Antimutagenic properties of benzodiazapine tranquilizers

Translated from Khimiko-farmatsevticheskii Zhurnal, Vol. 23, No. 7, pp. 784–786, July, 1989.     

Antimutagenic effect of black tea extract using 'rodent dominant lethal mutation assay'.

The antimutagenic effect of black tea extract has been evaluated with the 'Dominant Lethal Assay' in Swiss albino mice using benzo[a]pyrene [BaP] as a mutagen. BaP was given through the intraperitoneal (i.p.) route at a single dose of 100 mg/kg b.w. to male mice once only. The animals were given 1, 2 and 4% aqueous solution of black tea as sole source of drinking solution prior to BaP. The pregnant females were analyzed for living implants, pre- and post-implantation losses. The results revealed that during mating weeks, BaP caused a reduction in implants and an increase in pre- and post-implantation losses. The protective effect of tea solution on BaP-induced mutagenicity was observed. The number of living implants increased and dead implants decreased significantly in the animals kept on 2 and 4% tea solution. The increase in dominant lethal mutation rate by BaP was inhibited by black tea extract. Four percent tea solution alone did not produce dominant lethality, and reveals that it is non-toxic/non-mutagenic to sperm. Hence the study suggests that tea has a protective effect against BaP-induced genetic damage to germ cells in Swiss albino mice.     

Antimutagenic properties of affinin isolated from Heliopsis longipes extract

Abstract

Context: Heliopsis longipes (A. Gray) Blake (Asteraceae), commonly known in Mexico as “chilcuage” or “chilcuan”, is widely used as an analgesic and anesthetic agent. Affinin, the major metabolite of this plant, and the ethanol extract of the plant have shown antinociceptive properties in mice. H. longipes plant produces a complex mixture of antioxidant chlorophylls and polyamines as well as a number of possible antimutagens.

Objective: The current study evaluated the potential utilization of the natural product affinin isolated from H. longipes ethanol extract as an antimutagenic and possibly anticarcinogenic agent.

Materials and methods: The Ames assay was used to assess the mutagenic properties of affinin (12.5, 25 and 50?µg/plate) that was added to several mutagens with or without S9 metabolic activation in Salmonella typhimurium (TA98, TA100 and TA102 strains).

Results: Heliopsis longipes extract and affinin were not toxic as a reduction in the number of His⁺ revertant bacteria colonies. Affinin (25 and 50?µg/plate) significantly reduced the frameshift mutations that were generated by 2-aminoanthracene (2AA) (40%) and reduced the oxidative DNA damage generated by norfloxacin (NOR) (37–50%). Affinin possessed antioxidant properties that were able to reduce 2AA- and NOR-induced mutations in S. typhimurium TA98 and TA102, respectively.

Discussion and conclusion: Affinin, the principal metabolite of H. longipes, is not mutagenic and possesses antimutagenic activity. These plants are currently used to treat some pain symptoms in Mexico; and antimutagen activity determined could be important to treat some pain symptoms related to antiradical activity.     

Antimutagenic activities of acetone and methanol fractions of Terminalia arjuna.

The antimutagenic effect of benzene, chloroform, acetone and methanol fractions from Terminalia arjuna, a well-known medicinal plant, was determined against Acid Black dye, 2-aminofluorene (2AF) and 4-nitro-o-phenylenediamine (NPD) in TA98 Frameshift mutagen tester strain of Salmonella typhimurium. Among the different fractions, the antimutagenic effect of acetone and methanol fractions was more than that observed with other fractions. Co-incubation and pre-incubation modes of experimentation did not show much difference in the antimutagenic activity of the extracts. Moreover, these fractions inhibited the S9-dependent mutagens, 2AF and Acid Black dye more effectively than the direct-acting mutagens. Studies are under way to isolate and elucidate the nature of the antimutagenic factor in acetone and methanol fractions.     

Antimutagenic effect of polysaccharide ginsan extracted from Panax ginseng.

