Antimutagenic, antigenotoxic and antioxidant activities of phenolic-enriched extracts from Teucrium ramosissimum: combination with their phytochemical composition

The evaluation of the mutagenic and antimutagenic actions of extracts obtained from aerial part of Teucrium ramosissimum was assayed using the Salmonella typhimurium assay system. The effect of the same extracts on genotoxicity and SOS response induced by aflatoxin B(1) as well as nitrofurantoin was investigated in a bacterial assay system, i.e., the SOS chromotest with Escherichia coli PQ37. The different extracts showed no mutagenicity when tested with Salmonella typhimurium strains TA100, TA98 and TA1535 either with or without S9 mix. In contrast, our results prove that T. ramosissimum extracts possess antimutagenic effects against sodium azide, aflatoxin B1, benzo[a]pyrene and 4-nitro-o-phenylenediamine. Moreover, the T. ramosissimum tested extracts exhibited no genotoxicity either with or without the external S9 activation mixtures. However, all the extracts significantly decreased the genotoxicity induced by aflatoxin B(1) and nitrofurantoin. The result obtained by the Ames test confirms those of SOS chromotest. Antioxidant capacity of the tested extracts was evaluated using the enzymatic (xanthine/xanthine oxidase assay) and the non enzymatic (NBT/riboflavine, DPPH and ABTS assays) systems. All extracts exhibited high antioxidant activity except the chloroform and the methanol extracts in DPPH and NBT/riboflavine assays respectively. Our results underline the potential of T. ramosissimum to avoid mutations and also its antioxidant potential.     

Evaluation of the antimutagenic,antigenotoxic, and antioxidant activities of Eriobotrya japonica leaves

Abstract

Context: The leaves of Eriobotrya japonica (Thunb.) Lindl. (Rosaceae) are used in traditional medicine to treat inflammatory diseases. However, information about the antigenotoxic and antioxidant properties of its leaves remains to be elucidated.

Objective: The objective of this work was to evaluate the mutagenic/antimutagenic, genotoxic/antigenotoxic, and antioxidant potentials of aqueous and total oligomers flavonoid (TOF) extracts from E. japonica.

Materials and methods: The mutagenic/antimutagenic and genotoxic/antigenotoxic potentials of extracts (50, 250, and 500?µg/plate) were evaluated, respectively, by the Ames test with 48?h incubation and the SOS chromotest test with 2?h incubation. The antioxidant capacity of these extracts (ranging from 50 to 700?µg/mL) was tested using xanthine/xanthine oxidase and the deoxyribose assays.

Results: Eriobotrya japonica extracts showed neither mutagenic nor genotoxic effect. The highest protective effect against methyl methanesulfonate and 2-aminoanthracene was obtained in the presence of aqueous extract, with IC₅₀ values of 80 and 140?µg/plate, respectively, against S. typhimurium TA104. Moreover, this extract (500?µg/plate) was also able to reduce significantly the genotoxicity induced by nitrofurantoin and aflatoxin B1 with IC₅₀ values of 140 and 240?µg/assay, respectively. Likewise, aqueous and TOF extracts inhibited xanthine oxidase and superoxide anion formation with IC₅₀ values ranging from 45 to 95 and from 70 to 90?µg/mL, respectively. However, TOF extract is more efficient in inhibiting hydroxyl radical and chelating iron ion with IC₅₀ values of 140 and 400?µg/mL, respectively, when compared with the aqueous extract.

Conclusion: Eriobotrya japonica prevents the genotoxicity of some carcinogenic substances probably thanks to its antioxidant capacities.     

