Journal of the East Asian Society of Dietary Life (동아시아식생활학회지)

The East Asian Society of Dietary Life (동아시아식생활학회)
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Hepatoprotective Effects of Fermented Smilax china Leaf Extract on Carbon Tetrachloride-induced Liver Injury in Mice

발효 청미래덩굴잎 추출물이 사염화탄소에 의한 마우스의 간 손상 보호 효과

  • Kim, Mee-Jung (Division of Hotel Food & Bakery, Shinsung University)
  • 김미정 (신성대학교 호텔조리제빵계열)
  • Received : 2014.02.07
  • Accepted : 2014.04.16
  • Published : 2014.04.30
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Abstract

This study was conducted to investigate the hepatoprotective effects of fermented Smilax china leaf ethylacetate extracts by Aspergillus oryzae on carbon tetrachloride-induced liver injury in mice. Experimental mice were divided into four groups (five mice/group) (NC; normal control group, CB; basic diet supplemented before $CCl_4$ treatment group, NS ; basic diet mixed with 0.5% Smilax china leaf ethyl acetate extract supplemented before $CCl_4$ treatment group, FS; basic diet mixed with 0.5% fermented Smilax china leaf ethyl acetate extract supplemented before $CCl_4$ treatment group) fed for 4 weeks each. In the $CCl_4$-treated groups (CB, NS and FS) compared with the NC group, liver weights, activities of alanine aminotransferase, aspartate aminotransferase, xanthine oxidase and aldehyde oxidase, contents of triglycerides, total cholesterol and LDL-cholesterol in serum, and hepatic lipid peroxide levels increased, whereas body weight gain and contents of glutathione and HDL-cholesterol decreased. Furthermore, in the FS groups compared with the NS and CB groups, increased or decreased indicators by $CCl_4$ treatment significantly decreased or increased, respectively. This study suggests that fermented Smilax china L. leaf extracts may regulate xanthine oxidase and aldehyde oxidase inhibitory activities and hepatoprotective effects due to flavonoid aglycone derived from its glycoside in leaves of Smilax china by fermentation of A. oryzae.

발효 청미래덩굴잎의 해독효과를 조사하기 위하여 정상식이군(NC), 기본식이를 급여한 후 $CCl_4$ 처리군(CB), 비발효 청미래덩굴잎 ethylacetate 추출 분획물을 기본 식이에 0.5% 혼합한 식이로 4주간 사육하였다. $CCl_4$를 처리한 군(NS), Aspergillus oryzae로 발효시킨 청미래덩굴잎 ethylacetate 추출 분획물을 기본 식이에 0.5% 혼합한 식이로 4주간 사육한 후, $CCl_4$를 처리한 군(FS)의 4개군(5마리/군)으로 구분하여 체중, 식이섭취량, 식이효율, ALT 및 AST 활성, GSH 및 LPO 함량, 혈청 지질 함량, XO 및 AO 활성도를 조사하였다. $CCl_4$를 처리한 군(CB, NS, FS)의 체중증가량은 NC군에 비하여 현저하게 감소하였으나, NS군과 FS군에서는 그 감소율이 낮았으며, FS군에서 더욱 낮았다. 이와 반대로 간의 중량은 $CCl_4$ 처리군들에서 높았으나, FS군에 NS군에 비하여 낮았다. ALT 및 AST 활성은 CB군이 NC군에 비하여 32.34~3.80배가 높았으며, NS군 및 FS군은 CB군에 비하여 각각 16.34~37.35% 및 34.97~45.66%가 낮았다. 혈청 중성 지질의 함량은 NC군에 비하여 CB군이 81.71%가 높았으며, NS군과 FS군은 CB군에 비하여 각각 15.61% 및 28.22%가 낮았다. Total cholesterol 함량은 CB군이 NC군에 비하여 54.07%가 높았으나, NS군 및 FS군은 CB군에 비하여 각각 10.59% 및 17.99%가 낮았으며, LDL-cholesterol의 함량도 유사한 경향이었다. 반면에, HDL-cholesterol의 함량은 CB군이 NC군에 비하여 55.08%가 낮았으나, NS군 및 FS군은 CB군에 비하여 각각 26.96% 및 72.12%가 높았다. GSH의 함량(${\mu}mole/g$ of tissue)은 CB군(4.42)이 NC군(5.09)에 비하여 13.16%가 낮았으나, NS군과 FS군은 각각 4.84 및 4.93으로 NC군과의 유의적인 차이를 보이지 않았으며, NS군과 FS군 간에도 뚜렷한 차이를 보이지 않았다. 그러나 LPO의 함량(MDA nmole/g of tissue)은 CB군이 NC군에 비하여 36.71%가 높았으나, 특히 FS군에서는 NC군 수준으로 회복되었다. XO(xanthine oxidase) 활성(% against control)은 NC군이 98.45, CB군 128.97, NS군 119.70, FS군 90.65로 NS군은 CB군과, FS군은 NS군과의 유의적인 차이를 보이지 않았다. AO(aldehyde oxidase)의 활성(% against control)은 CB군이 NC군에 비하여 22.48%가 높았으나, NS군 및 FS군은 NC군에 비하여 각각 14.88% 및 7.56%가 높았으며, CB군에 비하여는 각각 8.94% 및 16.15%가 감소하였다. 이상의 결과, Aspergillus oryzae로 발효시킨 청미래덩굴잎 ethylacetate 추출물은 비발효 경우에 비하여 $CCl_4$를 처리한 mouse에서 간 조직의 손상을 회복시키는 효과가 높으며, 이는 FS군에서 XO 및 AO의 저해활성이 높은 것으로 미루어, 청미래덩굴잎에 존재하는 flavonoid 배당체가 발효에 의하여 분해되어 aglycone화함으로써 나타난 결과라 사료된다.