Ginsan is a polysaccharide extracted from the roots of Panax ginseng, and it has earlier been reported to have an immunostimulatory effect. In the present study, the frequency of micronucleated polychromatic erythrocytes (MNPCE) was assessed in the bone marrow of C57BL/6 male mice treated with ginsan [100, 200 or 300 mg/kg body weight (b.w.)] or amifostine (200mg/kg b.w.) 30 min before as well as 15 min after 1.5 Gy of gamma-irradiation. Ginsan and amifostine did not alter the frequency of MNPCE of control mice (P>0.05), showing that they are non-mutagenic per se; gamma-irradiation induced a statistically significant (P<0.001) increase of MNPCE and decrease of PCE/NCE ratio (P<0.001) compared to control group. However, ginsan applied 30 min before or 15 min after irradiation reduced MNPCE in a dose-dependent manner. Amifostine (200mg/kg b.w.) did not reduce radiation-induced MNPCE, but stimulated erythropoiesis, when administered before irradiation. Based on the above results, radioprotective effect of ginsan can be partially attributed to reduction of radiation-induced genotoxicity.     

Antimutagenic effect of Maillard browning products obtained from amino acids and sugars.

The antimutagenic effects of Maillard reaction products (MRPs) prepared by heating three sugars (fructose, glucose and xylose) and four amino acids (arginine, glycine, lysine and tryptophan) at 100 degrees C for 10 hr was evaluated in the Salmonella/microsome assay. The highest extent of browning was found in the MRPs of sugars-lysine and xylose-amino acids. The MRPs of xylose-amino acids showed stronger antioxidative activity and reducing power than did the other combinations. No mutagenicity or toxicity in Salmonella typhimurium TA98 was observed with any of the MRPs in the presence of S-9. Most MRPs, especially those of sugars-tryptophan and xylose-amino acids, strongly inhibited the mutagenicity of 2-amino-3-methylimidazo(4,5-f)quinoline (IQ), 3-amino-1,4-dimethyl-5H-pyridol-(4,3-b)indole (Trp-P-1) and 2-amino-6-methyldipyrido(1,2-a:3',2'-d)imidazole (Glu-P-1) in the presence of S-9. However, the MRPs of fructose-glycine and fructose-arginine increased the mutagenicity of Trp-P-1. The antimutagenic effect of the MRPs was well correlated with their antioxidative activity and reducing power. The mutagenicity of benzo[a]pyrene was moderately inhibited by most MRPs, but was increased by the MRP of glucose-arginine. Aflatoxin B1 mutagenicity was increased greatly by all the MRPs except that of xylose-tryptophan. The findings suggested that MRPs might have a bifunctional property of co-mutagenicity and antimutagenicity in certain cases.     

Influence of drying and cooking process on the phytochemical content,antioxidant and hypoglycaemic properties of two bell Capsicum annum L. cultivars

The present study evaluates the influence of drying and cooking processes on the health properties of two bell Capsicum annuum L. cultivars Roggiano and Senise compared with fresh peppers. The content of phytochemicals decreased in the order fresh > dried > dried frying processes. HPLC analysis was applied to quantify five flavonoids from peppers. Apigenin was identified as main constituent. Its content was affected by drying and dried frying processes. The antioxidant activity was evaluated by DPPH, ABTS, β-carotene bleaching test and Fe-chelating activity assay. A comparable radical scavenging activity was observed for both cultivars. Interestingly, frying process did not influenced this property. Roggiano peppers exhibited the highest antioxidant activity using β-carotene bleaching test with IC₅₀ values of 38.1 and 24.9 μg/mL for total extract and n-hexane fraction, respectively. GC–MS analysis of lipophilic fraction revealed the presence of fatty acids and vitamin E as major components. In the inhibition of the carbohydrate-hydrolyzing enzymes fresh Senise peppers exerted the strongest activity against α-amylase with an IC₅₀ value of 55.3 μg/mL. Our results indicate that C. annuum cultivars Roggiano and Senise have an interestingly potential health benefits not influenced by processes that are used before consumption.     

Biochemical properties of polysaccharides from black pepper

The purified polysaccharides from Piper nigrum were prepared as follows: a hot water extract of pepper seeds was fractionated by ultrafiltration with a 5-kDa-membrane cartridge. A fraction with 5 kDa or bigger molecules was successively purified by open column chromatography on DEAE-Toyopearl 650C and Bio-gel P-60 with each active fraction, resulting in PN-Ib and PN-IIa, purified anti-complementary polysaccharides. None of the anti-complementary activity of any polysaccharide was changed by pronase digestion or polymyxin B treatment, but they were decreased by periodate oxidation. Analysis of component sugar and molecular mass determination of the anti-complementary polysaccharides indicated that PN-Ib with an average molecular mass of 21 kDa contained 88.5% glucose and other negligible minor monosaccharides, while PN-IIa showed a different monosaccharide composition, which contained a significant proportion of galactose, arabinose, galacturonic acid and rhamnose. The molar ratio of galactose and arabinose of PN-IIa (48 kDa) was 1.93:1. PN-1 did not react with beta-glucosyl Yariv reagent, however, PN-IIa did react, which indicated that PN-IIa might be an arabinogalactan. Based upon these results, the usefulness of purified anti-complementary polysaccharides from Piper nigrum is suggested as a supplement for immune enhancement.     