Leaf extracts from Teucrium ramosissimum protect against DNA damage in human lymphoblast cell K562 and enhance antioxidant,antigenotoxic and antiproliferative activity

The in vitro antioxidant, antigenotoxic and antiproliferative activities of Teucrium ramosissimum extracts were investigated. The antioxidant activities of the tested extracts were evaluated through three chemical assays: The Cupric reducing antioxidant capacity, the reducing power and the ferric reducing antioxidant power. TR1 fraction from methanol extract showed the best antioxidant activity evaluated by the CUPRAC, RP and FRAP assays with TEAC values of 4.04, 1.77 and 1.48 μM respectively compared to control. Yet, TR2 fraction exhibited the lowest antioxidant effect with a TEAC values of 1.97, 0.408 and 0.35 μM respectively. All the tested extracts were also found to be effective in protecting plasmid DNA against the strand breakage induced by hydroxyl radicals. Furthermore, the effects of T. ramosissimum extracts on cell proliferation were also examined. The cytotoxic study revealed that methanol extract significantly inhibited the proliferation of K562 cells (IC50 = 150 μg/mL). The antigenotoxic properties of these extracts were investigated by assessing the induction and inhibition of the genotoxicity induced by the direct-acting mutagen, hydrogen peroxide (H₂O₂), using an eukaryotic system; the “Comet assay.” The results showed that all the extracts inhibited the genotoxicity induced by H₂O₂, and particularly TR2 fraction (96.99%) and methanol extract (96.64%).The present study has demonstrated that T. ramosissimum extract possess potent antioxidant, antiproliferative and antigenotoxic activities, which could be derived from compounds such as flavonoids and polyphenols.     

Investigation of biological activity of polar extracts isolated from Phlomis crinita Cav ssp. mauritanica Munby

The lyophilized infusion, the methanol, the ethyl acetate, and the total oligomer flavonoid (TOF)-enriched extracts prepared from the dried leaves of Phlomis crinita Cav. ssp. mauritanica Munby were investigated for the contents of flavonoids, tannins, coumarines and steroids. Antibacterial activity was investigated toward five bacterial strains. An inhibitory effect was observed against Staphyllococcus aureus and Enterococcus feacalis, and the minimal inhibitory concentrations ranged from 2.5 to 5 mg/mL of extract. The tested extracts exhibit an important free radical scavenging activity toward the 1,1-diphenyl 2-picrylhydrazyl (DPPH) free radical; with IC₅₀ values of 30.5, 6, 32, and 31.5 μg/mL, respectively, in the presence of lyophilized infusion, the TOF, the methanol, and the ethyl acetate extracts. Genotoxic and antigenotoxic properties of the different extracts were studied by using the SOS chromotest with Escherichia coli PQ37. The lyophilized infusion and TOF extracts obtained from P. crinita ssp. mauritanica showed no genotoxicity, whereas methanol and ethyl acetate extracts are considered as marginally genotoxic. On the other hand, we showed that each extract inhibited the mutagenicity induced by aflatoxin B1 (AFB1) (10 μg/assay) and nifuroxazide (NF) (10 μg/assay). The ethyl acetate extract showed the strongest level of protection toward the genotoxicity induced by both directly and indirectly genotoxic NF and AFB1. These tests proved that the lyophilized infusion possesses an antiradical activity likewise, it showed no genotoxic effect; that is why we choose this extract to assess its antiulcerogenic activity by using an ethanol-induced ulcerogenesis model in the rat. This test demonstrates that 300 mg/kg of a P. crinita ssp. mauritanica lyophilized infusion was more effective than the reference compound, cimetidine.     

Evaluation of antioxidant and antigenotoxic activity of two flavonoids from Rhamnus alaternus L. (Rhamnaceae): Kaempferol 3-O-β-isorhamninoside and rhamnocitrin 3-O-β-isorhamninoside

The antioxidant activity of kaempferol 3-O-β-isorhamninoside (K3O-ir) and rhamnocitrin 3-O-β-isorhamninoside (R3O-ir), isolated from the leaves of Rhamnus alaternus L., was determined by the ability of each compound to inhibit NBT photoreduction and to scavenge the free radical ABTS⁺. Genotoxic and antigenotoxic activities were assessed using the SOS chromotest.At a concentration of 150 μg/assay the two compounds showed the most potent inhibitory activity against superoxide anion by respectively 80.4% and 85.6%. K3O-ir was a very potent radical scavenger with an IC₅₀ value of 18.75 μg/ml. Moreover, these two compounds exhibit an inhibitory activity against genotoxicity induced by nitrofurantoine and aflatoxine B1 using the SOS chromotest bacterial assay system in the presence of Escherichia coli PQ37 strain.In this study, we have also evaluated correlation between antigenotoxic and antioxidant effects of K3O-ir and R3O-ir. The highest correlation was showed with R3O-ir (r = 0.999).     

