Mutat10n Re5earch, 241 (1990) 305-312

305

E15ev1er MU76EN 01565

M0d1f1cat10n 0f c1a5t09en1c1ty 0f 1ead and a1um1n1um 1n m0u5e 60ne marr0w ce115 6y d1etary 1n9e5t10n 0f Phy11anthu5em611cafru1t extract Ha1mant1 Dh1r, Aj0y Kumar R0y, Archana 5harma and 6eeta 7a1ukder Human 6enet1c5 Un1t, Centre 0f Ad0anced 5tudy 1n Ce11and Chr0m050meRe5earch, Department 0f 80tany, Un1ver51ty0f Ca1cutta, Ca1cutta-700 019 (1nd1a)

(Rece1ved16 Fe6ruary 1989) (Rev1510nrece1ved1 Fe6ruary 1990) (Accepted 7 Fe6ruary 1990)

Keyw0rd5:Phy11anthu5 em611ca:A5c0r61cac1d; C1a5t09en1e1ty;Lead; A1um1n1um

5ummary Extract 0f Phy11anthu5 em611ca fru1t and a5c0r61c ac1d were eva1uated 5eparate1y f0r pr0tect10n a9a1n5t c1a5t09en1c1ty 1nduced 6y 1ead (P6) and a1um1n1um (A1) 5a1t5 0n m0u5e 60ne marr0w chr0m050me5. 0ra1 adm1n15trat10n 0f Phy11anthu5 fru1t extract (PFE) f0r 7 day5 6ef0re exp05ure t0 60th meta15 6y 1ntraper1t0nea1 1nject10n 1ncrea5ed the fre4uency 0f ce11 d1v1510n and reduced the fre4uency 0f chr0m050me 6reak5 519n1f1cant1y. C0mpara61e d05e5 0f 5ynthet1c a5c0r61c ac1d (AA) were 1e55 effect1ve and c0u1d pr0tect a9a1n5t the effect5 0f A1 and 0n1y a 10w d05e 0f P6 (10 m 9 / k 9 60dy we19ht). AA adm1n15tered 6ef0re treatment 1n m1ce 91ven h19her d05e5 0f P6 (40 m 9 / k 9 60dy we19ht) enhanced the fre4uency 0f chr0m050me 6reak5, 91v1n9 a 5yner915t1c effect. 7he h19her pr0tect10n aff0rded 6y PFE may 6e due t0 the c0m61ned act10n 0f a11 1n9red1ent5, rather than t0 AA a10ne.

7he fru1t5 0f Phy11anthu5 em611ca L. ( = Em611ca 0ff1c1na115 6aertn.) (fam11y Euph0r61aceae) and 1t5 re1ated 5pec1e5 have 6een u5ed exten51ve1y 1n 1nd1an 1nd19en0u5 5y5tem5 0f med1c1ne f0r treat1n9 1euc0rrh0ea (Ra0 et a1., 1985), 5curvy (Ch0pra et a1., 1956) and u1cerat10n (6upta, 1908). 7he a4ue0u5 extract ha5 6een rec0rded t0 have ant1pyret1c, 1axat1ve and t0n1c pr0pert1e5, and a150 5h0wed ant16acter1a1 (V1naya9am00thy, 1982), ant1v1ra1 (5a190pa1 et a1., 1986) and ant10x1dat1ve act1v1t1e5 (Ra0 and 51dd14ue, 1964). 1n 9enera1, the5e act1v1C0rre5p0ndence: Dr. (Mr5.) H. Dh1r, Human 6enet1c5 un1t, Centre 0f Advanced5tudy 1n ce11ar1dChr0m050meRe5earch, Department 0f 80tany, un1ver51ty0f Ca1cutta, 35 8a11y9un9e C1rcu1arR0ad, Ca1cutta-700019 (1nd1a).

