1997, 20 (4): 54 59 J ou rna l of N anj ing A g ricu ltu ra l U n iversity Ξ (, 210095) (Β2 ) : Β2, Β2 (P > 0. 05);, ( 15 Λmo lgl ) Β2 ; Tw een 20, ; (Υ) 013% Tw een 20 Β2 A ; 015 mo lgl Β2 (P < 0. 01) ; ; Β2 ; ; S85212 UPTAKE OF CAROTENO ID S BY INTESTINAL M UCOUS CELL S OF LAY ERS AND GOATS IN V ITRO L iu Q ing and Zhou Guanghong (Co llege of A n im al Science & T echno logy, N an jing A gric U n iv, N an jing 210095) ABSTRACT U p take of Β2caro tene and lutein by layers and goats w as studied in v itro. T he result show ed up take of Β2caro tene and lu tein w as concen tration2dependen t and a con stan t up take percen tage of Β2 caro tene w as observed. T h is suggested caro teno ids w ere up taken by passive diffusion m echanism. U p take of caro teno ids by cells of layer w as temperature2independent w h ile up take of lutein and Β2caro tene at h igh concentration by goats w as affected significantly by incubation temperature. Tw een 20 inh ibited up take of caro teno ids among w h ich lutein w as mo re heavily inh ibited than Β2caro tene. In addition inh ibition of Β2 caro tene and p romo tion of lutein by ph lo retin w ere first repo rted. Key words layers; goats; Β2caro tene; lutein; up take in v itro A, (xan thoph lls) (caro tenes), Β2 ; [ 1 3, ; ] ; Go rab Ho llander T y2 [ 4 8 czkow sk i Scita ], Β2, Ξ : 1996211208
4 : 55 1 111 11111 Β2 (, ) ( ) (ph lo retin) Sigm a (Tw een 20), L 2Α2 ( ), ( 292 ),V E, (, ), L 2 11112 56 58 20 kg, (, 3 m ) 38 112 11211 W eiser [ 9 ], (PBS) 10 m ggm l,,, 80% [ 7 11212 Scita ] [ 8 11213 ], 15 m in,,, 1 h, 1 000 g 5 m in, PBS 3 113 R itter [10 ], ; ; 114 x θ SD (n= 6) ; SA S t 2 211 Β- 1, 38 Β2, (P > 0. 05) ; ( 2),,,,,, ( 3), 212 Β- Β2 (1 20 Λm o lgl ) (015 15 Λm o lgl ) (P > 0. 05) ; 38, (0. 5 15 Λm o lgl ) Β2 ( 15 Λm o lgl ) 0 (P < 0. 01) (P < 0. 01)
56 20 Β2 Β2caro tene 1 Β- Table 1 Uptake of Β-carotene at differen t concen tration by isolated in testinal cells (Λg g) L ayer Goat 38 0 38 0 1 29. 0 0. 7 (10. 8) 34. 0 3. 7 - - 2 - - 11. 9 2. 4 (2. 2) 13. 5 5. 0 5 136. 0 0. 7 (10. 1) 112. 0 3. 7 38. 1 0. 0 (2. 8) 27. 0 0. 2 10 263. 5 12. 4 (9. 8) 253. 0 8. 1 59. 5 8. 2 (2. 2) 43. 6 9. 9 15 424. 8 35. 1 (10. 5) 392. 0 4. 0 82. 5 7. 3 A (2. 0) 48. 4 5. 5 B 20 530. 0 119. 6 (9. 8) 540. 9 0. 6 125. 4 3. 0 A (2. 3) 80. 2 11. 3 B : (% ) P < 0. 01; P < 0. 05, N o te: V alue in braket is the rate of up take (% ). difference betw een values of the sam e anim al in the sam e line w ith different cap ital letters is h igh ly significant, tho se w ith different sm all letters is signifi2 canṫ T he sam e as in T able 22T able 7. L utein 2 Table 2 Uptake of lute in at differen t concen tration by isolated in testinal cells (Λg g) L ayer Goat 38 0 38 0 0. 5 29. 3 1. 2 (20. 3) 26. 