigations on bark extracts evidenced gastroprotective and gastric acid inhibitory properties, this study evaluated the effects of hydroethanolic extract (HEECb) and the dichloromethanic fraction (DCMF), from Calophyllum brasiliense stem bark, against Helicobacter pylori, in vitro and in vivo.

Materials and methods

The in vitro assays were performed using the disk diffusion and broth microdilution methods to determine the minimum inhibitory concentration (MIC) values. The test substances were evaluated in vivo taking into account the delay in the gastric ulcer healing in Wistar rats, infected with Helicobacter pylori.

Results

DCMF appeared the most active and potent in vitro against Helicobacter pylori growth with an MIC of 31 μg/mL. In the in vivo assays, rats ulcerated by acetic acid, and inoculated with Helicobacter pylori showed a marked delay in ulcer healing. Treatment with HEECb (50, 100 and 200 mg/kg) and DCMF (100 and 200 mg/kg) reduced the ulcerated area in a dose-dependent manner. While DCMF, at 200 mg/kg, increased the prostaglandin E₂ (PGE₂) level, both HEECb and DCMF decreased the number of urease-positive animals, as confirmed by the reduction of Helicobacter pylori presence in histopathological analysis.

Conclusion

The results suggest that the antiulcer activity of Calophyllum brasiliense is due, in part, to its anti-Helicobacter pylori action, validating the popular use of this species.     

Qualea parviflora Mart.: An integrative study to validate the gastroprotective,antidiarrheal, antihemorragic and mutagenic action

Ethnopharmacological relevance

Qualea parviflora Mart. is a medicinal species commonly found in the Brazilian Cerrado biome.

Aim of the study

Based on ethnopharmacological data, methanolic extract from Qualea parviflora (QP) bark was evaluated for its antiulcer, analgesic, anti-hemorrhagic, mutagenic and anti-Helicobacter pylori activities.

Material and methods

The gastroprotective action of the extract was evaluated in rodent experimental models (HCl/ethanol, ethanol or NSAID). We also evaluated mutagenic effect (Ames assay), anti-Helicobacter pylori, anti-hemorrhagic action, analgesic and inflammatory effects (hot-plate test and carrageenin-induced hind paw edema) of methanolic extract from Qualea parviflora.

Results

QP (500 mg/kg, p.o.) was able to protect gastric mucosa against HCl/ethanol solution (77%), absolute ethanol (97%), and also against injurious effect of NSAID (36%). When QP was challenged with sulfhydryl depletor compound, the gastroprotective action of extract was abolished. QP treatment was able to maintain the GSH level and show a concentration-dependent inhibition effect on the lipid peroxidation. QP present anti-Helicobacter pylori effect (MIC = 75 μg/mL), anti-hemorrhagic and antidiarrheal action but not present analgesic or anti-inflammatory effect.

Conclusion

methanolic extract from Qualea parviflora had gastroprotective effect related to the increase of gastric mucosa defensive factors such PGE₂ levels and maintain the basal gastric glutathione levels. The methanolic extract also showed anti-Helicobacter pylori activity, anti-hemorrhagic effect and antioxidant action, but absence of analgesic, mutagenic and toxic effects, a profile that adds safety to its use.     

Mouririelliptica: Validation of gastroprotective,healing and anti-Helicobacter pylori effects

Ethnopharmacological relevance

Mouriri elliptica Martius (Melastomataceae) is species reputed in folk medicine to heal gastric ulcer and gastritis.

Aim of the study

Methanolic extract (ME) and ethyl acetate fraction (EAF) from leaves of Mouriri elliptica were evaluated for their gastroprotective, healing, immunological, toxicological and anti-Helicobacter pylori activities.

Material and methods

The gastroprotective action of ME and EAF was evaluated in rodent experimental models and to elucidate mechanisms of action, the antisecretory action, involvements of NO, SH, PGE₂, anti-Helicobacter pylori action of ME was evaluated. We also used immunohistochemical (PCNA and COX-2) and immunomodulatory (murine peritoneal macrophages) assays to evaluate Mouriri elliptica effects.