t1e5 have 6een attr16uted t0 the pre5ence 0f a5c0r61c ac1d (AA) and certa1n a1ka101d5 1n the extract. AA 1t5e1f, due t0 1t5 reduc1n9 and che1at1n9 act10n, exert5 a w1de var1ety 0f effect5 1nc1ud1n9 ant1muta9en1c and ant1c1a5t09en1c pr0pert1e5 (M1rv15h, 1975; Par5had et a1., 1978; 6e6hardt et a1., 1985; 61r1 and 8anerjee, 1986). 1t ha5 6een 5h0wn t0 anta90n15e the t0x1c act1v1ty 0f certa1n meta111c 5a1t5 1n mamma11an 5y5tem5 a5 we11 (Chakra6arty et a1., 1977). De5p1te the exten51ve med1c1na1 u5e 0f the fru1t5 0f th15 5pec1e5, very 11tt1e 15 kn0wn a60ut 1t5 r01e 1n pr0tect10n a9a1n5t effect5 0f t0x1cant5 0n chr0m050me5. 7he pre5ent w0rk wa5 undertaken w1th a 2-f01d 06ject1ve: (1) t0 a55e55 the pr0tect10n

0165-1218/90/$03.50 • 1990 E15ev1er5c1encePu6115her58.V. (810med1ca1D1v1510n)

306 aff0rded 6y extract 0f the fru1t5 0f Phy11anthu5 em611ca a9a1n5t the c1a5t09en1c act1v1ty 0f certa1n meta15; and (11) t0 c0mpare the pr0tect1ve r01e5 0f the extract and AA a10ne 1n c0mpara61e am0unt5 a9a1n5t the5e meta15• Mater1a15 and meth0d5

7he 0pt1mum d05e 0f AA 1n da11y d1et 15 c0ntr0ver51a1. Camer0n and Pau11n9 (1979) 5u99e5ted a da11y 1ntake 0f 1-10 9/day. Data 6a5ed 0n max1mum a5c0r6ate c0ncentrat10n5 1n the human 60dy 5u99e5t a max1mum 60dy p001 0f ar0und 5000 m9, 1.e., appr0x1mate1y 70 m 9 / k 9 60dy we19ht (6.w.) 1n man and 76 m 9 / k 9 6.w. 1n m0nkey5 (5ee C0un5e11 and H0rn19, 1981). 1n the pre5ent 5tudy, 16.66 m 9 / k 9 6.w. 0f AA wa5 u5ed f0r m1ce, a5 ca1cu1ated fr0m a da11y 1-9 1ntake f0r a 60-k9 per50n. A c0rre5p0nd1n9 am0unt 0f a4ue0u5 extract fr0m dr1ed fru1t5 0f Phy11anthu5 em611ca (685 m 9 / k 9 6.w.), c0nta1n1n9 the 5ame am0unt 0f AA, wa5 u5ed. 7he fru1t5 were pr0cured 1n 6u1k, cut 1nt0 5ma11 p1ece5, dr1ed 1n 5un119ht, kn0wn 4uant1t1e5 we19hed and kept 1n d15t111ed water f0r 24 h. 7he AA c0ntent 0f the dec0ct10n wa5 e5t1mated 6y the 2,6-d1ch10r0phen01 1nd0phen01 meth0d (Pear50n, 1952). E19ht-week-01d hea1thy, 1a60rat0ry-6red, 5w155 a161n0 m1ce (Mu5 mu5cu1u5), we19h1n9 25 •+ 3 9, were ma1nta1ned under 5tandard 1a60rat0ry c0nd1t10n5 at temperature 22 0 C •+ 2 ° C, re1at1ve hum1d1ty 50 •+ 10% and a 12-h ph0t0per10d. C0mmerc1a1 pe11et d1et (H1ndu5tan Lever, 1nd1a) and de10n15ed water were pr0v1ded ad 1161tum. Lead n1trate (P6(N03)2, m01. wt. 331.20, E. Merck, 1nd1a) and a1um1n1um 5u1phate (A12(504) 3 • 18H20, m01. wt. 666.42, 5ara6ha1 M. Chem1ca15, 1nd1a) were u5ed 51nce the5e 5a1t5 are kn0wn t0 exh161t d1fferent de9ree5 0f c1a5t09en1c1ty 1n mamma15 (Dh1r, 1985; R0y, 1987).

Exper1menta1pr0t0c01 7he exper1ment wa5 carr1ed 0ut 0n 3 5et5 0f an1ma15. 0 n e 5et wa5 adm1n15tered 0ra11y w1th Phy11anthu5 fru1t extract (PFE) at a rate 0f 685 m 9 / k 9 6.w., wh11e the 0ther 5et wa5 91ven AA at a rate 0f 16.66 m 9 / k 9 6.w. f0r 7 c0n5ecut1ve day5. 7he th1rd 5et wa5 ma1nta1ned a5 c0ntr01 w1th0ut any prev10u5 pr1m1n9.

