Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Effect of n-3 Fatty Acid Deficiency on Fatty Acid Composition in Brain, Retina and Liver Using a Novel Artificial Rearing System

인공 사육 동물 모델 시스템을 이용한 n-3 지방산 결핍이 쥐의 뇌, 망막, 간의 지방산 조성에 미치는 영향

  • Lim, Sun-Young (Division of Marine Environment & BioScience, Korea Maritime University)
  • 임선영 (한국해양대학교 해양환경생명과학부)
  • Published : 2005.04.01
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Abstract

Docosahexaenoic acid (22:6n-3, DHA) is highly enriched in membrane of brain and retina, and plays an important role in maintaining an optimal function of the central nervous system. We investigated the effect of n-3 fatty acid deficiency on rat brain, retina and liver fatty acyl composition at two different ages (3 wks and 15 wks) under DHA deficient condition. Rat pups born to dams fed a diet with $3.1\%$ of total fatty acids as $\alpha-linolenic$ acid (LNA) were fed using an artificial rearing system either an n-3 deficient (n-3 Def) or n-3 adequate (n-3 Adq) diet. Both diets contained $17.1\%$ linoleic acid (LA) but the n-3 Adq diet also contained $3.1\%$ LNA. Rats consuming the n-3 Def diet showed a lower brain $(50\%\;in\;13\;wks\;and\;70\%\;in\;15\;wks,\;p<0.05)$ and retinal $(50\%\;in\;13\;wks\;and\;63\%\;in\;15\;wks,\;p<0.05)$ DHA than those on the n-3 Adq diet, which was largely compensated for by an increase in docosapentaenoic acid (22:5n-6, DPAn-6). In the liver of the n-3 Def group, the percentage of DHA decreased by $97\%$ at 3 wks of age with an apparent increase in DPAn-6 relative to the n-3 Adq group (p<0.05), while there was a $65\%$ lower liver DHA in n-3 Def group at 15 wks of age than the n-3 Adq group (p<0.05). Liver arachidonic acid (20:4n-6, AA) was increased at 3 wks of age but decreased at 15 wks of age in the n-3 Def group compared with n-3 Adq group (p<0.05). In conclusion, the replacement of DHA by DPAn-6 in brain and retina fatty acid composition may be related to the suboptimal function in spatial learning, memory and visual acuity. This artificial rearing method presents a first generation model for n-3 deficiency that is similar to the case of human nutrition that commonly employed two generation model.

본 연구에서는 인공 사육 모델 시스템과 인공 쥐용 분유를 이용하여 출생 직후 12시간 이내에 엄마 쥐들로부터 신생 쥐를 분리하여 n-3 지방산 결핍분유와 n-3 지방산 적절분유로 3주 동안 사육한 후(이유기) 각각 장기들을 취하고 또한 성숙 쥐(15 wks)의 뇌, 망막, 간을 취하여 뇌 성장 발달 과정 동안 n-3지방산 결핍이 이들 장기들의 지방산조성 변화에 미치는 영향을 알아보고자 한다. 이유기와 성숙기 쥐의 뇌 지방산 조성은 n-3 지방산 적절군과 대조군과 비교했을 때, n-3지방산 결핍군의 경우 n-6지방산의 함량은 높았으나 (p<0.05), 총 monounsaturated 지방산과 총 n-3 지방산의 함량은 낮았고(p<0.05), 총 saturated 지방산의 경우는 변화가 없었다. 기대했듯이 뇌 DHA의 경우 n-3 지방산 결핍군은 n-3 지방산 적절군과 대조군에 비해 이유기 때 $50\%$, 성숙기에는 $70\%$까지 감소하였고 반면 n-6 계열인 DPAn-6의 함량은 상당히 증가하였음을 살펴볼 수가 있었다. 이유기의 망막의 총 saturated 지방산과 총 monounsaturated 지방산에는 식이군 사이에 변화가 없었고 22:4n-6, DPAn-6와 총 n-6 지방산의 함량은 n-3 지방산 결핍군에서 n-3 지방산 적절군과 대조군에 비해 증가하였으나(p<0.05), 22:5n-3, DHA와 총 n-3 지방산의 함량은 상당히 감소하였음을 관찰할 수가 있었다(p<0.05). 성숙 쥐 망막의 지방산조성의 경우 20:4n-6, 22:4n-6, 22:5n-6와 총 n-6 지방산의 함량은 n-3 지방산 결핍군에서 n-3 지방산 적절군과 대조군에 비해 증가하였으나(p<0.05), 22:5n-3, DHA와 총 n-3 지방산의 함량은 상당히 감소하였다(p<0.05). 특히 n-3 결핍군의 망막 DHA는 이유기 때 $57\%$ 감소에서 성숙기에 $63\%$까지 감소하였다. (p<0.05). 간의 경우, 이유기의 n-3 지방산 결핍군의 DHA는 $65\%$ 감소한 반면 간의 DPAn-6는 $59\%$ 증가하였다. 흥미로운 것은 이유기의 간 지방산 조성과는 대조적으로 성숙기의 n-3 지방산 결핍군의 18:2n-6, 20:3n-6, 20:4n-6와 22:4n-6 지방산 함량이 현저히 감소된 것과는 달리 DPAn-6의 함량이 n-3 지방산 적절군보다 $143\%$까지 증가되었음 관찰할 수 가 있었다(p<0.05). 이상의 장기들의 n-3지방산 결핍군은 이유기와 성숙기에서 DPAn-6/DHA비와 n-6/n-3비 또한 n-3 지방산 적절군과 대조군에 비해 컸으나(p<0.05) DPAn-6와 DHA의 합에는 변화가 없었다. 따라서 뇌 성숙 발달 과정동안 n-3 지방산 결핍은 뇌의 DHA의 결핍을 초래하고 이러한 비정상적인 조건에서 DPAn-6의 증가를 유발하였으며 이러한 지방산 조성의 변화와 공간과 후각에 기초를 하는 기억 학습 능력 저하의 관련성에 대해서는 앞으로 연구가 진행되어져야 할 것으로 사료되어진다.

