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J. Korean Soc. Appl. Biol. Chem. 51(4), 288-293 (2008) Articles vitamin E Á» wp z 1 Áš 1 Á 1,2, * 1 w w œw, 2 w œw Anti-stress Effect during Long and Short-Term of Vitamin E in Mice 1 Seung Jin Lee 1, Min Seok Go 1, and Sang Mo Kang 1,2, * Department of Bioengineering at the Postgraduate School, Konkuk University, Seoul 143-701, Korea 2 Department of Microbial Engineering, Konkuk University, Seoul 143-701, Korea Received September 9, 2008; Accepted October 30, 2008 This study was to evaluate the protective effect of vitamin E against long and short-term stress in ICR mice. Two groups which had been bred for 5 months (equivalent to human beings aged 20) were treated by immobilization stress for 8 weeks with or without vitamin E, and one out of two groups was continuously bred until they become 18 months old (equivalent to human senescence) with or without Vitamin E. Afterwards, the changes of serum and hepatic metabolites were investigated on the basis of the index of stress-related in vivo oxidative damage. As a result, it was found that stress increases serum triacylglycerol and aspartate aminotransferase (AST) in the long and short-term, and decreases serum HDL-cholesterol. In addition, stress concerned the decrease of total antioxidant status (TAS) and superoxide dismutase (SOD) as well as the increase of malondialdehyde (MDA) in liver. These results suggest that stress in one s youth causes negative results in TG, HDL-cholesterol, AST, TAS, SOD and MDA measured in one s senescent. The administration of vitamin E in the stressed mice decreases serum TG and AST that are increased by stress, and exerts influence on the increase of serum HDL-cholesterol. Also vitamin E recovered the values of liver TAS, SOD and MDA in the stressed mice. In conclusion, vitamin E represented protective effect in the stressed mice to some degree. Key words: liver tissue, serum, stress, vitamin E y w,,, ¼ w p(stressor)š w, p y» w e w x p(stress)š w. p w w w v (hypothalamic-pituitaryadrenocortical. HPA) (sympathetic-adrenal medullary. SAM) yyg ƒƒ glucocorticoids catecholamines ƒk. Catecholamines w k 1, superoxide radical(o 2 ), hydrogen peroxide (H 2 O 2 ) hydroxyl radical(oh ) y (reactive oxygen species, ROS). *Corresponding author Phone: +82-2-450-3524, Fax: +82-2-3437-8360 E-mail: kangsm@konkuk.ac.kr doi:10.3839/jksabc.2008.050 ù w ROS w z wy z superoxide dismutase(sod), catalase(cat), gultathione peroxidase(gsh-px) ƒš. 4) ù ƒ» š wy» pw y ƒ» w» ƒ wy w, t wy vitamin E(α-tocopherol). 5,6) Vitamin E x ü w w yw y w wy, vitamin Eƒ wy y j peroxy-radical sw vitamin E radical xw ƒ y w». Vitamin E sü w, y p w s ü y w w w. p 7) vitamin E y p j w» ƒš vitamin E radical vitamin C glutathione y w vitamin E y. 8) p w y vitamin E 288