3 1. 8 11. 4 2. 9 A (7. 9) 4. 9 0. 0 B 1 34. 0 3. 5 (11. 8) 28. 1 0. 0 17. 9 1. 4 A (6. 2) 5. 7 1. 4 B 5 46. 1 3. 8 (3. 2) 41. 0 0. 0 31. 7 4. 3 A (2. 2) 17. 9 2. 8 B 15 87. 4 5. 4 (2. 0) 73. 0 8. 8 65. 6 10. 3 A (1. 5) 38. 0 3. 4 B Gluco se 3 Table 3 Uptake of glucose at differen t concen tration by isolated in testinal cells (Λg g) L ayer Goat 38 0 38 0 5 62. 5 8. 9 A (13. 2) 18. 2 4. 9 B 43. 0 0. 0 A (8. 7) 0. 0 B 10 - - 61. 4 0. 0 A (6. 2) 12. 3 8. 7 B 20 78. 7 8. 0 A (5. 2) 47. 4 3. 5 B 95. 1 7. 9 A (4. 8) 24. 6 0. 0 B 30 137. 8 11. 9 A (4. 8) 73. 2 2. 9 B - - 40 200. 6 18. 1 A (4. 1) 64. 9 10. 0 B 115. 1 15. 9 A (2. 9) 61. 4 17. 4 B 60 217. 5 36. 2 A (3. 8) 69. 1 8. 7 B 137. 0 16. 8 A (1. 5) 73. 4 8. 7 B 213 Tween 20 Β-, Tw een 20 Β2 ( 4, 5), Υ (Tw een 20) ( Υ= 0. 3% ), Β2 ; Υ(Tw een 20), Β2 ; Υ(Tw een 20) = 0. 3%,
4 : 57 Tw een 20, Υ(Tw een 20) = 0. 1% Υ= 1. 2% (g ), Tw een 20 Β2 2 Λm o lgl, g 518, 1, 5 Λm o lgl Β2 214 219 Β2 (2 Λm o lgl ), g 514 1139, (P < 0. 01) 4 Tween 20 Table 4 Effect of Tween 20 on uptake of caroteno ids by isolated in testinal cells from layer (Λg g) Caro teno ids Tw een 20 (Υ) 0. 1% 0. 3% 0. 6% 0. 9% 1. 2% 1 59. 2 4. 0 A 29. 0 0. 7 B 26. 5 7. 2 B 26. 7 1. 3 B 24. 8 2. 6 C (21. 9) (10. 8) (9. 9) (9. 9) (9. 2) 5 193. 1 22. 9 A 136. 0 7. 0 B 134. 3 7. 4 B 124. 0 3. 5 C 66. 1 2. 7 D (14. 3) (10. 1) (10) (9. 2) (4. 9) Β2 10 303. 0 29. 5 A 264. 0 12. 4 A 127. 9 8. 7 B 101. 6 2. 7 B 36. 2 0. 6 C Β2caro tene (11. 2) (9. 8) (4. 7) (3. 8) (1. 3) 15 401. 1 51. 9 A 425. 0 35. 2 A 145. 4 6. 1 B 111. 9 1. 3 B 42. 7 4. 8 C (9. 9) (10. 5) (3. 6) (2. 8) (1. 1) 20 464. 5 46. 3 B 530. 0 19. 6 A 155. 9 6. 6 C 130. 9 5. 8 C 46. 5 2. 6 D (8. 6) (9. 8) (2. 9) (2. 4) (0. 9) 2 89. 9 3. 7 A - 52. 7 2. 0 B 28. 1 5. 3 C 156. 0 3. 7 D L utein (15. 6) (6. 3) (4. 9) (2. 7) 5 Tween 20 Table 5 Effect of Tween 20 on uptake of caroteno ids by isolated in testinal cells from goat (Λg g) Caro teno ids Tw een 20 (Υ) 0. 1% 0. 3% 0. 6% 0. 9% 1. 2% 2 25. 4 2. 7 A 11. 9 2. 4 B 22. 2 1. 4 A 19. 8 1. 4 A 18. 3 1. 4 B Β2 (4. 7) (2. 2) (4. 1) (3. 7) (3. 4) Β2caro tene 10 47. 6 0. 00 A 59. 5 8. 2 A 19. 0 0. 00 B 17. 5 0. 1 B 11. 9 0. 2 B (1. 8) (2. 2) (0. 72) (0. 65) (0. 44) 2 47. 9 11. 5 A - 36. 2 2. 4 B 14. 6 0. 00 C 8. 9 3. 7 D L utein (8. 3) (6. 3) (2. 5) (1. 6) 214 Β-,, 015 mm o lgl (5, 40 mm o lgl ) (P < 0. 01 P < 0. 05),, 80 mm o lgl, (P > 0. 05) 015 mm o lgl Β2 (P < 0. 01 P < 0. 05), (P < 0. 01) ( 6, 7),
