Acute Oral Toxicity and Anti-obesity Effect of Diglyceride Preparation Containing Conjugated Linoleic Acid in Rat

공액리놀레산 함유 디글리세라이드 식용유지 조성물의 rat에 대한 단회 경구투여독성 및 항비만 효과

  • Published : 2009.06.30

Abstract

The present study was carried out to investigate the acute oral toxicity and anti-obesity effects of a diglyceride preparation containing conjugated linoleic acid (DG+CLA). To test its acute oral toxicity, the DG+CLA was injected into 30 rats (15 males and 15 females) at dosage of 2,000 mg/kg and 5,000 mg/kg. Mortality rates, clinical signs, and body weight changes were monitored for 14 days following administration. According to the results, the lethal dose ($LD_50$) of DG+CLA was determined as >5,000 mg/kg in both sexes. There were no significant changes in general conditions, clinical signs, body weight, and gross lesions between the vehicle control and DG+CLA groups. For the anti-obesity studies, obese Zucker rats were randomly divided into 4 groups and fed saline, soybean oil, diglyceride, and DG+CLA, respectively, for 8 weeks. The DG+CLA groups presented significant differences in body weight, food efficiency ratio, serum lipid levels, and fat weight. Overall, the results showed that the DG+CLA did not have acute oral toxicity and reduced body weight, serum lipid levels, and fat gain.

단회 경구 투여 독성 평가를 위해 암 수 rat을 이용하여 시험물질 2,000 mg/kg과 5,000 mg/kg을 투여한 후 14일 동안 관찰한 결과, 투여에 기인한 사망례 및 체중변화, 특이한 일반증상은 관찰되지 않았다. 암컷 대조군 및 시험물질 2,000 mg/kg 투여군에서 투여 1일에 연변외 점액변이 관찰되었으나, 이는 투여에 기인한 일시적인 변화로 사료된다. 투여 2일부터는 암컷 대조군과 시험물질 투여군에서 특이한 일반증상이 관찰되지 않았다. 부검 시 암컷 대조군 및 암 수 시험물질 투여군에서 투여에 기인한 것으로 판단되는 육안소견은 관찰되지 않았다. 수컷 대조군 1례의 부검결과, 부고환, 정낭 및 양측 고환에서 소형 소견 관찰되어, 조직병리학적 검사를 실시한 결과, 부고환, 정낭 및 고환의 위축 소견이 관찰되었으나, 이는 자연 발생적인 소견으로 사료된다. 이상으로 공액리놀렌산이 함유된 디글리세라이드 식용유지 조성물을 rat에 단회 경구투여한 결과, $LD_50$값이 rat kg당 5,000 mg 이상으로 판단되며, 단회 경구 투여에 대한 독성을 나타내지 않는 것으로 판단된다. 항비만 효과를 조사하기 위해 obese Zucker rat에 saline, soybean oil, DG, DG+CLA를 8주간 투여한 후 효과를 비교하였다. 체중변화의 경우 DG+CLA 투여군은 대조군에 비하여 투여 후 3주째부터 통계학적으로 유의성 있게 체중 감소를 나타냈다. 1일 식이효율측정의 결과, soybean oil, DG 및 DG+CLA 투여군이 대조군에 비하여 통계학적으로 유의성 있는 1일 식이 효율의 증가를 나타냈다. 혈중지질분석의 결과, 총콜레스테롤 수치에서 soybean oil, DG 및 DG+CLA 투여군 모두 대조군에 비하여 통계학적으로 유의성 있게 총콜레스테롤 함량이 감소되었으며, 중성지방 수치는 soybean oil 및 DG 투여군에서만 대조군에 비하여 통계학적으로 유의성 있게 감소되었다. 지방조직무게 측정 결과, DG+CLA 투여군은 신장주위지방무게를 제외한 부고 환주위지방, 갈색지방 및 내장지방무게에서 대조군과 비교하여 통계학적으로 유의성 있게 지방조직무게가 감소되었다. 이상의 결과로 obese Zucker rat에 대한 5 mL/kg의 DG+CLA 경구투여는 CLA의 주요 이성질체인 c9, t11-CLA와 t10, c12-CLA가 hormone sensitive lipase와 carnitine palmitoyl transferase의 활성을 증가시켜 지방 분해를 촉진시켜 체내 지방의 축적을 막으며, 중성지방 콜레스테롤과 같은 지질 대사에 영향을 주어 항비만 효과를 나타내는 것으로 사료되어진다.