Antigenotoxic and Free Radical Scavenging Activities of Extracts from Moricandia arvensis

This study evaluates genotoxic and antigenotoxic effects of extracts from leaves of Moricandia arvensis, which are used in traditional cooking and medicines. Extracts showed no genotoxicity when tested with the SOS Chromotest using E. coli PQ37 and PQ35 strains, except for the total oligomers flavonoids enriched extract. Petroleum ether and methanol extracts are the most active in reducing nitrofurantoin genotoxicity, whereas methanol and total oligomers flavonoids enriched extracts showed the most important inhibitory effect of H₂O₂ genotoxicity. In addition, these two extracts showed important free radical scavenging activity toward the DPPH^· radical, whereas the chloroform extract exhibited the highest value of TEAC against ABTS^+· radical.     

Antioxidant activity and total phenolic content of ethanolic extract of Caesalpinia bonducella seeds

The aim of this study was to assess the in vitro potential of ethanolic extract of Caesalpinia bonducella seeds as a natural antioxidant. The DPPH activity of the extract (20, 40, 50, 100 and 200 μg/ml) was increased in a dose dependent manner, which was found in the range of 38.93–74.77% as compared to ascorbic acid (64.26–82.58%). The IC₅₀ values of ethanolic extract and ascorbic acid in DPPH radical scavenging assay were obtained to be 74.73 and 26.68 μg/ml, respectively. The ethanolic extract was also found to scavenge the superoxide generated by EDTA/NBT system. Measurement of total phenolic content of the ethanolic extract of C. bonducella was achieved using Folin–Ciocalteau reagent containing 62.50 mg/g of phenolic content, which was found significantly higher when compared to reference standard gallic acid. The ethanolic extract also inhibited the hydroxyl radical, nitric oxide, superoxide anions with IC₅₀ values of 109.85, 102.65 and 89.84 μg/ml, respectively. However, the IC₅₀ values for the standard ascorbic acid were noted to be 70.79, 65.98 and 36.68 μg/ml respectively. The results obtained in this study clearly indicate that C. bonducella has a significant potential to use as a natural antioxidant agent.     

Antioxidant,free radical scavenging activities of Salvia brachyantha and its protective effect against oxidative cardiac cell injury

In this study, antioxidant and free radical scavenging activities of an endemic Salvia species (Salvia brachyantha (Bordz) probed. was assessed in vitro using 1,1-diphenyl-2-picryhydrazyl (DPPH) radical, β-carotene linoleic acid, superoxide anion radical, hydroxyl radical, and reducing power assays. Regarding our data, the plant extract exhibited antioxidant and radical scavenging activities at different magnitudes of potency. In addition, this study was undertaken to assess whether methanol extract of S. brachyantha could increase the endogenous antioxidant enzymes in cells, and where such increased cellular defences could provide protection against oxidative cell injury. Pre treatment of rat heart cell lines with 100 μg/ml of plant extract for 24 h significantly prevented cell damage and enhanced activity of antioxidant enzymes induced by a treatment with xanthine/xanthine oxidase. Increased reactive oxygen species and cell apoptosis induced by xanthine/xanthine oxidase was dose-dependently prevented when cells were pre treated for 24 h with plant extract. These results indicated that S. brachyantha could protect against cell injury via induction of the antioxidant enzyme defences. The extract of this plant might be valuable antioxidant natural sources and seemed to be applicable in both healthy medicine and food industry.     

