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Des sources du savoir aux médicaments du futur

 | 
Jacques Fleurentin
, 
Jean-Marie Pelt
, 
Guy Mazars

2. Études chimiques et pharmacologiques

Evaluation of the mutagenic and antimutagenic activities of saponins from Hedera colchica K. Koch

V Mshvildadze, M De Méo, G Dumenil, G Dekanosidze, E Kemertelidze, R Elias y G Balansard

Texto completo

  • 1 Institute of Pharmacochemistry, Academy of Science of Georgia 36, P.Sarajishvili St. 380059 Tbilis (...)

1Note portant sur l'auteur1

  • 2 Laboratoire of Biogenotoxicity and Mutagenese Environnementale, (EA 1784)

2Note portant sur l'auteur2

  • 3 Laboratoire of Biogenotoxicity and Mutagenese Environnementale, (EA 1784)

3Note portant sur l'auteur3

  • 4 Institute of Pharmacochemistry, Academy of Science of Georgia 36, P.Sarajishvili St. 380059 Tbilis (...)

4Note portant sur l'auteur4

  • 5 Institute of Pharmacochemistry, Academy of Science of Georgia 36, P.Sarajishvili St. 380059 Tbilis (...)

5Note portant sur l'auteur5

  • 6 Laboratoire of Pharmacognosie, Faculty of Pharmacy 27 Boulevard Jean Moulin 13385 Marseille cedex (...)

6Note portant sur l'auteur6

  • 7 Laboratoire of Pharmacognosie, Faculty of Pharmacy 27 Boulevard Jean Moulin 13385 Marseille cedex (...)

7Note portant sur l'auteur7

Introduction

8Hedera colchica K. Kock. (Araliaceae) is an endemic plant in Georgia. The leaves of this plant are used in traditional medicine as bronchospasmolitic, secretolytic and anti-inflammatory remedies. (Sakartvelos Floras, 1984; Rastitllnie and col., 1988)

9The mutagenic and antimutagenic activities of 7 monodesmoside glycosides as well as the less polar glycoside fraction, were evaluated by using a modified version Ames test.

Material and methods

Extraction and Isolation

10Leaves and berries of Hedera colchica were collected in the Bagdathi region of Georgia.

11Powdered plant materials were extracted with MeH-H20 (80:20). After concentration, the aqueous layer was treated with CHCI3 then, with BuOH to obtain a crude extract of saponins. Pure compounds were isolated from the BuOH extract by the combination of different chromatographic technics. Structure elucidation were established using spectroscopic methods FAB-MS and 1H, 13C NMR (Mshvildadze and col., in press).

12Salmonella mutagenicity test

13• Tester strains

14Salmonella typhimurium TA97a, TA98, TA100,TA102 and YG1041.

15• Activation mixture S9mix

16The liver homogenate (S9) was prepared from Sprague-Dawley rats treated with Araclor 1 254 (500 mg/kg body weight). The protein concentration was 30.7 mg/ml (Lowry and col., 1951). The S9mix was a mixture of 4 % S9 and a solution of cofactors (Maron and col., 1983).

17• Assay procedure

18All samples were assessed for mutagenicity and antimutagenicity by a modified version the liquid incubation assay of the Ames test (Maron and col., 1983; De Meo and col., 1996).

19• Determination of the mutagenic and antimutagenic activities

20The mutagenic activity (MA) and the antimutagenic activity (AA) were calculated by non linear regression analysis using the Statgraphics Plus Software (STSC, Uniware, version 6.1) (De Meo and col., 1996).

Results

21The mutagenic and the antimutagenic activities of tested saponins (Fig. 1) were evaluated by using the Ames test. Assessment of the mutagenic activities was performed using tester strains TA97a, TA98, TA100 and TA102 with and without the metabolic fraction S9mix. The antimutagenic activity was performed against two known mutagens: BaP (benzo(a)pyrene) and a complex mixture CSC (Cigarette Smoke Concentrate)

22—> No mutagenic activity could be detected.

23The antimutagenic activities, expressed as ID50 were evaluated against 2 known mutagenic compounds. Results are summarized in Tables I and II. Typical dose-response curves are included in Fig. 2. No antimutagenic activity could be detected against BaP. The sapo- nosids 3 and 5 showed a weak antimutagenic activity against CSC (Table II).

Figure 1. Tested saponins: Crude extract of saponins (n° 6) and 7 monodesmosides (n° 1 to 8) were tested for their mutagenic and antimutagenic activities.

Figure 1. Tested saponins: Crude extract of saponins (n° 6) and 7 monodesmosides (n° 1 to 8) were tested for their mutagenic and antimutagenic activities.