0 n day 7, 1 h after pr1m1n9, a 5pec1f1c d05e 0f the meta1 wa5 1njected 1ntraper1t0nea11y (1.p.) t0 the an1ma15. 7hree d05e5 0f P6(N03)2, V12. 10, 20 and 40 m 9 / k 9 6.W., and 3 d05e5 0f A12(504) 3 • 18H20 , V12. 250, 500 and 1000 m 9 / k 9 6.W., Were U5ed. 7he 5a1t5 Were d15501Ved 1n n0rma1 5a11ne. A 51m11ar exper1ment Wa5 Carr1ed 0Ut W1th the n0npr1med c0ntr01 9r0up. Fr0m the pr1med 5et5, PFE-pr1med and AA-pr1med c0ntr015 were a150 51mu1tane0u51y ma1nta1ned. 5a11ne and m1t0myc1n C were 1njected 1.p. t0 an0ther 5et 0f n0n-pr1med m1ce t0 ma1nta1n ne9at1ve (veh1c1e) and p051t1ve (reference) c0ntr015, re5pect1ve1y. 51x an1ma15 were u5ed f0r every 5et 0f treatment/c0ntr01. An1ma15 were k111ed 24 h after adm1n15trat10n 0f the te5t chem1ca15. 90 m1n pr10r t0 death, each an1ma1 wa5 1njected 1.p. w1th 0.04% c01ch1c1ne (1 m1/100 9 6.w.). An1ma15 were k111ed 6y cerv1ca1 d1510cat10n, 60ne marr0w fr0m 60th femur5 wa5 rem0ved and f1u5hed 1n 0.075 M p0ta551um ch10r1de, 1ncu6ated at 37 ° C f0r 20 m1n, repe11eted and f1xed 1n c01d 1 : 3 91ac1a1 acet1c ac1d:ethan01. 511de5 were prepared 6y f1ume-dry1n9 and 5ta1ned 1n 61em5a (Pre5t0n et a1., 1987). 7he 511de5 were c0ded t0 e11m1nate 065erver 61a5. 7he m1t0t1c 1ndex wa5 determ1ned 6y c0unt1n9 the num6er 0f metapha5e5 1n 1000 ce115 per an1ma1, thu5 tak1n9 the c0unt fr0m a t0ta1 0f 6000 ce115 per 5et. 50 we11-5cattered metapha5e p1ate5 were 5canned fr0m each an1ma1, thu5 mak1n9 300 metapha5e5 per 5et. D1fferent type5 0f a6errat10n5 were rec0rded 5eparate1y. F0r ca1cu1at10n 0f 6reak5/ce11, chr0m050me and chr0mat1d 6reak5 were c0unted a5 1 6reak and rearran9ement5 a5 2 6reak5. 6ap5 were n0t 1nc1uded. 1n the c0unt 0f dama9ed ce115 (DC) a11 ce115 w1th at 1ea5t 1 a6errat10n (exc1ud1n9 9ap5) were 1nc1uded. 5tat15t1ca1 ana1y515 wa5 carr1ed 0ut f0110w1n9 the C0chran-Arm1ta9e trend te5t 1n each 1nd1v1dua1 chem1ca1 t0 5ee whether there wa5 any d05e-re1ated 1ncrea5e 1n a6errat10n5 (Mar9011n et a1., 1986). Further ana1y515 wa5 carr1ed 0ut f0110w1n9 1-way A N 0 V A (50ka1 and R0h1f, 1973) and Duncan•5 mu1t1p1e ran9e te5t t0 c0mpare the 519n1f1cance5 0f the d1fference5, 1f any, am0n9 d1fferent exper1menta1 5et5 (Harter, 1960; K0t2 and J0hn50n, 1982).