Composition analysis and antioxidant properties of black garlic extract

Black garlic produced from fresh garlic under controlled high temperature and humidity has strong antioxidant properties. To determine these compounds, five fractions (from F1 to F5) were separated and purified by elution with chloroform:methanol at different ratios (8:1, 6:1, 4:1, 2:1, and 0:1; v/v). The antioxidant activity of each fraction was analyzed. The results showed that F3 and F4 had higher phenolic contents and stronger 2,2-diphenyl-2-picrylhydrazyl radical scavenging activity than the others. Seven purified individual components were further separated using semipreparation high-performance liquid chromatography from these two intensely antioxidant fractions (F3 and F4), their structures were elucidated by high-performance liquid chromatography coupled to diode array detection, electrospray ionization, mass spectrometry, ¹H nuclear magnetic resonance, and ¹³C nuclear magnetic resonance spectrometry. Three compounds including adenosine, uridine, and 2-acetylpyrrole were first identified in black garlic, except for 5-hydroxymethylfurfural, (1S, 3S)-1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid, and (1R, 3S)-1-methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid. The cellular antioxidant activities of uridine, adenosine, carboline alkaloids, 5-hydroxymethylfurfural, and ethyl acetate extracts were consistent with the results of in vitro experimental antioxidant properties. The results provide useful information for understanding the health benefits of black garlic products.     

Purification and properties of hyaluronidase from Palamneus gravimanus (Indian black scorpion) venom.

Scorpion venoms are a rich source of enzymes. Some of the enzymes such as phospholipase A2, proteolytic enzymes and phosphodiesterase are well characterized. However, hyaluronidase has not been studied extensively. In this paper we describe the purification and characterization of hyaluronidase (Hyaluronate lyase, E.C.3.2.1.35) from the Palamneus gravimanus scorpion venom by a combination of gel filtration on Sephadex G-75 and ion-exchange chromatography on DEAE-cellulose. The optimal pH and temperature for its maximum activity of the isolated enzyme were 4.5 and 37 degrees C, respectively, and its K(m) was 47.61 microg/ml at 37 degrees C and its specific activity was 6411.7 +/- 117TRU/min per mg against 250 +/- 4.0 TRU/min per mg for the whole desiccated venom suggesting 25-fold purification. The molecular weight of the isolated enzyme was 52 +/- 1 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography on Sephadex G-75. The enzyme was stable for 30 days in the presence of NaCl; no loss of activity was observed up to 37 degrees degrees C and showed a sharp decrease in its activity at 40 degrees C. Heparin inhibited the enzyme activity.     

Antimutagenic and antioxidant activities of some bioflavours from wine

Monoterpenes limonene and its metabolic derivatives, α-terpineol and 1,8-cineol, commonly found as aroma wine components, were studied for their antimutagenicity by the bacterial reverse mutation assay on different strains. Substances were also tested for their antioxidant activity, i.e. radical scavenger, chelation, reduction, and lipid peroxidation inhibition. Limonene and its metabolites, α-terpineol and 1,8-cineol, resulted able to inhibit the chemically-induced mutagenesis, although with a different specificity. The antimutagenicity of limonene has been generally retained by its metabolites and sometimes increased. In particular, α-terpineol exhibited the strongest inhibition, moreover it showed to be a remarkable ferrous ions chelating agent. Limonene and 1,8-cineol were devoid of antioxidant activity. Present results are a starting point in evaluating the potential of α-terpineol as a chemopreventive agent and suggest potential functional dietary benefits of wine.     