Results

ME present gastroprotective action without antisecretory effect. Otherwise, ME showed anti-Helicobacter pylori action (MIC = 0.025 μg/mL) and was able to inhibit NO production by macrophages. This species also accelerate the healing of ulcerated gastric mucosa by stimulating proliferation factors (PCNA), COX-2 and maintained basal PGE₂ level independent action of NSAID in gastric mucosa. The phytochemical investigation showed that this species possesses phenolic acid derivatives, acylglycoflavonoids and condensed tannins which probably influenced their pharmacological action.

Conclusion

All these results suggest the efficacy and safety of Mouriri elliptica in combating and healing gastric ulcer.     

Bactericidal activity of medicinal plants,employed for the treatment of gastrointestinal ailments,against Helicobacter pylori

Aim of the study

Helicobacter pylori infection plays a crucial role in the pathogenesis of peptic ulcer, and gastric cancer. The current PPI-based triple regimens for the eradication of Helicobacter pylori faces uprising resistance problem demanding for the search of novel candidates. Medicinal plants have always been a source of lead compounds for drug discovery.In the present study, we evaluated the anti-Helicobacter pylori activity of 50 commonly used Unani (traditional) medicine plants from Pakistan that are extensively utilized for the cure of gastrointestinal disorders to explore the natural source for pilot compounds against Helicobacter pylori.

Materials and methods

Total seven clinical isolates and one standard strain were employed to examine the bactericidal effects of medicinal plants. Helicobacter pylori was isolated from the antral biopsy specimens and confirmed through the standard microbiology procedures. Minimum bactericidal concentration (MBC) of the active plants was determined at the concentration range from 7.8 to 500 μg/ml.

Results

Among the herbs evaluated, more than 50% inhibited the growth of eight strains at the concentration of 500 μg/ml. The 70% aqueous-ethanol extracts of Curcuma amada Roxb., Mallotus phillipinesis (Lam) Muell., Myrisctica fragrans Houtt., and Psoralea corylifolia L. demonstrated strong anti-Helicobacter pylori activity with MBC value ranged from 15.6 to 62.5 μg/ml. The most potent bactericidal activity was exhibited by Mallotus phillipinesis (Lam) Muell. which completely killed the bacteria at the concentration of 15.6–31.2 μg/ml.

Conclusion

The results revealed significant anti-Helicobacter pylori activity of medicinal plants which could be the potential source of new bactericidal agents.     

Anti-Helicobacter pylori, gastroprotective,anti-inflammatory,and cytotoxic activities of methanolic extracts of five different populations of Hippocratea celastroides collected in Mexico

Ethnopharmacological relevance

The root barks of Hippocratea celastroides have been used for decades in Mexican traditional medicine to treat gastritis and ulcers. To investigate the anti-Helicobacter pylori, gastroprotective, anti-inflammatory, and cytotoxic activities of methanolic extracts obtained from the leaves, stems, and root bark of Hippocratea celastroides collected in five different localities in Mexico, during the winter of 2009, in order to establish differences in biological activities in terms of plant organs, as well as places of collection.

Materials and methods

Whole individuals were collected in five separate localities in Mexico: La Mancha, Veracruz (VL), Yautepec, Morelos (MY), Jojutla, Morelos (MJ), Temalac, Guerrero (GT), and Landa de Matamoros, Querétaro (QL). Methanolic crude extracts from wild plant specimens were tested using in vivo ethanol-induced mice gastric ulcer model, and 12-O-tetradecanoylphorbol-13-acetate (TPA) induced ear edema in mice assay, and in vitro anti-Helicobacter pylori model, and carcinoma cell line cytotoxic assays.

