EOB anti-mutagenic effect in vitro_972

By Roger Reed,2014-10-30 17:05
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EOB anti-mutagenic effect in vitro_972

EOB anti-mutagenic effect in vitro

     Abstract Objective To explore the role of bamboo leaf flavone health care. Methods auxotrophic Salmonella typhimurium reverse mutation test (Ames test) on the bamboo leaves flavonoids in vitro anti-mutagenic effects research.

    The results of EOB has significant anti-mutation effect, when

    the dose of 20 mg / petri dish when 2-AF, B [a] P and AFB1

    mutagenicity inhibition rate reached 70.5% or more, and showed a dose -- effect relationship. Conclusion bamboo leaves

    flavonoids anti-mutation effect primarily by blocking mutagens so that normal cell into a mutant cell achieved, but also including some to the mutation, but the promotion of the role of mutant cells to repair is not obvious.

     Key words bamboo leaves flavonoids Antimutagenic Ames test

     Abstract: ObjectiveTo discuss the health function of flavonoids from bamboo leaves (FBL). MethodsAntimutagenicities in vitro of FBL were studied by Ames test.ResultsFBL showed strong antimutagenic activities with inhibition rates of more than 70.5% to B [a] P ,2-AF and AFB1 at dose of

    20mg/plate.ConclusionThe desmutagenicities and bio-

    antimutagenicities of FBL coexisted, while the latter is the main way by FBL inhibits metagena.

     Key words: Flavonoids from bamboo leaves (FBL);

    Antimutagenicity; Ames test

     Dendrocalamus Dendrocalamus latiflorus also known as sweet bamboo, hemp August (Fujian), belong to Gramineae Dendrocalamus, is widely distributed in China's Fujian, Taiwan, Guangdong, Guangxi, Yunnan, Guizhou and Sichuan, south subtropical and tropical regions of the large-scale have

arisen Bamboo species [1]. Ma bamboo leaves large, wide-4 ~ 7

    cm or more, which is rich in nutrients, it is worthy of exploitation. Bamboo in China has a long history of food and

    medicine [2,3]. Bamboo leaf extract has anti-free radical,

    anti-oxidation, anti-aging, regulate blood lipids, block the nitrosation reaction, enhance the role of immunization and anti-bacterium [3]. Function of factors contained in bamboo

    leaves is mainly flavonoid glycosides and coumarin lactones, etc. [3].

     1 Materials and methods

     1.1 Materials

     Bamboo: taken from homes, Suining City, Sichuan Province, Ma Chuk Yuen District, Suining City, Sichuan Province, through the pines Botanical Garden Shusong Tao Senior Gardener identified as bamboo Dendrocalamus latifl-orus leaves.

     Test strains: Salmonella typhimurium histidine-deficient

    mutant strains TA97, TA98 (frameshift mutant strain) and TA100 (base substitution mutant strains): provided by the Chongqing Municipal CDC.

     Animal: Kunming mice: In order to BALB / C mice were pure, ?, ? in half, 8 to 10 weeks old, weighing 18 ~ 20g, Chongqing Institute of Chinese Medicine by the Experimental Animal Center.

     2 - amino-fluorene (2-AF), benzopyrene (B [a] P): Sigma

    Corporation Chu Pin.

     Aflatoxin B1 (AFB1): Chongqing Family Planning Scientific Institute.

     Polysulfone ultrafiltration membrane (molecular cut-off

    flow of 50 000 Da), the State Oceanic Administration, Hangzhou

    Water Treatment Device Center Chupin.

     Other conventional pharmaceutical chemical pure or AR.

     1.2 Methods

     1.2.1 Preparation of Ma bamboo flavonoids bamboo leaves 5

kg, crushed into meal, to 80% ethanol was extracted three

    times, 2 h / times. Extract combined ultrafiltration separation, collection of clear liquid vacuum concentrated by silica gel column chromatography CHCl3-MeOH gradient elution,

    and then by repeated silica gel or ODS column chromatography, obtained samples of bamboo leaf flavonoids.

     1.2.2 Ames test [4,5] using the standard plate incorporation Ames also known as Plate incorporation.

     Mutagenicity inhibition rate (%) = G-H/G-I × 100%

     Type of G-induced mutation in the positive control group

    back to change the number of colonies (with ? s, said, the

    same below);

     H-experimental group back to change the number of colonies;

     I-solvent control group, changed back to colonies.

     1.2.3 EOB mechanism of anti-mutation experiments [4 ~ 7]

    In this experiment, three kinds of ways to join materials subjects, adding amount of both 20 mg / petri dish, the test strain TA98.

     Mode 1, the objects and subjects mutagens also added to phosphate buffer (pH 7.4) in a water bath at 37 ? after 30

    min pre-culture by adding bacilli, the following steps according to "1.2.2" to.

     Mode 2, the subjects objects, mutagens and the bacilli also added to the living enzymes containing phosphate buffer (pH 7.4) in a water bath at 37 ? for pre-incubation 30 min,

    the following steps according to "1.2.2" to .

     Mode 3, the mutagens and the bacilli at the same time by adding phosphate buffer (pH 7.4) and live enzymes in a water bath at 37 ? for pre-incubation 30 min, then centrifuged, the supernatant discarded. Precipitate with phosphate buffer (pH 7.4) washing 2 times. The band will be washed bacilli to blend with the subjects, the following steps according to "1.2.2" to.

     1.2.4 Statistical Methods DPS statistical software and analysis of experimental data taking the average of three determinations. Reposted elsewhere in the paper for free download http://

     2 Results and discussion

     2.1 bamboo leaves flavonoids in vitro anti-mutagenic

    effect of dose - effect relationship between the results in

    Table 1. Table 1 bamboo leaves flavonoids in vitro anti-

    mutagenic effect of dose - effect relationship (omitted)

     Said the chances of regression equation y units (with the DPS software system will inhibit the rate is converted to probability units); x that the logarithm of dose

     From Table 1 we can see, in the experimental dose range, the three kinds of bamboo leaves flavonoids on the

    mutagenicity of mutagens inhibition dose - response

    relationship after logarithmic transformation, its logarithmic dose - response relationship was highly significant (P < 0.01) or significantly (P <0.05) linear positive correlation,

    indicating the dose - response relationship for the

    logarithmic curve relationship, that with the continuous increase in each petri dish in the same amount of material subjects the dose caused by anti - mutation increases

    gradually, but its effects are a gradual increase is much


     2.2 EOB pairs of three kinds of mutagens-induced

    inhibition of mutation can see from Table 2, in the TA97, TA98 and TA100, the three kinds of bamboo leaves flavonoids on the mutagenicity of mutagens were significantly inhibited, when

    the dose of of 20 mg / petri dish when its inhibitory rate reached 70.5% more than the mutagenicity of mutagens in TA97 strain of the order of inhibitory effect of AFB1

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