Antioxidant properties of black tea in alcohol intoxication

Food ingredients such as alcohol may modify cellular redox state. Ethanol metabolism is accompanied by generation of free radicals that can damage cell components especially when antioxidant mechanisms are no able to neutralize them. However black tea is a source of polyphenol antioxidants that may enhance cellular antioxidant abilities. The aim of this study was to investigate the effect of black tea on antioxidant abilities of the liver, blood serum and brain of 12-months old rats sub-chronically (for 28 days) intoxicated with ethanol. Administration of black tea alone caused increase in the activity and concentration of antioxidant parameters more extensively in the liver and serum than in the brain. Alcohol caused decrease in the liver glutathione peroxidase and reductase and catalase activity but increase in activity of superoxide dismutase. Moreover, decrease in the level of non-enzymatic antioxidants, such as reduced glutathione, vitamin C, A and E and β-carotene was observed. The activity of serum glutathione peroxidase and reductase decreased while superoxide dismutase activity was not changed. The level of non-enzymatic antioxidants in serum was also decreased. However brain activity/level of all examined antioxidants enzymatic as well as non-enzymatic was decreased after ethanol intoxication. Black tea considerably prevented antioxidant parameters against changes caused by ethanol. These results indicate beneficial antioxidant effect of black tea regarding all examined tissues, but especially the liver.     

Antimutagenic and antioxidant characteristics of Chukrasia tabularis A Juss extracts

The current study aims to evaluate the antioxidative and antimutagenic activities of methanol extract and different fractions of Chukrasia tabularis leaves. The antioxidative potential was evaluated using 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cation and superoxide anion radical-scavenging assay. The antimutagenic potential was evaluated against direct-acting mutagens, 4-nitro-o-phenylenediamine and sodium azide; and S9-dependent mutagen, 2-aminofluorene in TA98 and TA100 strains of Salmonella typhimurium using Ames assay. It has been found that methanol extract and its fractions were more efficient against S9-dependent mutagen in pre-incubation mode of treatment as compared to direct-acting mutagens in both the strains. Methanol extract and its fractions also exhibited strong radical-scavenging potential. High-performance liquid chromatography (HPLC) analysis of methanol extract showed the presence of gallic acid, epicatechin, 7-hydroxycoumarin, and rutin. From the study, it could be concluded that antioxidative and antimutagenic activity of methanol extract and its fractions was related to the synergistic interactions among different chemical compounds.     

盐藻β—胡萝卜素抗突变效应

AIM: To study the genotoxic and antimutagenic efficts of beta-carotene from Dunaliella salina (beta-CDS). METHODS: The in vitro micronucleus and chromosomal aberration tests in hunman lymphocytes were adopted. The effect of beta-CDS on mutagensis induced by gamma-rays and mitomycin (Mit) was studied. RESULTS: beta-CDS (1-30 mg.L-1) had no genotoxicity, but inhibited spontaneous and gamma-ray-induced micronucleus fromation and Mit-induced chromosomal aberrations in human lymphocytes in vitro. The genotoxic action of synthetic beta-carotene (S beta C) was suppressed, the antimutagenic effects were heightened when S beta C and beta-carotene oil (beta CO, one of the beta-CDS compositions) were mixed in proportion as 80:27.5. CONCLUSION: beta-CDS is an antimutagenic agent.     

Antimutagenic and anti-oxidant activities of the non-steroidal anti-inflammatory drug celecoxib

1. Tumors arise and progress through the accumulation of serial genetic changes, including successive mutations, which involve activation of proto-oncogenes and inactivation of tumour suppressor genes, leading to the uncontrolled proliferation of progeny cells. The human body is continuously and unavoidably exposed to structurally diverse chemicals with established carcinogenic activity in animal models and/or mutagenic activity in short-term tests. 2. Celecoxib, a non-steroidal anti-inflammatory drug that specifically inhibits the enzyme cyclo-oxygenase-2, has been reported to be effective against certain types of cancers. The in vitro anti-oxidant and antimutagenic activities of the celecoxib were investigated in the present study using standard procedures. 3. The antimutagenic activity of celecoxib was determined using histidine mutant Salmonella typhimurium strains TA98, TA100, TA102 and TA1535 against directly acting mutagens (sodium azide (NaN3), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitro-o-phenylenediamine (NPDA) and doxorubicin) and mutagens needing activation (2-acetamidofluorene (2-AF) and 7,12-dimethylbenz [a] anthracene (DMBA)). 4. Celecoxib inhibited NaN3-, MNNG- and NPDA-induced mutations of TA100. The antimutagenicity of celecoxib (0.2 mg/plate) against the NaN3-induced mutation of TA1535 was 39.8% (P < 0.001). The MNNG-induced mutation of TA1535 was also inhibited by 0.3 mg/plate celecoxib (46.0%; P < 0.05). At concentrations of 0.2 mg/plate, celecoxib significantly inhibited NPDA- and doxorubicin-induced mutations of TA98 by 52.5 and 58.0%, respectively (P < 0.001 and P < 0.05, respectively). 5. The antimutagenic activity of 0.3 mg/plate celecoxib against 2-AF- and DMBA-induced mutations of TA98 was 81.76 and 98.1%, respectively (P < 0.001). 6. The anti-oxidant activity of celecoxib was determined by the inhibition of lipid peroxidation and superoxide and hydroxyl radical-scavenging activities. 7. The IC50 values of celecoxib for hydroxyl radical-scavenging and the inhibition of lipid peroxidation were 1.97 +/- 0.06 and 1.99 +/- 0.05 micromol/mL, respectively. Celecoxib had no superoxide radical scavenging-activity up to a concentration of 2.6 micromol/mL. 8. The in vitro antimutagenic and anti-oxidant activities of celecoxib indicate its possible therapeutic use as a cancer chemopreventive agent.     