Occurrence of aflatoxins in mahua (Madhuca indica Gmel.) seeds: Synergistic effect of plant extracts on inhibition of Aspergillus flavus growth and aflatoxin production

Occurrence of aflatoxin in Madhuca indica Gmel. seeds was determined by competitive ELISA. Eighty percent of mahua seed samples were found to be contaminated with aflatoxin. Total aflatoxin content ranged from 115.35 to 400.54 ppb whereas the concentration of AFB₁ was in the range of 86.43 to 382.45 ppb. Mahua oil was extracted by cold press expeller and analysed for contamination of aflatoxin in both the oil and cake samples. Total aflatoxin and aflatoxin B₁ were 220.66 and 201.57 ppb in oil as compared to that in cake samples where it was 87.55 and 74.35 ppb, respectively. Various individual and combined plant extracts were evaluated for their efficacy against growth of Aspergillus flavus and aflatoxin production in vitro. Combination of botanicals were found to be more effective in controlling fungal growth and aflatoxin production than individual extracts. Results of the present study suggests that synergistic effect of plant extracts can be used for control of fungal growth and aflatoxin production. These natural plant products may successfully replace synthetic chemicals and provide an alternative method to protect mahua as well as other agricultural commodities of nutritional significance from toxigenic fungi such as A. flavus and aflatoxin production.     

Antioxidant and antidermatophytic activities of essential oil and extracts of Metasequoia glyptostroboides Miki ex Hu

This study was undertaken to assess the antioxidant and antidermatophytic potential of the essential oil and extracts (hexane, chloroform, ethyl acetate and methanol) of Metasequoia glyptostroboides Miki ex Hu. Antioxidant activity was evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The free radical scavenging activities of the oil and ethyl acetate extract were found to be superior (IC₅₀ = 9.1 and 14.24 μg/ml, respectively) as compared to butylatedhydroxyanisole (BHA), (IC₅₀ = 18.27 μg/ml). Also the ethyl acetate extract revealed the highest phenolic contents (93.26 mg/g of dry wt) as compared to the other extracts. Further, oil (1250 μg/disc) and extracts (1750 μg/disc) revealed 35.33–67.66 and 18.0–53.3% antidermatophytic effect, respectively, along with their respective MIC values (62.5–500 and 250–4000 μg/ml) against Trichophyton rubrum KCTC 6345, T. rubrum KCTC 6375, T. rubrum KCTC 6352, T. mentagrophytes KCTC 6085, T. mentagrophytes KCTC 6077, T. mentagrophytes KCTC 6316, Microsporum canis KCTC 6591, M. canis KCTC 6348 and M. canis KCTC 6349. The oil also had a strong detrimental effect on spore germination as well as concentration and time-dependent kinetic inhibition of M. canis KCTC 6591.     

Essential oil composition and antioxidant activities of Curcuma aromatica Salisb.

The chemical composition of the hydro-distilled essential oil from leaves of Curcuma aromatica Salisb. was analysed by GC–MS. Twenty-three compounds representing 94.29% of the total oil were identified. The antioxidant activities of the oil and various extracts of C. aromatica were evaluated by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radical-scavenging assays. The oil and methanol extract showed potent DPPH radical-scavenging activities (IC₅₀ = 14.45 and 16.58 μg/ml, respectively), which were higher than butylated hydroxyanisole (IC₅₀ = 18.27 μg/ml). The extracts also exhibited remarkable superoxide radical-scavenging activities (IC₅₀ = 22.6–45.27 μg/ml) and the activity in the methanol extract was superior to all other extracts (IC₅₀ = 22.6 μg/ml). Furthermore, the amount of total phenolic compounds was determined and its content in ethyl acetate extract was the highest as compared to other extracts. The results indicate that the oil and extracts of C. aromatica could serve as an important bio-resource of antioxidants for using in the food industries.     

Genotoxic and antimutagenic activities of extracts from pseudocereals in the Salmonella mutagenicity assay

Extracts of amaranth (Amaranthus L.), sorghum (Sorghumbicolor L.) and Japanese millet (Echinochloafrumentacea L.) were evaluated for mutagenicity in Salmonellatyphimurium strains TA98, TA100 and TA102. All three pseudocereal extracts were also assessed for their antimutagenic properties against the direct mutagens 2-nitrofluorene (2NF) for strain TA98, 3-(5-nitro-2-furyl)acrylic acid (5NFAA) for TA100 and H₂O₂ for TA102 strain and against the indirect mutagen aflatoxin B₁ (AFB₁). No mutagenicity was induced by any of the pseudocereal extracts when tested at concentrations as high as 50 mg/ml. All three extracts showed similar antimutagenicity against 5NFAA and no antimutagenicity against 2NF. The number of revertants induced by H₂O₂ extract was inhibited in order amaranth > Japanese milet > sorghum. All extracts were effective in the inhibition of mutagenic activity of aflatoxin B₁. The total polyphenol content as well as the amount of the flavonoids and phenolic acids as main component of polyphenolics were also determined.     