Results and conclusions

The leaves, stems, and root bark from MY specimens, as well as the leaves and root bark of materials from VL, presented the highest activity against Helicobacter pylori (MIC values ranging from 7.81 to 31.25 μg/ml). Most gastroprotective effects were displayed by the leaves of plants collected in MY, with 89.85±1.91% of protection (300 mg/kg) and an ED₅₀=27 mg/kg, which was corroborated by histological analysis. The root bark extracts from MY achieved the highest edema inhibition values (ED₅₀=0.18 mg/ear), which were comparable to indomethacin (ED₅₀=0.16 mg/ear). Finally, all extracts from MY (three plant parts) were cytotoxic against nasopharyngeal (KB), breast (MCF-7), and colon (HCT-116) carcinoma cell lines with IC₅₀ values between 1.18 and 9.77 μg/ml, except that no activity was detected for root bark extracts against HCT-116 normal fibroblasts. The activities of methanolic extracts from leaves, stems and root bark of plants collected in five different locations varied considerably, representing a notable problem facing the quality control of the plant material from Hippocratea celastroides used for medicinal purposes. The ethnomedical information of this plant in regards to treating gastritis and ulcers was strongly evidenced by the findings of the experimental models employed in this study.     

Anti-Helicobacter pylori activity of anacardic acids from Amphipterygium adstringens

Amphipterygium adstringens (Schltdl.) Standl. (Anacardiaceae) is widely used in traditional Mexican medicine for the treatment of gastritis and ulcers. In this work, we studied the anti-Helicobacter pylori activity of its bark, this Gram-negative bacterium is considered the major etiological agent of chronic active gastritis and peptic ulcer disease, and it is linked to gastric carcinoma. From a bio-guided assay of the fractions obtained form a continuous Soxhlet extraction of the bark, we identified that petroleum ether fraction had significant antimicrobial activity against Helicobacter pylori. From this fraction, we isolated an anacardic acids mixture and three known triterpenes: masticadienonic acid; 3α-hydroxymasticadienonic acid; 3-epi-oleanolic; as well as the sterol β-sitosterol. Only the anacardic acids mixture exhibits a potent dose-dependent antibacterial activity (MIC = 10 μg/ml in broth cultures). It is enriched in saturated alkyl phenolic acids (C_15:0, C_16:0, C_17:0 C_19:0) which represents a novel source of these compounds with potent anti-Helicobacter pylori activity. The promising use of anacardic acids and Amphipterygium adstringens bark in the development of an integral treatment of Helicobacter pylori diseases is discussed.     

Hepatoprotective and antioxidant effects of the methanolic extract from Halenia elliptica

Aims of study

Halenia elliptica, a medicinal herb of Tibetan origin, was commonly used in folk medicine to treat hepatitis. The aim of the present study is to evaluate the hepatoprotective and antioxidant activity of Halenia elliptica against experimentally induced liver injury.

Materials and methods

The antioxidant property of methanolic extract (ME) of Halenia elliptica was investigated by employing various established in vitro systems. The ME of Halenia elliptica was studied here for its hepatoprotective effects against CCl₄-induced liver toxicity in rats. Activity was measured by monitoring the levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and total bilirubin.

Results

The ME possessed strong antioxidant activity in vitro. The results of CCl₄-induced liver toxicity experiment showed that rats treated with the ME of Halenia elliptica (100 mg/kg and 200 mg/kg), and also the standard treatment, silymarin (50 mg/kg), showed a significant decrease in ALT, AST, ALP, and total bilirubin levels, which were all elevated in the CCl₄ group (p < 0.01). The results observed after administration of 100 mg/kg ME were comparable to those of silymarin at 50 mg/kg (p > 0.05). The ME did not show any mortality at doses up to 2000 g/kg body weight.

Conclusion

These results seem to support the traditional use of Halenia elliptica in pathologies involving hepatotoxicity, and the possible mechanism of this activity may be due to strong free radical-scavenging and antioxidant activities of ME.     

Bioevaluation of Anisomeles indica extracts and their inhibitory effects on Helicobacter pylori-mediated inflammation

Ethnopharmacological relevance

Helicobacter pylori is associated with the majority of gastric disorders and the antibiotic resistant rates have increased annually worldwide. Anisomeles indica and its constituent, ovatodiolide (OVT), were shown to have bactericide activity against Helicobacter pylori. The aim of this study was to manufacture extracts containing the effective constituent, OVT, and evaluate their bactericidal function and the inhibition of inflammatory responses to Helicobacter pylori infection.