Antimutagenic substances in the Armeniacae semen and Persicae semen]

Using the Ames/Salmonella/microsome assay, we examined the antimutagenic effect of the hexane extract of Armeniacae semen (apricot (Prunus armeniaca L.) seed), Persicae semen (peach (P. persica Bat.) seed), and seeds of cherry (P. avium L.), plum (P. salicina Lindle) and almond (P. dulcis Mill). Hexane extracts of Armeniacae semen and Persicae semen inhibited the mutagenicity of benzo[a]pyrene (B[a]P), but those of seeds of cherry, plum and almond did not. The mutagenicities of 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) and 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2) were also inhibited by the extracts of Armeniacae semen and Persicae semen. Inhibitory substances in Persicae semen were fractionated by silica gel column chromatography and high performance liquid chromatography, and were identified as oleic acid and linoleic acid. The contents of oleic acid and linoleic acid were 0.7 and 0.4% in the hexane extract of Armeniacae semen, and 1.5 and 0.5% in that of Persicae semen, respectively.     

Mutagenic and Antimutagenic Activities in Philippine Medicinal and Food Plants

Abstract

In developing countries there is increasing interest and research in the area of herbal medicines as an approach to reducing costs of health care. The chemically complex nature of these medicinal preparations results in a significant increased risk of toxicity, including genotoxicity. A total of 138 medicinal plant preparations used in the Philippines have been examined for genotoxicity using various short term bacterial and mammalian tests. Of the plants examined only the following 12 exhibited detectable genotoxicity in any system: Alli um sativum L., Aloe barbadensis Miller, Archangelisa flava (L.) Merr., Canarium luzonicum (Blume) A. Gray, Capsicum frutescens L., Entada phaseoliodes (L.) Merr., Moringa oleifera K., Nfrium indicum Mill., Piper betle L., Pithecellobium dulce (Roxb) Benth., Pittosporum pentandrum (Blanco) Merr., and Plantaqo major L. Little is known about the chemical nature of the mutagenic agents in these preparations.

Some plants also contain substances which reduce genotoxicity either by acting directly on the mutagen (desmutagens) or by acting on the affected organism (antimutagens). Examination of Philippine food and medicinal plants has identified numerous plants which contain antimutagenic activity. The chemical nature of the antimutagens has not been established but it has been speculated that part of the activity could be related to the vitamin content.     

Antimutagenic Activity of Essential Oil and Crude Extract of Phlomis fruticosa

The present study was carried out to investigate the antimutagenic activity of essential oil and crude extract of Phlomis fruticosa. This species belongs to the family Lamiaceae which includes a number of species with medical and pharmaceutical properties. The Escherichia coli K12 reversion assay for identifying antimutagens was used. The number of spontaneous and UV-induced Arg + revertants, as well as cell survival was monitored in the presence of non-toxic concentrations of the essential oil. The number of UV-induced revertants was slightly increased in the presence of essential oil. The ethanol extract demonstrated a significant effect on viable cells. Study on pharmacological and phytochemical activities as well as antimutagenetic potential against some chemical mutagens of the essential oils, total extract and their different fractions and constituents of Ph. fruticosa is in progress.