ƒ z z wy, ¾ p z ü vitamin Eƒ x wyz y e w w š ù, 5-8) p z ùkù» y w vitamin E ü z w ƒ. ICR ù 20 w 5 ¾ w, 8 p ƒw z ù» w 18 ¾ w 9) p w ü y t y ù 20 pƒ» 60 x y vitamin E y z rš wš, w 20, 9) 80 ƒw. x ü x. x z 4 ICR f Japan SLC, Inc.l x y ( 22±2 o C, 45%) w. šx 1 k z ù w 5 w 8 p ƒw 15 4 w, 5 ¾ w 8 p ƒw z 18 w 20 4 ù. w vitamin E α- tocopherol acetate Sigma(St. Louis, Missouri, USA)l œ, x α-tocopherol acetate 5g/50g w 0.4±0.6 g w w. p. p 7 w (C-7) 5 w 8 p ƒw p x (S-7), p vitamin E w (E-7) 8 p ƒw vitamin E w p x (S+E-7), 18 p w (C-18), 5 w 8 p z 18 p w x (S-18), 18 p vitamin E w (E-18), 5 w 8 p z 18 p vitamin E w x (S+E-18) w. x p immobilization stress(y p) w. p w y yw»( k) w yj p ƒwš w. p w vx p case (10 cm 25 cm) š ã k 40 g. 10 p w z x» w p z yw. x. x x w» w x 12 z diethyl ether w x w. w x 1 ew z 3,000 rpm 20 w x wš, vitamin E Á» wp z 289 ¾ 80 o C þ w, x z w. x. x triacylglycerol, HDL-cholesterol, aspartate aminotransferase(ast) d kit(bayer, NY, USA) w» ADVIA 1650(Bayer, USA) w w.. ü total antioxidant status(tas) TAS kit(randox Laboratories Ltd, Ardmore, UK) w Hitachi 7150(Hitachi, Japan) w. SODd SOD kit(cayman, USA) w ElA reader(molecular device, USA) w. Malondialdehyde(MDA) d BIOXTECH LPO-586 assay kit w dw. m. xw m SPSS program w ƒ x s³ t r ùkþ, 7 18 y t (paired ) wš, ƒ p<0.05 One-way ANOVA test ùkü, Duncan's multiple range test w. š x TG w y. 5 8 p ƒw z, y 18 z x TG w dw. Table 1. 7 p S-7 474.92 mg/dl, C-7 w 127% ƒw, p ƒw vitamin E n w S+E-7 381 mg/dl p w 19% w. w 7 18 p S- 7 474.92 mg/dl, S-18 537.08 mg/dl(t= 5.27, p ƒw vitamin E nw S+E-7 381 mg/dl, S+E-18 434.31 mg/dl(t= 3.645). x TG w p w ƒ Kissebah 1 w ùkü, Howard 1 š pƒ sü w s(macrophage) w TG w ƒ. Vitamin E n p w TG w û ùkù vitamin E ƒ ü y w» yk y yj»w. 1 w vitamin Eƒ p w TG zj z Gittleman 14) w ew. w ù 20 ww 7 p S-7 p ƒw vitamin E nw S+E-7 ù 60» ww 18 p S-18 p ƒw vitamin E n w S+E-18 TG w pƒ» TG w. mw p TG w ƒjš, vitamin E TG zw

290 ÁšÁ Table 1. The serum triglyceride of ICR mice that were treated by immobilization stress with or without vitamin E C, 208.42±18.12 c, 211.00±21.28 c -.365 E 211.55±22.73 c 219.91±28.26 c -.842 S+E 381.00±54.12 b 434.31±78.93 b -3.645**, S 474.92±30.84 a 537.08±42.08 a -5.273*** **: p<0.01, ***: p<0.001. w p ù» TG ƒj w q. x HDL-cholesterol w y. TG w x HDL-cholesterol w dw. Table 2. 7 p S-7 43.26 mg/ dl 17% w, p ƒ w vitamin E nw S+E-7 70.11 mg/dl p 62% ƒw. 7 18 p S-7 43.26 mg/dl, S-18 39.84 mg/dl (t=15.846) S-18 7.9% w š, C-7, C-18 ƒ, S+E-7 S+E-18 ƒ. w vitamin E nw E-7 E-18 C-7. x HDL-cholesterol ƒ p w w ùkù Lehtonen w 15). Howard p 1 w sƒ y LDL-cholesterol w w s ü y gl ƒ HDL-cholesterol g ƒ. w Table ù 7 18 LDL-cholesterol x dw C-7 25.14 mg/dl, E-7 23.25 mg/dl, S+E-7 40.12 mg/dl, S-7 43.18 mg/dl, E-7 p S-7 46% w, 18 vitamin E n w w. w vitamin Eƒ gl LDL-cholesterol x k, vitamin Eƒ x y w y w. 16) p ù» ww 18 ƒ ù 20 ww p HDLcholesterol ƒ w mw, pƒ TG w» HDLcholesterol w e. w 18 w p ƒw vitamin E nw S+E- 18 p S-18 w HDL-cholesterolƒ Table 2. The serum HDL-cholesterol of ICR mice that were treated by immobilization stress with or without vitamin E C 52.16±6.12 b, 54.83±3.19 c.