Keywords

References

  1. Innis SM. 1991. Essential fatty acids in growth and development. Prog Lipid Res 30: 39-103 https://doi.org/10.1016/0163-7827(91)90006-Q
  2. Okuyama H, Kobayashi T, Watanabe S. 1996. Dietary fatty acids-The n-6/n-3 balance and chronic elderly diseases: Excess linoleic acid and relative n-3 deficiency syndrome seen in japan. Prog Lipid Res 35: 409-457 https://doi.org/10.1016/S0163-7827(96)00012-4
  3. Salem N. 1989. Omega-3 fatty acids: Molecular and biochemical aspects. In New protective roles of selected nutrients in human nutrition. Spiller G, Scala J, eds. Alan R. Liss, New York. p 109-228
  4. Tinoco J. 1982. Dietary requirements and functions of alpha-linolenic acid in animals. Prog Lipid Res 21: 1-45 https://doi.org/10.1016/0163-7827(82)90015-7
  5. Yavin E, Green P. 1998. Mechanisms of docosahexaenoic acid accretion in the fetal brain. J Neurosci Res 15: 129-136
  6. Bourre JM, Francois M, Youyou A, Dumont O, Piciotti M, Pascal G, Durand G. 1989. The effects of dietary alphalinolenic acid on the composition of nerve membranes, enzymatic activity, amplitude of electrophysiological parameters, resistance to poisons and performance of learning tasks in rats. J Nutr 119: 1880-1892
  7. Weisinger HS, Vingrys AJ, Bang VB, Sinclair AJ. 1999. Effects of dietary n-3 fatty acid deficiency and repletion in the guinea pig retina. Invest Ophthalmol Vis Sci 40: 327-338
  8. Weisinger HS, Vingrys AJ, Sinclair AJ, 1996. Effect of dietary n-3 deficiency on the electroretinogram in the guinea pig. Ann Nutr Meta 40: 91-98 https://doi.org/10.1159/000177900
  9. Birch EE, Hoffman DR, Uauy R, Birch DG, Prestidge C. 1998. Visual acuity and the essentiality of docosahexaenoic acid and arachidonic acid in the diet of term infants. Pediatr Res 44: 201-209 https://doi.org/10.1203/00006450-199808000-00011
  10. Neuringer M, Connor WE, Lin DS, Barstad L, Luck S. 1986. Biochemical and functional effects of prenatal and postnatal ${\omega}3$ fatty acid deficiency on retina and brain in rhesus monkeys. Proc Natl Acad Sci USA 83: 4021-4025
  11. Champoux M, Hibbeln JR, Shannon C, Majchrzak S, Suomi SJ, Salem N, Higley JD. 2002. Fatty acid formula supplementation and neuromotor development in rhesus monkey neonates. Pediatr Res 51: 273-281 https://doi.org/10.1203/00006450-200203000-00003
  12. Moriguchi T, Greiner RS, Salem N. 2000. Behavioral deficits associated with dietary induction of decrease brain docosahexaenoic acid concentration. J Neurochem 75: 2563-2573 https://doi.org/10.1046/j.1471-4159.2000.0752563.x
  13. Greiner RS, Moriguchi T, Slotnick BM, Hurron A, Salem N. 2001. Olfactory discrimination deficits in n-3 fatty aciddeficient rats. Physiol Behav 72: 379-385 https://doi.org/10.1016/S0031-9384(00)00437-6
  14. Catalan JN, Moriguchi T, Slotnick BM, Murthy M, Greiner RS, Salem N. 2002. Cognitive deficits in docosahexaenoic acid deficient rats. Behav Neurosci 116: 1022-1031 https://doi.org/10.1037/0735-7044.116.6.1022
  15. Moriguchi T, Salem N. 2003. The recovery of brain docosahexaenoate subsequent to dietary n-3 fatty acid insufficiency leads to recovery of spatial task performance. J Neurochem 87: 297-309 https://doi.org/10.1046/j.1471-4159.2003.01966.x
  16. Tinoco J, Babcock R, Hincenbergs I, Medwadowski B, Miljanich P. 1978. Linolenic acid deficiency: Changes in fatty acid patterns in female and male rats raised on a linolenic acids-deficient diet for two generations. Lipids 13: 6-17 https://doi.org/10.1007/BF02533360
  17. Ward G, Woods J, Reyzer M, Salem N. 1996. Artificial rearing of infant rats on milk formula deficient in the n-3 essential fatty acids: A rapid method for the production of experimental n-3 deficiency. Lipids 31: 71-77 https://doi.org/10.1007/BF02522414