3

3 2

8"

1 2 1 -

2 1 -

1 2 1

2 3 4

3 2 2 4 4 2

M e a n 4- 5 E M

1 4 3 3 1 3

M e a n 4- 5 E M

2 -

1 3 3

M e a n 4- 5 E M

5 4 2 4 1 3

M e a n 4- 5 E M

1

-

M e a n 4- 5 E M

7

10 9

8•

N0n-pr1med

1 2 1 1 -

1 1 2

1 1 -

2 2 -

-

-

1

-

RR

P < 0.001

0.14 4- 0.01

0.12 0.20 0.14 0.14 0.14 0.10

0.11 4- 0.02

0.10 0.14 0.18

0.06 0.14 0.06

0.10 4- 0.01

0.10 0.08 0.08 0.16 0.12 0.06

0.02 4- 0.01

0.02 0.04 0.00 0.02 0.02

0.00

0.24 4- 0.01

0.24

0.26 0.22

12.33 4- 0.95

10 16 12 14 12 10

9.33 4-1.61

8 12 14

4 12 6

8.67 4- 0.84

10 8 8 12 8 6

1.67 4- 0.61

2 4 0 2 2

0

23.33 4- 0.94

22

26 22

DC(%)

CHR0M050ME5

CA/Ce11

MARR0W

2 1 3 1 1

2 3 2

2 5 2

2 3 1 3 2

1 1 1

2

8•

2 3 1 2

2 2

1 1

1 2 3 1 2

1 0 -

-

8••

1 2 1

1

2

1 1 -

-

-

RR

Phy11anthu5-pr1m¢d

0 F Mu5 mu5cu1u5

P < 0.001

0.08 4- 0.01

0.04 0.06 0.06 0.12 0.12 0.10

0.09 4- 0.02

0.04 0.10 0.12

0.04 0.12 0.14

0.08 4- 0.11

0.04 0.12 0.06 0.10 0.08 0.08

0.02 • 0.01

0.02 0.02 0.02 0.00 0.02

0.04

CA/Ce11

6.67 4- 0.84

4 6 6 10 6 8

8.33 4-1.41

4 10 10

4 12 10

7.33 4- 0.84

4 10 6 8 8 8

2.00 4- 0.52

2 2 2 0 2

4

DC(%)

8•, chr0mat1d 6 r e a k ; 8••, c h r 0 m 0 5 0 m e 6 r e a k ; R R , r e a r r a n 9 e m e n t 5 ; CA/Ce11, 6reak5/ce11; D C ( % ) , % 0f d a m a 9 e d ce115.

7 r e n d te5t P va1ue

40

20

10

0

3.5 m 9 / k 9 6.w.)

P051t1ve (reference) C0ntr01 (m1t0myc1n C,

( m 9 / k 9 6.w.)

C0ncentrat10n

EFFEC7 0F LEAD N17RA7E 0N 80NE

2 3 4 4 3 5

2

2

4 3 6 4 1 2

1 -

-

4

2

-

3

4

-

5

1 2 -

-

-

3 3 4 2 1 -

1 1

8"

1 1 1 1 -

8"

1 2 3 2

1

2

1

2

1

2

1 2 1

-

-

RR

A5c0r61c ac1d-pr1med

10.33 4- 0.95

0.13 4- 0.01

P < 0.001

0.19 4- 0.03

16.33 4- 2.03

14 16 20 22 8 18

12 10

0.16 0.12

0.16 0.20 0.20 0.28 0.08 0.22

8 10 8

14

4- 6.67 4-1.12

6 8 10 8 6 2

2.00 4- 0.52

0

2 4 2 2 2

DC(~)

0.10 0.12 0.10

0.18

0.08 4- 0.01

0.06 0.10 0.10 0.08 0.08 0.04

0.02 + 0.01

0.00

0.02 0.04 0.02 0.02 0.02

CA/Ce11

..j

3

7 r e n d te5t P va1ue

1000

500

250

0

3

-

1

2

-

1

1

2

1

1 1 1

5

5

3

3

1 1 -

5

4

4

4

5

Mean + 5EM

1 -

4

M e a n 5= 5 E M

1 1

3

7

M e a n 5= 5 E M

1 -

3

-

3

2

-

3 1

M e a n 5= 5 E M

-

-

M c a n 5= 5 E M

7

3.5 m 9 / k 9

6.w.)

9

2

10

C0ntr01 (m1t0myc1n C,

P051t1ve ( r e f e r e n c e )

6.w.)