58 20 6 Table 6 Effect of phloretin on uptake of caroteno ids by isolated in testinal cells Carteno ids A nim al 5 gλg g - 1 Contro l Ph lo retin 0 mmo l L - 1 0. 5 mmo l L - 1 gλg g - 1 L ayer 46. 1 3. 8 A 3. 2 116. 8 15. 7 B 8. 4 L utein 5 Goat 31. 7 4. 3 A 2. 3 251 0. 1 B 18 Β2 5 L ayer 116 7. 0 A 8. 6 17. 9 0. 0 B 1. 3 Β2caro tene 5 Goat 38. 1 0. 0 a 2. 8 28. 6 0. 0 b 2. 2 10 L ayer 224 12. 4 A 8. 3 48. 4 4. 4 B 1. 7 7 Table 7 Effect of phloretin on uptake of glucose by isolated in testinal cells Gluco se gm g g - 1 Contro l Ph lo retin 0 mmo l L - 1 0. 5 mmo l L - 1 gm g g - 1 5 65. 2 8. 9 A 13. 2 9. 8 2. 4 B 2. 0 40 200. 6 18. 1 a 4. 1 134. 0 17. 8 b 3. 4 80 336. 4 119 4. 2 317. 4 20. 8 4. 0 3 311, Β2, 20 Λm o lgl,,, T yczkow sk i, [ 6 ] :, 15 Λm o lgl, T yzkow sk i Schaeffer [ 11 ], 312 Tween 20 Tw een 20,, [ 8, ], Tw een 20 3,,,, Yang [ 12 ],, Β2, A,
4 : 59 Tw een 20 Go rad Β2 A, Tw een 20 12 mm o lgl, V A Υ= 0. 3% Tw een 20, Β2, V A, 313,, Β2 5, Β2 Tw een 20,,,,, [ 13 ],, 1 Yang A, L arsen T W, T um e R K. Caro teno id and retino l concentration in serum, adipo se tissue and liver and caro teno id transpo rt in sheep, goats and cattle. A ust J A gric R es, 1992, 43: 1809 1817 2 Sch iedt K. N ew aspects of caro teno id m etabo lism in anim alṡ In: N I K rinsky ed. Caro teno id in Chem 2 istry and B io logy. N ew Yo rk: P lenum P ress, 1990. 247 267 3 K irton A H, C rane B, Paterson D J. Yellow fat in lam bs caused by caro teno ids p igm entation. N Z J A 2 gri R es, 1975, 18: 267 272 4 E l2go rab M I, U nderw ood B A, L oerch J D. T he ro les of bile salts in the up take of beta2caro tene and retino l by rat everted gut sacṡ B iochem B iophy A cta, 1975, 401: 265 277 5 Ho llander D, R uble P E. Beta2Caro tene intestinal abso rp tion: bile, fatty acid, ph, and flow rate ef2 fects on transpo rṫ Am J Physio l, 1978, 235(6): E686 691 6 T yczkow sk i J K, H am ilton P B. L utein as a model dihydroxycaro teno id fo r the study of p igm entation in ch ickenṡ Poultry Science, 1986, 65: 1141 1145 7 Scita G, A ponte G W, W o lf G. U p take and cleavage of beta2caro tene by cultures of rat sm all in testi2 nal cells and hum an lung fibroblastṡ In: L Packer ed. M ethods in Enzymo logy. San D iego: A cadem ic P ress, 1993. 213: 212 32 8, T um e R K, L arsen T W. Β2 1, 1996, 8 (4): 15 18 9 W eiser M M. Intestine ep ithelial cell surface m em brane glycop ro tein synthesiṡ J B io l Chem, 1973, 248: 2536 2541 10 R itter E D, Purcell A E. Caro teno id analytical m ethodṡ In. J C Bauernfeind ed. Caro teno ids as Co l2 o rants & V itam in A P recusso rṡ N ew Yo rk: A cadem ic P ress, 1981, 816 877 11 Schaeffer J. T yczkow sk i J K. Parkhurst C. Caro teno id compo sition of serum and egg yo lk s of hens fed diets varying in caro teno id compo sition. Poultry Science, 1988, 67: 608 614 12 Yang A, T um e R K. A com parison of beta2caro tene2sp litting activity iso lated from in testinalm uco sa of pasture2grazed sheep, goats and cattle. B iochem and M o lecular B io logy International, 1993, 30 (2): 209 217 13 1 1 :, 1985. 90 91 ( )