  18. Ward GR, Huang YS, Xing HC, Bobik E, Wauben I, Auestad N, Montalto M, Wainwright PE. 1999. Effect of gammalinolenic acid and docosahexaenoic acid in formulae on brain fatty acid composition in artificially reared rats. Lipids 34: 1057-1063 https://doi.org/10.1007/s11745-999-0457-6
  19. Reeves PG, Neilsen FH, Fahey GC. 1993. Committee report on the AlN-93 purified rodent diet. J Nutr 123: 1939-1951
  20. Kanno T, Koyanagi N, Katoku Y, Yonekubo A, Yajima T, Kuwata T, Kitagawa H, Harada E. 1997. Simplified preparation of refined milk formula comparable to rat's milk: influence of the formula on development of the gut and brain in artificially reared rat pups. J Pediatr Gastroenterol Nutr 24: 244-252
  21. Hoshiba J. 1996. Automatic feeder for newborn rat use within 12 hours of birth. Contemporary Topics in Lab Anim Sci 35: 83-86
  22. Lim SY, Moriguchi T, Lefkowitz B, Loewke J, Majchrzak S, Hoshiba J, Salem N. 2003. Artificial feeding of an n-3 essential fatty acid-deficient diet leads to a loss of brain function in the first generation. In Essential fatty acids and eicosanoids. Huang YS, Lin SJ, Huang PC, eds. AOCS Press, Champaigan, IL. p 122-131
  23. Folch J, Lees M, Sloane-Stanley G. 1957. A simple method for the isolation and purification of total lipid from boron fluoride-methanol. J Biol Chem 226: 495-509
  24. Morrison WR, Smith LM. 1959. Preparation of fatty acid methyl esters and dimethylacetals from lipids with boronfluoride-methanol. J Lipid Res 5: 600-608
  25. Salem N, Reyzer M, Karanian J. 1996. Losses of arachidonic acid in rat liver after alcohol inhalation. Lipids 31: S153-156 https://doi.org/10.1007/BF02637068
  26. Salem N, Litman B, Kim H-Y, Gawrisch K. 2001. Mechanisms of action of docosahexaenoic acid in the nervous system. Lipids 36: 945-959 https://doi.org/10.1007/s11745-001-0805-6
  27. Gerbi A, Zerouga M, Maixent JM, Debray M, Durand G, Bourre JM. 1999. Diet deficient in alpha-linolenic acid alters fatty acid composition and enzymatic properties of Nat(+), K(+)- ATPase isoenzymes of brain membranes in the adult rat. J Nutr Biochem 10: 230-236 https://doi.org/10.1016/S0955-2863(99)00002-9
  28. Gerbi A, Zerouga M, Debray M, Durand G, Chanez C, Bourre JM. 1994. Effect of fish oil diet on fatty acid composition of phospholipids of brain membranes and on kinetic properties of Nat (+), K( +)- ATPase isoenzymes of weaned and adult rats. J Neurochem 62: 1560-1569 https://doi.org/10.1046/j.1471-4159.1994.62041560.x
  29. Niu SL, Mitchell DC, Lim SY, Wen ZM, Kim HY, Salem N, Litman BJ. 2004. Reduced G protein-coulped signaling efficiency in retinal rod outer segments in response to n-3 fatty acid deficiency. J Biol Biochem 279: 31098-31104
  30. Weisinger HS, Armitage JA, Jeffrey BG, Mitchell DC, Moriguchi T, Sinclair AJ, Weisinger RS, Salem N. 2002. Retinal sensitivity loss in third-generation n-3 PUFA-deficient rats. Lipids 37: 759-765 https://doi.org/10.1007/s11745-002-0958-3
  31. Pawlosky RJ, Denkins Y, Ward G, Salem N. 1997. Retinal and brain accretion of long-chain polyunsaturated fatty acids in developing felines: the effects of corn oil-based maternal diets. Am J Clin Nutr 65: 465-472
  32. Neuringer M, Anderson GJ, Connor WE. 1988. The essentiality of n-3 fatty acids for the development and function of the retina and brain. Ann Rev Nutr 8: 517-541 https://doi.org/10.1146/annurev.nu.08.070188.002505
  33. Moriguchi T, Lim SY, Greiner R, Lefkowitz B, Loewke J, Hoshiba J, Salem N. 2004. Effect of an n-3-deficient diet on brain, retina, liver fatty acyl composition in artificially reared rats. J Lipid Res 45: 1437-1445 https://doi.org/10.1194/jlr.M400087-JLR200

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