8••

5ULPHA7E

8•

ALUM1N1UM

(m9/k9

0F

N0n-pr1med

2

C0ncentrat10n

EFFEC7

7A8LE

2

3

3

2

1

2

2

2

1

-

-

-

1

2

1

1

2

-

-

-

-

-

-

1

-

-

RR

0N

8

7

P < 0.001

12.83 5= 1.35

0.18

14

16

16

12

12

5= 0 . 0 1

0.18

0.22

0.22

0.16

0.14

0.18

12.00 5= 1.15

0.14

12

12

14

10

16

5= 0 . 0 2

0.16

0.16

0.16

0.10

0.16

0.08

7.33 + 0.42

0.10

8

8

8

8

6 6

5= 0.01

0.10

0.12

0.12

0.10

0.06 0.10

1.67 5= 0 . 6 1

0.02

2

2

0

4

2

0

5= 0 . 0 1

0.02

0.02

0.00

0.04

0.02

0.00

23.33 5= 0 . 9 4

5= 0.01

22

22

26

0.24

0.24

0.22

0.26

3

2

3

4

2

3

1

3

4

2

2

4

2

3

2

3

2 1

1

-

-

1

1

2

8•

-

-

1

-

2

1

2

-

1 1

1

2

-

1

-

-

1

-

-

-

-

2

-

-

-

-

-

-

RR

1

1

-

-

1

-

-

-

1

-

1

1

-

-

1

-

-

-

8••

P < 0.001

+ 0.02

0.11

0.14

0.08

0.16

0.08

0.06

0.12

5= 0.01

0.08

0.10

0.06

0.12

0.06

0.04

0.12

5= 0.01

0.07

0.04

0.08

0.04

0.08

0.04 0.12

5= 0 . 0 1

0.02

0.02

0.00

0.02

0.02

0.02

0.04

CA/Ce11

0 F Mu5 mu5cu1u5

Phy11anthu5-pr1med

CHR0M050ME5

DC(70)

MARR0W

CA/Ce11

80NE

-

0 2

0.00

0.02

0.02

0.08

0.10

3

1 1

-

1

1

2

0.08

0.12

0.12

0.18

P < 0.001

0.13

3

6 10

-

2

0.14

0.14

5= 0.01

5

2

2

1

1

9.00

2

8 12

1

-

-

1

0.14

5= 0 . 8 6

2

8

10

3

2

8

1

5= 0 . 3 3

9.67

10

10

10

8

10

10

5= 0.61

8.33

6

8

10

8

8 0.08

0.10

0.11

2

6

1

-

10

0.14

5= 0.01

4

10

-

2 1

7.00

4

6

1 -

5= 0 . 8 6

2 3

8 4

8.33 5= 1.21

0.11 + 0.01

6

8

6

8

14

6.00

2

0.08

0.10

0.06

0.12

0.16

+ 0.89

1

1

1

-

2

+ 0.52

2.00

0

2

2

2

4

8

1

4

-

1

-

1

-

-

-

-

0.02

2

DC(%)

0.12

2 2

3

8 8

1

8 4

6

4

2

-

1

-

-

0.02 0.04

0.02

1

2

-

-

CA/Ce11

+ 0.01

1

2

1

-

RR

2.00

1

8••

+ 0.52

1

2

8•

4

DC(%)

A5c0r61c ac1d-pr1med

00

309 Re5u1t5

Lead A d05e-dependent 1ncrea5e 1n chr0m050ma1 a6errat10n5 (CA) (7a61e 1) wa5 f0und 1n re5p0n5e t0 exp05ure t0 1ncrea51n9 d05e5 0f 1ead (P6). 7he re5p0n5e wa5 h19he5t 1n the 5et pr1med ear11er w1th AA, 1e55 1n the n0n-pr1med an1ma15 and 10we5t 1n the an1ma15 91ven P F E ear11er f0r 7 day5. Ana1y515 f0110w1n9 Duncan•5 mu1t1p1e ran9e te5t (7a61e 3) 5h0wed that at the 10we5t d05e 0f P6 91ven (10 m 9 / k 9 6.w.), pr1m1n9 w1th 60th A A and P F E decrea5ed the fre4uency 0f CA, th0u9h n0t t0 a 519n1f1cant 1eve1, a5 c0mpared t0 n0n-pr1med an1ma15 treated w1th P6. 1n an1ma15 91ven the h19he5t d05e 0f P6 (40 m 9 / k 9 6.w.), pr1m1n9 w1th