Materials and methods

Various concentrations of ethanol for extraction of Anisomeles indica were performed and the content of OVT was analyzed by high-performance liquid chromatography (HPLC). The anti-bacterial activity of Anisomeles indica ethanol extracts and the constituent OVT were determined. Additional experiments were performed to investigate the Anisomeles indica ethanol extracts and OVT to inhibit the Helicobacter pylori-induced inflammation of both gastric epithelial cells and macrophages.

Results

Amongst the extracts tested, 50% and 95% ethanol extracts contained large amount of OVT and showed potent anti-Helicobacter pylori activity. An in vitro Helicobacter pylori-infection model revealed that 95% ethanol extract attenuated Helicobacter pylori-induced nuclear factor kappa B (NF-κB) activity and interleukin (IL)-8 secretion of gastric epithelial cells. In addition, 95% ethanol extract significantly inhibited lipopolysaccharide (LPS)-induced expression of inducible nitric oxide synthase (iNOS), as well as production of nitric oxide (NO) and tumor necrosis factor α (TNF-α) by macrophages.

Conclusions

This study reveals that Anisomeles indica ethanol extracts containing OVT may be a potent and economic therapeutic agent for Helicobacter pylori infection and attenuation of Helicobacter pylori-mediated inflammation.     

Anti-Helicobacter pylori activity of plants used in Mexican traditional medicine for gastrointestinal disorders

Aim of the study

Helicobacter pylori is the major etiological agent of chronic active gastritis and peptic ulcer disease and is linked to gastric carcinoma. Treatment to eradicate the bacteria failed in many cases, mainly due to antibiotic resistance, hence the necessity of developing better therapeutic regimens. Mexico has an enormous unexplored potential of medicinal plants. This work evaluates the in vitro anti-H. pylori activity of 53 plants used in Mexican traditional medicine for gastrointestinal disorders.

Materials and methods

To test the in vitro antibacterial activity, agar dilution and broth dilution methods were used for aqueous and methanolic extracts, respectively.

Results

Aqueous extracts of Artemisia ludoviciana subsp. mexicana, Cuphea aequipetala, Ludwigia repens,and Mentha × piperita (MIC 125 to <250 μg/ml) as well as methanolic extracts of Persea americana, Annona cherimola, Guaiacum coulteri, and Moussonia deppeana (MIC <7.5 to 15.6 μg/ml) showed the highest inhibitory effect.

Conclusions

The results contribute to understanding the mode of action of the studied medicinal plants and for detecting plants with high anti-Helicobacter pylori activity.     

Estimation of anti-inflammatory,antinociceptive and antioxidant activities on Arctium minus (Hill) Bernh. ssp. minus

Ethnopharmacological relevance

Arctium minus (Hill) Bernh. ssp. minus (Asteraceae) leaves are used to alleviate rheumatic pain, against fever and sunstroke with externally application in Turkish folk medicine.

Aim of the study

To evaluate the anti-inflammatory, antinociceptive and antioxidant activities of aqueous and ethanol extracts prepared from the leaves of Arctium minus ssp. minus.

Materials and methods

The ethanolic and aqueous extracts from the leaves of Arctium minus ssp. minus were evaluated in mice for anti-inflammatory activity using carrageenan-induced hind paw edema model and for antinociceptive activity using p-benzoquinone-induced abdominal contractions test. Moreover, the antioxidant power of the extracts has been determined by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and flow injection analysis-luminol chemiluminescence (FIA-CL). In addition, the total phenolic content in both extracts was determined with spectrophotometric method.

Results

Our results showed that only the ethanol extract exhibited a dose-dependent anti-inflammatory activity ranging between 11.1 and 23.6% at 200 mg/kg dose as well as displayed a significant antinociceptive activity without inducing any gastric damage. Although, both extracts were shown to possess significant DPPH radical-scavenging activity, that of aqueous extract was found to have more pronounced activity. In FIA-CL system, the ethanol extract was shown to possess a significant scavenger activity against H₂O₂ while the aqueous extract was much more potent antioxidant activity against HOCl-luminol CL than ethanol extract.