-0.737 E 73.24±6.03 a 64.12±4.10 b 05.948*, S+E 70.11±4.98 a 69.21±4.11 a 01.836 S 43.26±3.74 c 39.84±2.96 d 15.846** *: p<0.05, **: p<0.01 Table 3. The serum aspartate aminotransferase of ICR mice that were treated by immobilization stress with or without vitamin E C 29.08±3.82 c, 31.75±4.03 c -1.434 E 29.73±5.24 c 32.36±2.94 c -1.256 S+E 43.77±8.22 b 43.08±5.66 b -0.337 S 49.67±6.69 a 54.42±3.37 a -2.077 73% ƒw mw, vitamin Eƒ HDLcholesterol ƒ z ƒ. Vitamin E nw E-18 C-18 HDLcholesterol ƒ 16% ùkù pƒ vitamin E w j z HDL-cholesterol w e ùkû. p vitamin E n z vitamin Eƒ HDL-cholesterol w e m w. x AST w y. TG w HDL-cholesterol w xl t AST w 17) dw. Table 3. 7 p S-7 49.67 µ/l C-7 w 70% ƒw, p ƒw vitamin E nw S+E-7 43.77 µ/l p S-7 w 11% w. p w AST wƒ p ü y x ASTƒ ƒw Woo 18) AST ƒ» x z p ù shock ƒwš w John 19) w. 7 18 p S-7 ù S-18 TGù HDL ùkù. w z p 2 w sƒ q z p q. 7 18 vitamin E n p û, vitamin Eƒ AST w ƒ w z ùkû, vitamin Eƒ

vitamin E Á» wp z 291 Table 4. The liver tissue total antioxidant of ICR mice that were treated by immobilization stress with or without vitamin E C 15.63±1.36 b, 13.26±0.84 b 2.783*, E 16.25±2.12 a 15.36±1.06 a 4.214* S+E 14.81±2.30 b 12.89±2.46 b 2.245 S 09.32±2.30 c 07.86±1.92 c 6.152** different (p<0.05) by Duncan s multiple range test. Values are meanq1 *: p<0.05, **: p<0.01 21,2 p w ù x y y w. TAS w y. 5 8 p ƒw z, y 18 z x TG, HDL-cholesterol š AST w.» y. TAS w d Table 4. 7 p S-7 9.32 mmol/l 15.63 mmol/l w 67% w, p ƒw vitamin E nw S+E-7 14.81 mmol/l p w 58% ƒw. 7 18 p S-7 9.32 mmol/l, S-18 7.86 mmol/l (t=6.15. w vitamin E n E-7 E-18 C-7 C-18. TAS 7 18 p TAS 2 w û ùkû. w Lovin š p w ROSƒ š ƒ s w s Á» y ƒ s q š sü SOD GSH-Px q. S-18 C-18, S+E-18, E-18 TAS w 50% w, 5 8 ƒw p w wy l DNA ùkù ƒ». p ƒw 24,25) vitamin E nw S+E-7, S+E-18 TAS ƒ z y vitamin Eƒ s q ww q. w s yƒ ƒ s q w, E-7 C-7 24) TAS vitamin Eƒ w y w». E-18 C- 18 TAS 8 vitamin E nw q» j z w ùkù q. SOD w y. TAS w SOD w. y w w z 26) SOD d Table 5. 7 p S-7 62.35 µ/ml C-7 w 35% wš, p ƒw vitamin E nw Table 5. The liver tissue superoxide dismutase of ICR mice that were treated by immobilization stress with or without vitamin E C 097.15±5.12 b, 90.01±3.91 a 04.837**, E 112.61±6.24 a 80.12±3.72 b 11.217*** S+E 080.12±6.13 c 71.21±5.32 c 06.341** S 062.35±8.21 d 41.20±4.16 d 04.016* *: p<0.05, **: p<0.01, ***: p<0.001 S+E-7 80.12 µ/ml p S-7 w 28% ƒw. 7 18 p S-7 62.35 µ/ml, S-18 41.20 µ/ml (t=4.016), S-18 C-18 SOD w 54%. SOD p w SODƒ Yun 27) w. pƒ sy w y ƒyg wyz SODy w e q, p vitamin E yw SOD ƒ vitamin Eƒ w sy w wy w s» yl w»w. 28) 7 18 p S-7 S-18 mw 20 pƒ TG, TAS, HDLcholesterol» SOD w e ùkû. š vitamin E nw E-7 C-7 SOD dw vitamin E SOD d ƒ, E-18 24) C- 18 û vitamin E n SOD ƒw ù mƒ v wš ƒ. MDA w y. MDA ü y p dw» w t wù y y. 29) TAS, SOD w MDA w dw. Table 6. 