P F E decrea5ed, wh11e A A pr1m1n9 1ncrea5ed the fre4uency 0f CA a5 c0mpared t0 the n0n-pr1med 5et. 80th chan9e5 were at a 519n1f1cant 1eve1 ( P < 0.05). 7 h e fre4uency 0f d1v1d1n9 ce115 decrea5ed w1th 1ncrea51n9 d05e5 0f 1ead. 7h15 1nh161t0ry effect 1nduced 6y the 2 10wer d05e5 0f P6 (10 and 20 m 9 / k 9 6.w.) wa5 reduced 6y 60th P F E and AA. W1th the h19he5t d05e 0f 40 m 9 / k 9 6.w., PFE pr1m1n9 1ncrea5ed the fre4uency 0f d1v1d1n9 ce115, 6ut A A d1d n0t have any effect and the n0n-pr1med a5 we11 a5 AA-pr1med 5et5 5h0wed a1m05t 51m11ar m1t0t1c 1ndex va1ue5 (7a61e 4).

A1um1n1um A d05e-dependent 1ncrea5e 0ccurred 1n re-

7A8LE 3 MUL71PLE C0MPAR150N F0R 8REAK5/CELL 0rder1n9 the 5amp1emean5: D05e: 10 m9 P6(N03)2 Ne9 C A5c C 0.02 0.02

PhyUC 0.02

AP 10 na9 P6 0.08

PP 10 m9 P6 0.08

NP 10 na9 P6 0.10

PhyUC 0.02

PP 20 m9 P6 0.09

NP 20 m9 P6 0.11

AP 20 m9 P6 0.13

Phy11C 0.02

PP 40 m9 P6 0.08

NP 40 na9 P6 0.14

AP 40 m9 P6 0.19

PhyU C 0.02

PP 250 m9 A1 0.07

NP 250 m9 A1 0.10

AP 250 m9 A1 0.11

Phy11C 0.02

PP 500 na9 A1 0.08

AP 500 m9 A1 0.11

NP 500 m9 A1 0.14

Phy11C 0.02

PP 1000 m9 A1 0.11

AP 1000 m9 A1 0.13

NP 1000 m9 A1 0.18

D05e: 20 na9 P6(N03)2

Ne9 C 0.02

A5c C 0.02

D05e: 40 ra9 P6(N03)2

Ne9 C 0.02

A5c C 0.02

D05e: 250 na9 A12(504)3-18H20

Ne9 C 0.02

A5c C 0.02

D05e: 500 m9 A12(504)3-18H20

Ne9 C 0.02

A5c C 0.02

D05e: 1000 na9 A12(804)3-18H20

Ne9 C 0.02

A5c C 0.02

7he 610ck5wh1chare under11nedt09ether are n0t 519n1f1cant1yd1fferent (at 1eve15%) am0n9 each 0ther.

Ne9 C, 5a11ne;A5c C, a5c0r61cac1d-pr1med1njected w1th 5a11ne;Phy11C, Phy11anthu5-pr1med1njected w1th 5a11ne; PP, PhyUanthu5pr1med 1njected w1th chem1ca1;AP, a5c0r61cac1d-pr1med1njected w1th chem1ca1;NP, n0n-pr1med 1njected w1th chem1ca1.

310 5p0n5e t0 a1Um1n1Um (A1) treatment a5 5h0Wn 6y 519n1f1Cant P Va1Ue5 f0110W1n9 the trend te5t (7a61e 2). 7he mu1t1p1e c0mpar150n te5t 5h0wed (7a61e 3) that pr1m1n9 w1th 60th P F E and AA 5uppre55ed the fre4uency 0f CA c0mpared t0 the n0n-pr1med A1-treated 5et. 7he 0n1y except10n wa5 1n an1ma15 91ven the 10we5t meta1 d05e (250 m 9 / k 9 6.w.) after A A pr1m1n9. 7he decrea5e5 1nduced 6y PFE and A A were 519n1f1cant 1n an1ma15 91ven the 2 h19her d05e5 0f meta1 (500 m9 and 1000 m 9 / k 9 6.w.) 1n c0mpar150n w1th the n0n-pr1med 5et. 7he 2 pr1med 5et5 d1d n0t d1ffer 519n1f1cant1y fr0m each 0ther (7a61e 3). Ana1y515 f0110w1n9 Duncan•5 mu1t1p1e ran9e te5t 5h0wed that the decrea5e 1n the fre4uency 0f ce11 d1v1510n 1nduced 6y a11 3 d05e5 0f the meta1 wa5 a11ev1ated 6y 60th P F E and A A pr1m1n9, w1th the f0rmer m0re effect1ve than AA (7a61e 4).