Keywords

References

  1. Kelly GS. Conjugated linoleic acid: A review. Altern. Med. Rev. 6: 367-384 (2001)
  2. Nakamura MT, Nara TY. Structure, function, and dietary regulation of ${\Delta}6$, ${\Delta}5$, and ${\Delta}9$ desaturases. Annu. Rev. Nutr. 24: 345-376 (2004) https://doi.org/10.1146/annurev.nutr.24.121803.063211
  3. Joseph AS, Miller GD. Potential health benefits of conjugated linoleic acid. J. Nutr. 19: 470S-471S (2000)
  4. Lin TY. Conjugated linoleic acid concentration as affected by lactic cultures and added linoleic acid. Food Chem. 67: 1-3 (1999) https://doi.org/10.1016/S0308-8146(99)00077-1
  5. Ha YL, Grimm NK, Pariza MW. Anticarcinogens from fried ground beer: Heat-altered derivatives of linoleic acid. Carcinogenesis 8: 1881-1887 (1987) https://doi.org/10.1093/carcin/8.12.1881
  6. Gniwotta C, Morrow JD, Roberts LJ, Kuhn H. Prostaglandin F2- like compounds, F2-isoprostanes, are present in increased amounts in human atherosclerotic lesions. Arterioscl. Throm. Vas. 17: 3236-3241 (1997) https://doi.org/10.1161/01.ATV.17.11.3236
  7. Pariza MW, Ha YL, Benjamin H, Sword JT, Gruter A, Chin SF, Storkson J, Faith N, Albright K. Formation and action of anticarcinogenic fatty acids. Adv. Exp. Med. Biol. 289: 269-272 (1991)
  8. Macdonald HB. Conjugated linoleic acid and disease prevention: A review of current knowledge. J. Nutr. 19: 111S-118S (2000)
  9. Sugano M, Tsujita A, Yamasaki M, Yamada K, Ikeda I, Kritchevsky D. Lymphaticrecovery, tissue distribution and metabolic effects of conjugated linoleic acid in rats. J. Nutr. Biochem. 8: 38-43 (1997) https://doi.org/10.1016/S0955-2863(96)00150-7
  10. Yurawecz MP, Hood JK, Pariza MW. Conjugated linoleic acid and atherosclerosis in rabbits. Atherosclerosis 108: 19-25 (1995) https://doi.org/10.1016/0021-9150(94)90034-5
  11. Terpstra AH, Javadi M, Beynen AC, Kocsis S, Lankhorst AE, Lemmens AG, Mohede IC. Dietary conjugated linoleic acids as free fatty acids and triacyglycerols similarly affect body composition and energy balance in mice. J. Nutr. 133: 3181-3186 (2003)
  12. Yamasaki M, Mansho K, Mishima H, Kasai M, Sugano M, Tachibana H, Yamada K. Dietary effect of conjugated linoleic acid on lipid levels in white adipose tissue of sprague-dawley rats. Biosci. Biotechnol. Biochem. 63: 1104-1106 (1999) https://doi.org/10.1271/bbb.63.1104
  13. Gaullier JM, Halse J, Hφye K, Kristiansen K, Fagertun H, Vik H, Gudmundsen O. Supplementation with conjugated linoleic acid for 24 months is well tolerated by and reduces body fat mass in healthy, overweight humans. J. Nutr. 135: 778-784 (2005)
  14. Riserus U, Berglund L, Vessby B. Conjugated linoleic acid (CLA) reduced abdominal adipose tissue in obese middle-aged men with signs of the metabolic syndrom: A randomised controlled trial. Int. J. Obesity 25: 1129-1135 (2001) https://doi.org/10.1038/sj.ijo.0801659