Conclusion

According to our results, it was concluded that Arctium minus ssp. minus contains potent natural antioxidants. In this study, in vivo experimental results have also supported the folk medicinal utilization of Arctium minus ssp. minus.     

Red ginseng extract inhibits the expression of MCP-1 and iNOS in Helicobacter pylori-infected gastric epithelial cells by suppressing the activation of NADPH oxidase and Jak2/Stat3

Ethnopharmcological relevance

Helicobacter pylori induced oxidative stress represents an important mechanism leading to expression of inflammatory mediators. Korean red ginseng is used in traditional medicine to inhibit inflammation. However, the anti-inflammatory mechanism of red ginseng is still under investigation. Thus, we investigated whether Korean red ginseng extract (RGE) inhibits NADPH oxidase, a source of reactive oxygen species (ROS), and the Jak2/Stat3 pathway, which mediates the expression of inflammatory mediators, in Helicobacter pylori-infected gastric epithelial cells.

Materials and methods

A standardized RGE was supplied by the Korea Ginseng Corporation. Human gastric epithelial cells (AGS) were treated with RGE and stimulated with Helicobacter pylori. NADPH oxidase activity, ROS levels, activation of Jak2/Stat3, and induction of MCP-1 and iNOS were determined.

Results

Helicobacter pylori infection resulted in an increase in ROS and activation of NADPH oxidase and Jak2/Stat3, which induced the expression of MCP-1 and iNOS in AGS cells. The induction of MCP-1 and iNOS was inhibited by both the Jak2/Stat3 inhibitor AG490 and RGE in Helicobacter pylori-infected cells. RGE suppressed NADPH oxidase activity by inhibiting translocation of cytosolic subunits p67phox and p47phox to the membrane and reduced ROS levels in Helicobacter pylori-infected cells.

Conclusion

RGE inhibits the expression of MCP-1 and iNOS by suppressing the activation of NADPH oxidase and Jak2/Stat3 in Helicobacter pylori-infected gastric epithelial cells.     

Chemical composition,antimicrobial activity against Staphylococcus aureus and a pro-apoptotic effect in SGC-7901 of the essential oil from Toona sinensis (A. Juss.) Roem. leaves

Ethnopharmacological relevance

Leaves of Toona sinensis (A. Juss.) Roem. (TSL), a popular vegetable in China, have anti-inflammatory, antidoting, and worm-killing effects and are used in folk medicine for the treatment of enteritis, dysentery, carbuncles, boils, and especially abdominal tumors. Our aim was to investigate the in vitro antimicrobial activity against Staphylococcus aureus and anticancer property of the essential oil from TSL (TSL-EO), especially the pro-apoptotic effect in SGC-7901.

Materials and methods

TSL-EO obtained by hydrodistillation was analyzed by GC/MS and was tested in vitro against twenty clinically isolated strains of Staphylococcus aureus (SA 1–20), which were either methicillin-sensitive Staphylococcus aureus (MSSA) or methicillin-resistant Staphylococcus aureus (MRSA) and two standard strains viz. ATCC 25923 and ATCC 43300. The anticancer activity of TSL-EO was evaluated in vitro against HepG2, SGC7901, and HT29 through MTT assay. Moreover, the apoptosis-inducing activity of TSL-EO in SGC7901 cells was determined by Hoechst 33324 staining and flow cytometry methods. Also, the apoptosis-related proteins viz. Bax, Bcl-2 and caspase-3 were detected by western-blotting.

Results

GC–MS analysis showed that TSL-EO contained a high amount of sesquiterpenes (84.64%), including copaene (8.27%), β-caryophyllene (10.16%), caryophyllene (13.18%) and β-eudesmene (5.06%). TSL-EO inhibited the growth of both MSSA and MRSA, with the lowest MIC values of 0.125 and 1 mg/ml, respectively. Treatment with TSL-EO for 24 h could significantly suppress the viability of three different cancer cell lines (P<0.05). Furthermore, the apoptosis-inducing activity of TSL-EO in SGC7901 cells increased in a dose-dependent manner, potentially resulting from the up-regulated expression of Bax, caspase-3 and down-regulated expression of Bcl-2.