7 p S-7 8.98 nmol/l C-7 84% ƒwš, p ƒw vitamin E nw S+E-7 7.98 nmol/l p 11% w z. S+E-7 p S-7 w, S+E-18 p S-18 w û ùk ü. w C-18 C-7 20%. ù 20 ww 7 p MDAeƒ ùkù p MDA w ƒw Bedossa P 30) w. p x ƒw š vitamin E nw E-8 5.86 nmol/l w ùkû

292 ÁšÁ Table 6. The liver tissue malondialdehyde of ICR mice that were treated by immobilization stress with or without vitamin E C 0,4.86±0.91 d, 5.85±0.71 c -8.962***, E 5.86±0.74 c 5.97±0.69 c -0.215 S+E 7.98±0.69 b 8.13±0.91 b -0.816 S 8.98±0.71 a 9.10±0.83 a -0.342 ***: p<0.001 š, w x vitamin E nƒ g ù kù q. vitamin Eƒ p w, S+E-7 p S-7 w MDA û C-7 w vitamin E MDA. ù š mƒ vw üš ƒ. 18 p MDA eƒ C-18, E-18, S+E-18 ùkù, pƒ» MDA w ƒ w. pƒ z y transforming growth factor-β 1 (TGF-β 1 ) x ƒk, p w TGF-β 1 w me jš, α 1 (I) procollagen x ƒ, paracrine effect s(hepaticstellate cell) xƒƒ MDA e ƒk q. 31,3 p ƒw vitamin E nw S+E MDA w vitamin E w, y y vitamin Eƒ zƒ w. 33,34) vitamin Eƒ TGF-β 1 x g ROS w y MDA w wš q. w p w DNA vitamin Eƒ w35) MDA w z p MDA w w, p w s DNA y dw TG, HDL-cholesterol, AST, TAS, SOD Á» w q. 5 8 w 40 p ƒw x TG, HDL, AST TAS, SOD, MDA dw ù w e, 18 ¾ z x w š y w HDL-cholesterol ƒ 7%, SOD ƒ 33% y ùkûš MDA w 1.3%. p ù w pƒ». yw, 20» pƒ z p 40 60 ùkú w» pƒ s w w š. š p vitamin Eƒ z vitamin E x wš ƒ.» p vitamin E n ƒ p ù Á» w. ICR ù 20 w 5 ¾ w z, 8 p ƒwš ù» w 18 ¾ w p w ü y t x y w., p Á» x TG, AST w ƒjš, HDL-cholesterol w wj w,» vitamin E n p x TG AST w jš, HDL-cholesterol w ƒj z. p TAS, SOD w MDA w ƒ»g, p vitamin E n TAS, SOD w jš, MDA w wj w. w p» TG, HDLcholesterol, AST, TAS, SOD š MDA e w e ùkû. w mw w» p ù»¾ x w vitamin E yw z ùkü. Key words: p, k E, x, šx 1. Ghrousos, G. P. (2000) The role of stress and the hypothalamicpituitary-adrenal axis in the pathogenesis in the metabolic syndrome: Neuroendocrine and target tissue-related causes. Int. J. Obesity. 24, S50-S55. 2. Elliot, G. R. and Eisdorfer, C. (198 Stress and human health. New York, Springer, 2, 1-20. 3. Thomson, C. B. (1995) Apoptosis in the pathogenesis and treatment of disease. Science. 267, 1456-1462. 4. Gutteridge, J. N. C. (1995) Lipid peroxidation and antioxidants as biomarker of tissue damages. Clin. Chem. 41, 1819-1828. 5. Chow, C. K. (199 Vitamin E and oxidative stress. Free Radical Bio. Med. 11, 215-232. 6. Tiidus, P. M. and Houston, M. E. (1995) Vitamin E status and response to exercise training. Sports Med. 20, 12-23. 7. Kagan, V. E., Spirichev, V. B. and Serbinova, E. A. (1994) The significance of vitamin E and free radicals in physical exercise.