Scavenging capacity of strawberry tree (Arbutus unedo L.) leaves on free radicals

Despite strawberry tree (Arbutus unedo L.) leaves had a long use in traditional medicine due to its antiseptic, diuretic, astringent and depurative properties, the potential of their antioxidant activity are still lacking. Our study goals to assess the antioxidant and free radical scavenging potential of water, ethanol, methanol and diethyl ether extracts of A. unedo leaves. Total phenols content was achieved spectrophotometrically using Folin–Ciocalteau reagent with gallic acid as standard. Antioxidant activity was evaluated using three different methods: reducing power of iron (III)/ferricyanide complex assay, scavenging effect on DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals and scavenging effect on superoxide radicals by using the PMS–NADH–nitroblue tetrazolium system. Ethanol extracts of A. unedo leaves were the highest in reducing power (IC₅₀ 232.7 μg/mL) and DPPH scavenging effect (IC₅₀ 63.2 μg/mL) followed by water extracts (with IC₅₀ of 287.7 and 73.7 μg/mL, respectively); whereas diethyl ether extracts were the lowest. In the scavenging on superoxide radical assay, methanol extracts obtained the best results (IC₅₀ 6.9 μg/mL). For all the methods tested the antioxidant activity was concentration dependent. In accordance with antioxidant activity, highest total phenols content were found in ethanol, followed by water, methanol and diethyl ether extract. The results indicated that A. unedo leaves are a potential source of natural antioxidants.     

Assessment in vitro of the genotoxicity,antigenotoxicity and antioxidant of Ceratonia siliqua L. extracts in murine leukaemia cells L1210 by comet assay

Genotoxicity of Ceratonia siliqua extracts, was investigated by assessing their capacity to induce nucleus DNA degradation of murine leukaemia cells L1210, using the “Comet assay”. The ability of total oligomer flavonoids (TOF) and aqueous extracts to protect cell DNA against oxidative stress induced by H₂O₂, was performed by pre- co or post-treatment of cells with the before mentioned extracts for different periods preceding exposure to H₂O₂ stress. No significant genotoxic effect was detected at different exposure times, except at the lowest concentration of TOF extract (16.25 μg/ml). It appears that extracts decreased DNA damage, induced by H₂O₂.Both of TOF and aqueous extracts exhibited cellular antioxidant capacity, with EC₅₀ values of respectively <16.25 and < 35 μg/ml, as well as, a protective capacity against lipidperoxidation inducing using L1210 cells line as a cellular model. MDA inhibition percentages reached 88.43% and 90.52% with respectively 35.5 μg/ml of TOF extract and 70 μg/ml of aqueous extract.Antioxidant properties of carob leaf extracts revealed by our study make a good antioxidant protection and thus a good candidate as food addition component.     

Antioxidant activity and total phenolic content of methanol extracts of Ixora coccinea

Objective: To investigate the in vitro antioxidant activity of methanol extracts of Ixora coccinea L. (Rubiaceae) flower, leaf and stem.

Materials and methods: The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, total antioxidant capacity (TAC) and xanthine oxidase inhibition assay were carried out to evaluate the antioxidant potential of the extract. The IC₅₀ values were calculated for the DPPH and xanthine oxidase assays in order to evaluate the antioxidant efficiency of each of the I. coccinea extracts. The phenol contents were also determined.