  15. Gaullier JM, Halse J, Hφivik HO, Hoye K, Syvertsen C, Nurminiemi M, Hassfeld C, Einerhand A, O'Shea M, Gudmundsen O. Six months supplementation with conjugated linoleic acid induces regional-specific fat mass decreases in overweight and obese. Brit. J. Nutr. 97: 550-560 (2007) https://doi.org/10.1017/S0007114507381324
  16. Kamphuis MM, Lejeune MP, Saris WH, Westerterp-Plantenga MS. The effect of conjugated linoleic acid supplementation after weight loss on body weight regain, body composition, and resting metabolic rate in overweight subjects. Int. J. Obesity 27: 840-847 (2003) https://doi.org/10.1038/sj.ijo.0802304
  17. Smedman A, Vessby B. Conjugated linoleic acid supplementation in humans-metabolic effects. Lipids 36: 773-781 (2001) https://doi.org/10.1007/s11745-001-0784-7
  18. Thom E, Wadstein J, Gudmundsen O. Conjugated linoleic acid reduces body fat in healthy exercising humans. J. Int. Med. Res. 29: 392-396 (2001) https://doi.org/10.1177/147323000102900503
  19. Mougios V, Matsakas A, Petridou A, Ring S, Sagredos A, Melissopoulou A, Tsigilis N, Nikolaidis M. Effect of supplementation with conjugted linoleic acid on human serum lipids and body fat. J. Nutr. Biochem. 12: 585-594 (2001) https://doi.org/10.1016/S0955-2863(01)00177-2
  20. Nagao K, Wang YM, Inoue N, Han SY, Buang Y, Noda T, Kouda N, OKamatsu H, Yanagita T. The 10-trans, 12-cis isomer of conjugated linoleic acid promotes energy metabolism in OLETF rats. Nutrition 19: 652-656 (2003) https://doi.org/10.1016/S0899-9007(03)00060-1
  21. Scimeca JA. Toxicological evaluation of dietary conjugated linoleic acid in male fischer 344 Rats. Food Chem. Toxicol. 36: 391-395 (1998) https://doi.org/10.1016/S0278-6915(97)00169-5
  22. O'Hagan S, Menzel A. A subchronic 90-day oral rat toxicity study and in vitro genotoxicity studies with a conjugated linoleic acid product. Food Chem. Toxicol. 41: 1749-1760 (2003) https://doi.org/10.1016/S0278-6915(03)00203-5
  23. Yang M, Cook ME. Dietary conjugated linoleic acid decreased cachexia, macrophage tumor necrosis factor-a production, and modifies splenocyte cytokines production. Exp. Biol. Med. 228: 51-58 (2003)
  24. Bontempo V, Sciannimanico D, Pastorelli G, Rossi R, Rosi F, Corino C. Dietary conjugated linoleic acid positively affects immunologic variables in lactating sows and piglets. J. Nutr. 134: 817-824 (2004)
  25. Soni MG, Kimura H, Burdock GA. Chronic study of diacylglycerol oil in rats. Food Chem. Toxicol. 39: 317-329 (2001) https://doi.org/10.1016/S0278-6915(00)00140-X
  26. Chengelis CP, Kirkpatrick JB, Marit GB, Morita O, Tamaki Y, Suzuki H. A chronic dietary toxicity study of DAG (diacylglycerol) in Beagle dogs. Food Chem. Toxicol. 44: 81-97 (2006) https://doi.org/10.1016/j.fct.2005.06.005
  27. Kasamatsu T, Ogura R, Ikeda N, Morita O, Saigo K, Watabe H, Saito Y, Suzuki H. Genotoxicity studies on dietary diacylglycerol (DAG) oil. Food Chem. Toxicol. 43: 253-260 (2005) https://doi.org/10.1016/j.fct.2004.10.001
  28. Takei A, Toi T, Takahashi H, Takeda Y, Moriwaki J, Takase H, Katsuragi Y. Effects of diacylgcerol-containing mayonnaise on lipid metabolism and body fat in human. Jpn. Nutr. Food 4: 89-94 (2001)