vitamin E Á» wp z 293 In Nutrition in exercise and sport, Wolinsky, I. (2nd ed.) pp. 185-213, CRC Press, Boca Raton. 8. Avellini, L., Chiaradia, E. and Gaiti, A. (1999) Effect of exercise training, selenium and vitamin E on some free radical sacvengers in horse. Comp. Biochem. Phys. B. 123, 147-154. 9. Livne, E., Laufer, D. and Blumenfeld, I. (1997) Comparison of in vitro response to growth hormone by chondrocytes from mandibular condyle cartilage of young and old mice. Calcified. Tissue. Int. 7, 61-62. 10. Park, S. Y. (200 Study on the oxidative damage induced by stress and its recovery in mice. Department of Microbial Engineering. Dissertation of Ph. D. Konkuk Universtiy. Seoul. 11. Kissebah, A. H. (1974) Stress hormines and lipid metabolism. Proc. R. Soc. Med. 67, 665-667. 12. Howard, N. H., Dieter, M. K., Poetro, A., Gabriele, B. B., Giuseppe, G., Juliana, H., Hazel, P. and Alex, S. (1994) Biochemical and cytotoxic characterstics of an in vivo circulating oxidized LDL. J. Lipid Res. 35, 669-677. 13. Collett, C., Pares-Herbute, N., Monnier, L. H. and Cartry, E. (1988) Platelet function in type I diabetes: Effects of supplementation with large doses of vitamin E. Am. J. Clin. Nutr. 47, 256-261. 14. Gittleman, B., Shatin, L., Bierenbaum, M. L., Fleisohman, A. I. and Hqyton, T. (1968) Effects of quantified stressful stimuli on blood lipids in man. J. Nutr. Ment. Dis. 147, 196-201. 15. Lehtonen, A. J. (1978) The effect of vigorous physical activity at work on serum lipids with a special reference to serum highdensity lipoprotein cholesterol. Acta. Physiol. Scand. 104, 117-121. 16. Kruit, J. K., Groen, A. K., van Berkel, T. J. and Kuipers, F. (2006) Emerging roles of the intestine in control of cholesterol metabolism. World. J. Gastroenterol. 12, 6429-6439. 17. Lim, S. S., Kim, M. H. and Lee, J. H. (1997) Effect of Artemisia princeps var orientalis and Circium japonicum var ussuriense on liver function body lipid and bile acid of hyperlipidemic rat. Korean J. Nutr. Soc.. 30, 797-802. 18. Woo, Y. S. (199 Studies on the effects of slaughter stress on the blood picture and serum chemical values of pigs. Dissertation of Ph. D. Konkuk Universtiy. Seoul. 19. Hernry, J. B. (2007) Measurement of lipids and evaluation of lipid disorders. In Clinical diagnosis and management by Laboratory Methods, Levy, M. D. pp. 171-198, Elsevier Science Health Science div, Churchill. 20. Rao, M. S., Pareddy, K., Abecassis, M. and Hashimoto, T. (200 Regeneration of liver with marked fatty change following partial hepatectomy in rats. Digest. Dis. Sci. 46, 1821-1826. 21. Cherubini, A., Zuliani, G. and Costantini, F. (200 High vitamin E plasma levels and low low-density lipoprotein oxidation are associated with the absence of atheros-clerosis in octogenarians. J. Am. Geriatr. Soc. 49, 651-654. 