Conclusions

TSL-EO possessed antibacterial activity against Staphylococcus aureus and significant cytotoxicity against cancer cells and particularly prominent pro-apoptotic activity in SGC7901 cells. These bioactivities were probably due to the high content of sesquiterpenes. Our results suggested that TSL-EO possessed potential health benefits and could serve as a promising natural food addictive.     

Antimicrobial,anti-inflammatory,anti-ulcer and antioxidant activities of Carlina acanthifolia root essential oil

The root of Carlina acanthifolia All. (Asteraceae) has been traditionally used in the treatment of various disorders including stomach and skin diseases. We studied antimicrobial, anti-inflammatory, anti-ulcer and antioxidant activities of Carlina acanthifolia root essential oil, in order to validate some of the ethnopharmacological claims. Antimicrobial activity was tested on 15 bacteria and three strains of fungi using the agar diffusion and broth microdilution methods. In assessing anti-inflammatory activity the carrageenan-induced rat paw oedema test was used, while ethanol-induced stress gastric ulcer test in rats was used in testing anti-ulcer activity. Antioxidant properties were evaluated trough the effect of the essential oil on lipid peroxidation (TBA assay) and its capability of quenching 2,2-diphenyl-1-picrylhydrazyl (DPPH) and OH radicals. The oil expressed significant antimicrobial activity, being the most active against Gram (+) bacteria: Streptococcus pyogenes, Enterococcus faecalis, Bacillus subtilis and against Candida albicans. In all applied concentrations, Carlina acanthifolia root essential oil reduced carrageenan-induced rat paw oedema in dose-dependent manner, achieving high degree of anti-inflammatory activity. The effect was comparable with that of indomethacin used as a reference drug. In the ethanol-induced stress gastric ulcer test in rats, it was shown that the tested essential oil produced significant dose-dependent gastroprotective activity. The results also pointed out substantial and dose-dependent antioxidant activity of the investigated essential oil, with carlina oxide as the main antioxidant component.     

Gastroprotective effect of Hintonia latiflora and Hintonia standleyana aqueous extracts and compounds

Ethnopharmacological relevance

The stem-barks of Hintonia latiflora and Hintonia standleyana, locally known as “copalchi”, are used for treating several maladies such as diabetes and gastrointestinal complaints, including gastric ulcers. Although the antidiabetic properties have been demonstrated, the gastroprotective action remains unexplored.

Aim of the study

The main goals of this study were to establish the potential acute toxicity and the gastroprotective activity of aqueous extracts and compounds from Hintonia latiflora and Hintonia standleyana in order to demonstrate their preclinical efficacy for the treatment of gastric ulcers in Mexican folk medicine.

Materials and methods

The aqueous extracts from the stem-barks (HLSB and HSSB) and leaves (HLL and HSL) from Hintonia latiflora and Hintonia standleyana were prepared by infusion. Investigation of the acute toxicity was accomplished by the Lorke method. The gastroprotective effect was assessed by means of a conventional ethanol-induced gastric injury model in rats using carbenoxolone as positive control. 5-O-[β-d-apiofuranosyl-(1→6)-β-d-glucopyranosyl]-7-methoxy-3′,4′-dihydroxy-4-phenylcoumarin (1) and chlorogenic acid (2) were also assayed. Preliminary mechanism of action of the tested compounds was analyzed using the same pharmacological models but pretreating the animals with N^G-nitro-l-arginine methyl ester (l-NAME), N-ethylmaleimide (NEM) and indomethacin.

Results

Investigation of the acute toxicity revealed that infusions of the leaves and stem-barks of both Hintonia species were not toxic to mice (LD₅₀>5000 mg/kg in all cases). HLSB, HSSB, HLL and HSL provoked a significant gastroprotective effect [80.5±3.35% (ED₅₀=184.7 mg/kg), 80.26±3.96%, 75.1±7.26% (ED₅₀=109.1 mg/kg), 76.85±3.17% (ED₅₀=149.7 mg/kg) of gastroprotection respectively]. Compounds 1 and 2, present in all the extracts, were also active [68.85±8.4% (ED₅₀=15 mg/kg), 74.04±4.4% (ED₅₀=26 mg/kg) of gastroprotection respectively] and their mode of action involved non-protein sulfhydryl endogenous (NP-SH) compounds, since only pretreatment with NEM inhibited their gastroprotective action.