Results: I. coccinea flowers revealed the best antioxidant property, presenting much lower IC₅₀ value (6.6?mg/mL for DPPH assay). The flower extract showed a significantly higher antioxidant capacity compared to the other extracts. Furthermore, the highest phenolic content (polyphenols) was found in the flower extract (210.55?±?6.31 µg GAE/mg extract). Moreover, I. coccinea extracts scavenged the superoxide radical generated by the xanthine/xanthine oxidase system. The xanthine oxidase inhibition activity was in the order of allopurinol > leaf > flower > stem with the percentage of inhibition ranged from 39.7% to 77.3% for the plant parts investigated. The highest phenolic contents (polyphenols) were found in the flower extracts (210.55?±?6.31 µg GAE/mg extract).

Conclusions: I. coccinea could be considered as a potential source of natural antioxidant.     

Screening of antimutagenicity via antioxidant activity in different extracts from the leaves of Acacia salicina from the center of Tunisia

The effect of extracts obtained from Acacia salicina on genotoxicity and SOS response induced by Benzo(a)pyrene (B[a]P) as well as nifuroxazide was investigated in a bacterial assay system, i.e., the SOS chromotest with Escherichia coli PQ37. Preparations obtained from the leaves of A. salicina exhibited no genotoxicity either with or without the external S9 activation mixture. However, all extracts significantly decreased the genotoxicity induced by (B[a]P) and nifuroxazide. Ethyl acetate, methanol and TOF extracts exhibited the highest inhibition level of the SOS response induced by the direct mutagen nifuroxazide. Whereas, aqueous, ethyl acetate and methanol extracts displayed the greatest level of protection towards the indirect mutagen, (B[a]P), induced response. In addition to their antigenotoxic activity, TOF, aqueous, methanol and chloroform extracts showed an important free radical scavenging activity towards the 1,1-diphenyl 2-picrylhydrazyl (DPPH) free radical. These extracts showed IC(50) value of 36, 73, 65, and 87μg/ml respectively. Taken together, our finding showed that A. salicina exhibits significant antioxidant and antigenotoxic activities.     

Chemical compositions,antifungal and antioxidant activities of essential oil and various extracts of Melodorum fruticosum L. flowers

This research presents the chemical composition antifungal and antioxidant activities of essential oils and various extracts from Melodorum fruticosum flowers. The essential oil composition of M. fruticosum flowers were investigated by GC–MS with 88 identified volatile constituents. Phenyl butanone, linalool, benzyl alcohol, α-cadinol, globulol and viridiflorol were found to be the major components, respectively. The dichloromethane extract played a major role as a remarkable fungicide according to their inhibition action against all tested pathogens followed by hexane extract, essential oil and methanol extract, respectively, along with their respective MIC values ranging from 125 to 1000 μg/ml. The dichloromethane extracts were also evaluated to be superior to all extracts tested with an IC₅₀ value of 87.6 μg/ml whereas other extracts showed their IC₅₀ values ranging from 100.13 to 194.50 μg/ml.     

A search for hepatoprotective activity of aqueous extract of Rhus coriaria L. against oxidative stress cytotoxicity

The protective effects of different concentrations of aqueous extract of Rhus coriaria L. fruit (75 and 100 μg/ml) and also gallic acid (100 μM) as one of its main components were examined against oxidative stress toxicity induced by cumene hydroperoxide (CHP) in isolated rat hepatocytes. Both extract concentrations and gallic acid (100 μM) significantly (P < 0.05) protected the hepatocyte against all oxidative stress markers including cell lysis, ROS generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Aqueous extracts of Rhus coriaria L. (75 and 100 μg/ml) were more effective than gallic acid (100 μM) in protecting hepatocytes against CHP induced lipid peroxidation (P < 0.05). On the other hand gallic acid (100 μM) acted more effective than aqueous extracts of Rhus coriaria L. (75 and 100 μg/ml) at preventing hepatocyte membrane lysis (P < 0.05). In addition H₂O₂ scavenging effect of both extract concentrations (75 and 100 μg/ml) were determined in hepatocytes and compared with gallic acid (100 μM). Gallic acid (100 μM) was more effective than aqueous extracts of Rhus coriaria L. (75 and 100 μg/ml) at H₂O₂ scavenging activity (P < 0.05).