22. Gale, C. R., Ashurst, H. E., Powers, H. J. and Martyn, C. N. (200 Antioxidant vitamin status and carotid atherosclerosis in the elderly. Am. J. Clin. Nutr. 74, 402-408. 23 Lovin, R., Cottle, W., Pyke, I., Davanagh, M. and Belcastro, A. N. (1987) Are induces of free radical damage related to exercise intensity. Eur. J. Appl. Physiol. 6, 313-316. 24. Lawler, J. M., Powers, S. K., Dijk, H. V., Visser, T., Kordus, M. J. and Ji, L. L. (1994) Metabolic and antioxidant enzyme activities in the diaphragm: effects of acute exercise. Res. Physiol. Neurobiol. 96, 139-149. 25. Ji, L. L. (199 Blood glutathione status during exercise; Effect of carbohydrate supplementation. J. Appl. Physiol. 74, 788-792. 26. Halliwell, B. and Gutteridge, J. M. C. (1989) In Free radicals in biology and medicine, (2nd ed.) Clarendon Press, Oxford. 27. Yun Mei, K. R., Gawai., Zhongzhen, Nie., Vickmar. and Robert, H. Helfert. (1999) Age related reductions in the activities of antioxidant enzymes in the rat inferior colliculus. Hearing. Res. 135, 169-180. 28. Scarpa, M., Rigo, A., Maiorino, M., Ursini, F. and Gregolin, C. (1984) Formation of α-tocopherol radical and recycling of α- tocopherol by ascorbate during peroxidation of phophatidylcholin liposomes. An electron paramagnetic resorbance study. Biochem. Biophys. Res. Comm. 801, 215-219. 29. Okimoto, K., Hamazaki, K., Lwagaki, H., Orita, K. and Mori, A. (1995) Effect of picibanil on the scavenging effect of free radicals produced during liver regeneration in the rat. Acta. Med. Okayama. 49, 75-79. 30. Bedossa, P., Houglum, K., Trautwein, C., Holstege, A. and Chojkier, M. (1994) Stimulation of collagen α I(I) gene espression is associated with lipid peroxidation in hepatocellular injury: a Link tot issue fibrosis. Hepatology. 19, 1262-1271. 31. Nakatsukasa, H., Evarts, R. P., Hsia, C. C. and Thorgeirsson, S. S. (1990) Transforming growth factor-β 1 and type I procollagen transcripts during regeneration and early fibrosis of rat liver. Lab Invest. 63, 171-180. 32. Armendariz-Borunda, J., Seyer, J. M., Kang, A. H. and Raghow, R. (1990) Regulation of TGF-β gene espression in rat liver intoxicated with carbon tetrachloride. FASEB J. 4, 215-221. 33. Brenner, D. A., Veloz, L., Jaenisch, R. and Alcorn, J. M. (199 Stimulation of the collagen α I (I) endogenous gene and transgene in carbon tetrachloride-induced hepatic fibrosis. Hepatology. 17, 287-292. 34. Castilla, A., Prieto, J. and Fausto, N. (199 Transforming growth factor β1 and α in chronic liver disease. N. Eng. J. Med. 324, 933-940. 35. Traber, M. G. (200 Does vitamin E decrease heart attack risk summary and implications with respect to dietary recommendations. J. Nutr. 131, 395-397.