  29. Watanabe H, Tokimitsu I. Digestion and absorption of diacylglycerol. p.30. In: Diacylglycerol Oil, Katsuragi Y, Yasukawa T, Matsuo N, Flickinger BD, Toimitsu I, Matlok MG (eds). American Oil Chemistsi Society, Champaign, IL, USA (2004)
  30. Matsuyama T, Shoji K, Watanabe H, Shimizu M, Saotome Y, Nagao T, Matsuo N, Hase T, Tokimitsu I, Nakaya N. Effects of diacylglycerol oil on adiposity in obese children: Initial communication. J. Pediatr. Endoc. Met. 19: 795-804 (2006)
  31. Teramoto T, Watanabe H, Ito K, Omata Y, Furukawa T, Shimoda K, Hoshino M, Nagao T, Naito S. Significant effects of diacylglycerol on body fat and lipid metabolism in patients on hemodialysis. Clin. Nutr. 23: 1122-1126 (2004) https://doi.org/10.1016/j.clnu.2004.02.005
  32. Maki KC, Davidson MH, Tsushima R, Matsuo N, Tokimitsu I, Umporowicz DM, Dicklin MR, Foster GS, Ingram KA, Anderson BD, Frost SD, Bell M. Consumption of diacylglycerol oil as part of a reduced-energy diet enhances loss of body weight and fat in comparison with consumption of a triacylglycerol control oil. Am. J. Clin. Nutr. 76: 1230-1236 (2002)
  33. Nagao T, Watanabe H, Goto N, Onizawa K, Taguchi H, Matsuo N, Yasukawa T, Tsushima R, Shimasaki H, Itakura H. Dietary diacylglycerol suppresses accumulation of body fat compared to triacylglycerol in men in a double-blind controlled trial. J. Nutr. 130: 792-797 (2002)
  34. Yasunaga K, Glinsmann WH, Seo Y, Katsuragi Y, Kobayashi S, Flickinger B, Kennepohl E, Yasukawa T, Borzelleca JF. Safety aspects regarding the consumption of high-dose dietary diacylglycerol oil in men and women in a double-blind controlled trial in comparison with consumption of a triacylglycerol control oil. Food Chem. Toxicol. 42: 1419-1429 (2004) https://doi.org/10.1016/j.fct.2004.04.003
  35. Moon HS, Lee HG, Seo JH, Chung CS, Kim TG, Kim IY, Lim KW, Seo SJ, Choi YJ, Cho CS. Down-regulation of PPARgamma2- induced adipogenesis by PEGylated conjugated linoleic acid as the pro-drug: Attenuation of lipid accumulation and reduction of apoptosis. Arch. Biochem. Biophys. 456:19-29 (2006) https://doi.org/10.1016/j.abb.2006.10.002
  36. Normen L, Dutta P, Lia A, Andersson H. Soy sterol esters and beta-sitostanol ester as inhibitors of cholesterol absorption in human small bowel. Am. J. Clin. Nutr. 71: 908-913 (2000)
  37. Becker M, Staab D, von Bergman K. Long-term treatment of severe familial hypercholesterolemia in childhood with sitosterol and sitostanol. J. Pediatr. 122: 292-296 (1993) https://doi.org/10.1016/S0022-3476(06)80136-8
  38. Watanabe H, Onizawa K, Taguchi H, Kobori M, Chiba H, Naito S, Matsuo N, Yasukawa T, Hattori M, Shimasaki H. Nutritional characterization of diacylglycerols in rats. J. Jpn. Oil Chem. 46: 301-307(1997) https://doi.org/10.5650/jos1996.46.301
  39. Yasukawa T, Yasunaga K. Nutritional functions of dietary diacylglycerols. J. Oleo Sci. 50: 427-432 (2001) https://doi.org/10.5650/jos.50.427