D15cu5510n P6 15 kn0wn f0r 1t5 c1a5t09en1c act1v1t1e5 1n d1fferent te5t 5y5tem5, 1nc1ud1n9 p1ant5 11ke A111um 5at10um, Va1115ner1a 5p1ra115 (Dh1r et a1., 1986), 7rade5cant1a 5p. (Dryan0v5ka, 1987) and an1ma15 11ke m1ce (Dh1r et a1., 1985a) and m054u1t0 (5harrna et a1., 1988) and human 1euk0cyte5 (8eckman et a1., 1982). Var10u5 chem1Ca15 5uch a5 f1-a1an1ne (E1-Wa55ef, 1988), 1r0n (Dh1r et a1., 19856), 5e1en1um (Chakra60rty et a1., 1987) have 6een 5h0wn t0 ame110rate the t0x1c1ty 0f P6 e1ther 6y prec1p1tat1n9 the meta1 0r 6y re1ea51n9 60und P6 fr0m t155ue. A1, t0 a 1e55er extent, ha5 6een 5een t0 1nduce chr0m050ma1 a1terat10n5 1n te5t 5y5tem5 1n v1v0 and 1n v1tr0 (H0we115, 1984; R0y, 1987; R0y et a1., 1988).

7A8LE 4 MUL71PLE C0MPAR150N F0R M17071C 1NDEX 0rder1n9 the 5amp1emean5: D05e: 10 m9 P6(N03)2 PP 10 m9 P6 Phy11C 3.08 3.06

Ne9 C 3.04

A5c C 3.01

AP 10 na9 P6 2.97

NP 10 m9 P6 2.69

D05e: 20 m9 P6(N03) 2 Phy11C Ne9 C 3.06 3.04

A5c C 3.01

PP 20 na9 P6 2.96

AP 20 m9 P6 2.91

NP 20 m9 P6 1.94

D05e: 40 m9 P6(N03)2 PhyUC Ne9 C 3.06 3.04

A5c C 3.01

PP 40 m9 P6 2.59

AP 40 m9 P6 1.68

NP 40 na9 P6 1.39

D05e: 250 m9 A12(504)3.18H20 Phy11C AP 250 na9 A1 3.06 3.05

Ne9 C 3.04

PP 250 m9 A1 3.02

A5c C 3.01

NP 250 m9 A1 2.91

D05e: 500 m9 A12(504) 3" 18H20 Phy11C Ne9 C 3.06 3.04

A5c C 3.01

AP 500 m9 A1 2.93

PP 500 m9 A1 2.81

NP 500 m9 A1 2.40

D05e: 1000 m9 A12(504)3• 18H20 PhyUC Ne9 C 3.06 3.04

A5c C 3.01

PP 1000 m9 A1 2.65

AP 1000 na9 A1 2.30

NP 1000 na9 A1 1.29

7he 610ck5wh1chare under11ned t09ether are n0t 519n1f1canf1yd1fferent (at 1eve15%) am0n9 each 0ther. Ne9 C, 5a11ne;A5c C, a5c0r61c ac1d-pr1med1njected w1th 5a11ne; Phy11C, Phy11anthu5-pr1med1njected w1th 5a11ne; PP, Phy11anthu5pr1med 1njected w1th chem1ca1;AP, a5c0r61cac1d-pr1med1njected w1th chem1ca1;NP, n0n-pr1med 1njected w1th chem1ca1.