Glc: Glucose Ara: Arabinose Glu: ac. Glucuronique Rha: Rhamnose

Figure 2. Antimutagenic activities of Colchiside 7 (3) and Colchiside 9 (5) against Cigarette Smoke Concentrate (CSC).

Figure 2. Antimutagenic activities of Colchiside 7 (3) and Colchiside 9 (5) against Cigarette Smoke Concentrate (CSC).

The effect of 3 and 5 on the mutagenic activity of CSC (2.5 (µg) was determined in the Ames test with tester strain YG1041 + S9mix.
The mutagenic activities of CSC were 844 ± 14. Saponosid 3 showed a synergistic effect µp to 20 µg/plate.

Table I. Antimutagenicity of the 14 saponosids against BaP (0.5 µg) using TA100+S9mix

Table I. Antimutagenicity of the 14 saponosids against BaP (0.5 µg) using TA100+S9mix

Table II. Antimutagenicity of the 14 saponosids against cigarette smoke concentrate (2.5 μg) using YG1041 +S9mix

Table II. Antimutagenicity of the 14 saponosids against cigarette smoke concentrate (2.5 μg) using YG1041 +S9mix

Bibliografía

References

SAKARTVELOS FLORA (1984) Metsniereba, Tbilisi, IX, 124.

RASTITLLNIE (1988) Resursi SSSR, Nauka, Leningrad Fam. Rutaceae- Elaegnaceae, 4, 62.

MSHVILDADZE V.D., FAVEL A., DELMAS F., ELIAS R., FAURE R., DEKANO- SIDZE G.E., KEMERTELIDZE E.P., BALANSARD G. (in press) Antifungol and antiprotozoal activities of saponins from Hedera colchica, Die Pharmazie, Ref. NP9629.

LOWRY O.H., ROSEBROUGH N.J., FARR A.L., RANDALL R.J. (1951) Protein measu rements with the folin-phenol reagent, J. Biol. Chem., 193, 265-275.

MARON D.M., AMES B.N. (1983) Revised methods for the Salmonella mutagenicity test, Mutation Res., 113, 173-215.

DE MÉO M.P, LAGET M., Dl GIORGIO C., GUIRAUD H., BOTTA A., CAS- TAGNARO M., DUMENIL G. (1996) Optimisation of the Salmonella/mam- malian microsome assay for urine mutagenesis by experimental designs, Mutation Res., 340, 51-65.

Notas finales

1 Institute of Pharmacochemistry, Academy of Science of Georgia 36, P.Sarajishvili St. 380059 Tbilisi (Georgia)

2 Laboratoire of Biogenotoxicity and Mutagenese Environnementale, (EA 1784)

3 Laboratoire of Biogenotoxicity and Mutagenese Environnementale, (EA 1784)

4 Institute of Pharmacochemistry, Academy of Science of Georgia 36, P.Sarajishvili St. 380059 Tbilisi (Georgia)

5 Institute of Pharmacochemistry, Academy of Science of Georgia 36, P.Sarajishvili St. 380059 Tbilisi (Georgia)

6 Laboratoire of Pharmacognosie, Faculty of Pharmacy 27 Boulevard Jean Moulin 13385 Marseille cedex 5 (France).

7 Laboratoire of Pharmacognosie, Faculty of Pharmacy 27 Boulevard Jean Moulin 13385 Marseille cedex 5 (France).

Índice de ilustraciones

Título Figure 1. Tested saponins: Crude extract of saponins (n° 6) and 7 monodesmosides (n° 1 to 8) were tested for their mutagenic and antimutagenic activities.
Leyenda Glc: Glucose Ara: Arabinose Glu: ac. Glucuronique Rha: Rhamnose
URL http://books.openedition.org/irdeditions/docannexe/image/7263/img-1.jpg
Archivo image/jpeg, 196k
Título Figure 2. Antimutagenic activities of Colchiside 7 (3) and Colchiside 9 (5) against Cigarette Smoke Concentrate (CSC).
Leyenda The effect of 3 and 5 on the mutagenic activity of CSC (2.5 (µg) was determined in the Ames test with tester strain YG1041 + S9mix.The mutagenic activities of CSC were 844 ± 14. Saponosid 3 showed a synergistic effect µp to 20 µg/plate.
URL http://books.openedition.org/irdeditions/docannexe/image/7263/img-2.jpg
Archivo image/jpeg, 111k
Título Table I. Antimutagenicity of the 14 saponosids against BaP (0.5 µg) using TA100+S9mix
URL http://books.openedition.org/irdeditions/docannexe/image/7263/img-3.jpg
Archivo image/jpeg, 444k
Título Table II. Antimutagenicity of the 14 saponosids against cigarette smoke concentrate (2.5 μg) using YG1041 +S9mix
URL http://books.openedition.org/irdeditions/docannexe/image/7263/img-4.jpg
Archivo image/jpeg, 435k

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