Conclusions

The present investigation tends to support the ethnomedical use of HLSB, HSSB for treating gastric ulceration. Since HLL and HSL were also active, the leaves could be use alternatively, which in terms of natural resources conservation is an outstanding finding, considering that the plant populations of both Hintonia are scarce and in danger of extinction. Mainly two compounds (1 and 2) are important active principles of the plants.     

Screening of the topical anti-inflammatory activity of the bark of Acacia cornigera Willdenow, Byrsonima crassifolia Kunth, Sweetia panamensis Yakovlev and the leaves of Sphagneticola trilobata Hitchcock

Ethnopharmacological relevance

An investigation of topical anti-inflammatory activity was undertaken on plants used in Central America traditional medicine.

Aim of study

Four herbal drugs used in the folk medicine of Central America to treat inflammatory skin affections (Acacia cornigera bark, Byrsonima crassifolia bark, Sphagneticola trilobata leaves and Sweetia panamensis bark) were evaluated for their topical anti-inflammatory activity.

Materials and methods

Petroleum ether, chloroform and methanol extracts were obtained for herbal medicines and then extracts were tested on Croton oil-induced ear dermatitis model in mice.

Results

Almost all the extracts reduced the Croton oil-induced ear dermatitis in mice and the chloroform ones showed the highest activity, with ID₅₀ (dose giving 50% oedema inhibition) values ranging from 112 μg/cm² (Byrsonima crassifolia) to 183 μg/cm² (Sphagneticola trilobata). As reference, ID₅₀ of the non-steroidal anti-inflammatory drug indomethacin was 93 μg/cm².

Conclusions

Lipophilic extracts from these species can be regarded as potential sources of anti-inflammatory principles.     

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.     

Essential oils from Distichoselinum tenuifolium: Chemical composition,cytotoxicity, antifungal and anti-inflammatory properties

Ethnopharmacological relevance

Several aromatic plants and their essential oils are known to possess antimicrobial and anti-inflammatory properties. Distichoselinum tenuifolium (Lag.) Garcia Martin & Silvestre, an Iberian endemism, is traditionally used in the treatment of contact dermatitis and skin infections. However, the cellular mechanisms through which this plant exerts their beneficial effects are not known.

Aim of the study

The aims of this study were to examine the chemical composition of the essential oil isolated from Distichoselinum tenuifolium, and to test the efficacy of the essential oil as an antifungal and anti-inflammatory potential.

Materials and methods

The oils were investigated by gas chromatography (GC) and gas chromatography–mass spectrometry (GC–MS) and the antifungal activity (minimal inhibitory concentrations: MIC and minimal lethal concentrations: MLC) were evaluated against yeasts, dermatophyte and Aspergillus strains. Assessment of cell viability was made by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the in vitro anti-inflammatory potential of Distichoselinum tenuifolium oil was evaluated by measuring nitric oxide (NO) production induced by LPS, in the absence or in the presence of the oil, in a mouse macrophage cell line.

Results

The oils are predominantly composed of monoterpene hydrocarbons, being myrcene the main compound (47.7–84.6%). The oils revealed significant antifungal activity against Cryptococcus neoformans and dermatophyte strains and significantly inhibited NO production stimulated by LPS in macrophages, without affecting cell viability at concentrations ranging from 0.64 μL/mL to 1.25 μL/mL.

Conclusion

These findings add significant information to the pharmacological activity of Distichoselinum tenuifolium essential oils, specifically to its antifungal and anti-inflammatory properties, thus justifying and reinforcing the use of this plant on traditional medicine. Therefore, their beneficial effects and use in disease prevention, especially those related to fungal infections and inflammation, should be explored in more depth.