311 7he r01e 0f var10u5 p1ant extract5 a5 de5muta9en5 and ant1muta9en5 15 6e1n9 1ncrea51n91y rec09n15ed (5ee Dh1r, 1989). 7he5e extract5 act d1rect1y 0n muta9en5 0r 0n th05e a9ent5 that 1nh161t the 9enerat10n 0f act1ve muta9en1c f0rm5 (Kada et a1., 1985). Var10u5 p1ant extract5 have 5h0wn ant1muta9en1c and ant1carc1n09en1c pr0pert1e5 (Y05h1kawa et a1., 1981; AUdr1ck et a1., 1986; Fuj1m0t0 et a1., 1987; Ja1n et a1., 1987). U51n9 an exper1menta1 pr0t0c01 51m11ar t0 the pre5ent 0ne, 1n wh1ch m1ce were pr1med w1th carr0t and 5p1nach ju1ce, A6raham et a1. (1986) 5h0wed the 5uppre5510n 0f m1cr0nuc1e1 1nduced 6y cyc10ph05pham1de. A 1ar9e num6er 0f ve9eta61e ju1ce5 were a150 f0und t0 reduce CA 1n rat 60ne marr0w ce115 1nduced 6y d1methy16en2[a]anthracene (1t0 et a1., 1986). 7 h e effect5 0f the5e ve9eta61e ju1ce5 were attr16uted var10u51y t0 50me heatre515tant c0mp0und (Fuj1m0t0 et a1., 1987), v1tam1n C (8r1n, 1982), ch10r0phy111n ( 0 n 9 et a1., 1986), 8-car0tene (A6raham et a1., 1986) 0r t0 the 1nteract10n 6etween d1fferent c0mp0und5. 1n the pre5ent 5tudy, pr1m1n9 w1th P F E wa5 f0und t0 aff0rd a 5tr0n9er pr0tect10n a9a1n5t 60th chem1ca15 than pr1m1n9 w1th AA. 7he 1atter wa5 anta90n15t1c 0n1y a9a1n5t A1 and the 10we5t d05e 0f P6, 6ut wa5 519n1f1cant1y 5yner915t1c 1n an1ma15 91ven the h19he5t d05e 0f P6 (40 m 9 / k 9 6.w.). 7he 610ava11a6111ty 0f v1tam1n C ha5 6een rec0rded t0 6e h19h 1n fru1t extract 0f Phy11anthu5 em611ca; 10 m1 0f the extract (c0nta1n1n9 0n1y 8.7 m9) wa5 f0und t0 6e a5 effect1ve a5 100 m9 0f 5ynthet1c v1tam1n C a5 ce11n ta61et5 (R0y et a1., 1987). 7he 5yner915t1c effect 0f AA 1n an1ma15 adm1n15tered the 2 h19her c0ncentrat10n5 0f P6 re5em61e5 a 51m11ar effect 5een w1th mercury. 1n th15 ca5e a150, the 1ntake 0f h19h 1eve15 0f A A 1ncrea5ed the uptake and retent10n 0f the meta1 (5ee H0m19 and M05er, 1981). 5evera1 1n v1v0 and 1n v1tr0 5tud1e5 have a150 5h0wn that v1tam1n C enhanced the 9en0t0x1c1ty 0f certa1n chem1ca15 and c0mp1ex m1xture5 (Kr15hna et a1., 1986; 5ham6er9er, 1984; We1t6er9, 1987). 7he 9reater eff1cacy 0f PFE may 6e attr16uted t0 the pre5ence 0f 0ther natura1 1n9red1ent5 1n c0m61nat10n w1th AA. 7he 1nteract10n 0f the5e 1n9red1ent5 1n the fru1t extract w1th the t0x1c meta111c 5a1t5 15 m0re 5ucce55fu1 1n reduc-

1n9 the 9en0t0x1c effect5 than am0unt 0f 5ynthet1c AA.

an e4u1va1ent

Ackn0w1ed9ement5 7he auth0r5 are 9ratefu1 t0 Pr0fe550r A.K. 5harma, Pr09ramme C0-0rd1nat0r, f0r the fac111t1e5 pr0v1ded and t0 the Un1ver51ty 6rant5 C0mm15510n, New De1h1 and t0 the C0unc11 0f 5c1ent1f1c and 1ndu5tr1a1 Re5earch, New De1h1 f0r f1nanc1a1 a5515tance.

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