Keywords

References

  1. Ali S, Pawa S, Naime M, Prasad R, Ahmad T, Farooqui H, Zafar H (2008) Role of mammalian cytosolic molybdenum Fe-S flavin hydroxylases in hepatic injury. Life Sci 82: 780-788. https://doi.org/10.1016/j.lfs.2008.01.011
  2. Amdur MO, Doull J, Klaassen CD (1980) Toxicology. 2th ed. Pergamon Press, New York, USA, p 210-231.
  3. Arthus CG (1991) Textbook of Medical Physiology. 8th ed. Sanders PA, USA, p 754-764.
  4. Carr MC (2003) The emergence of the metabolic syndrome with menopause. J Clin Endocrinol Metab 88: 2404-2411. https://doi.org/10.1210/jc.2003-030242
  5. Cha BC, Lee EH (2007) Antioxidant activities of flavonoids from the leaves of Smilax china Linne. Kor J Pharmacogn 38(1): 31-36.
  6. Cho SY, Jang JY, Kim MJ (2001) Effects of Pueraia flos and radix water extracts on levels of several serum biomarkes in theanol-treated rats. J Korean Soc Food Sci Nutr 30: 92-96.
  7. Dambrova M, Uhle'n S, Welch CJ, Wikberg JES (1998) Identification of an N-hydroxyguanidine reducing activity of xanthine oxidase. Eur J Biochem 257: 178-184. https://doi.org/10.1046/j.1432-1327.1998.2570178.x
  8. Dong H, Hailing RL, Thummel KE, Rettie AE, Nelson SD (2000) Involvement of human cytochrome P450 2D6 in the bioactivation of acetaminophen. Drug Metab Dispos 28, 1397-1400.
  9. Ellman GL (1959) Tissue sulfhydryl group. Arch Biochem Biophys 82: 70-77. https://doi.org/10.1016/0003-9861(59)90090-6
  10. Fridewald WT, Levy RI, Fredrickson DS (1972) Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem 18: 499-502.
  11. Garattini E, Fratelli M, Terao M (2009) The mammalian aldehyde oxidase gene family. Hum Genomics 4: 119-130.
  12. Guo J, Qian F, Li J, Xu Q, Chen T (2007) Identification of a new metabolite of astilbin, 3-O-methylastilbin, and its immunosuppressive activity against contact dermatitis. Clin Chem 53(3): 465-471. https://doi.org/10.1373/clinchem.2006.077297
  13. Halder B, Pramanick S, Mukhopadhyay S, Giri AK (2006) Anticlastogenic effects of black tea polyphenols theaflavins and thearubigins in human lymphocytes in vitro. Toxicol In Vitro 20: 608-613. https://doi.org/10.1016/j.tiv.2005.10.010
  14. Hamzeh-Mivehroud M, Rahmani S, Rashidi MR, Hosseinpour Feizi MA, Dastmalchi S (2013) Structure-based investigation of rat aldehyde oxidase inhibition by flavonoids. Xenobiotica 43(8): 661-670. https://doi.org/10.3109/00498254.2012.755228
  15. Hue FS, Krook L, Pond WG, Duncan JR (1975) Interactions of dietary calcium with toxic levels of lead and zinc in pigs. J Nutr 105: 112-118.
  16. Imai K, Nakachi K (1995) Cross sectional study of effects of drinking green tea on cardiovascular and liver diseases. Br Med J 310: 693-696. https://doi.org/10.1136/bmj.310.6981.693
  17. Itoh K (2009) Individual and strain differences of aldehyde oxidase in the rat. Yakugaku Zasshi 129: 1487-1493. https://doi.org/10.1248/yakushi.129.1487
  18. Janbaz KH, Saeed SA, Gilani AH (2002) Protective effect of rutin on paracetamol- and CCl4-induced hepatotoxicity in rodents. Fitoterapia 73: 557-563. https://doi.org/10.1016/S0367-326X(02)00217-4
  19. Jeong TC, Kim HJ, Park JI (1997) Protective effects of red ginseng saponins against carbon tetrachloride-induced hepatotoxicity inspargue Dawley rats. Planta Med 63: 136-140. https://doi.org/10.1055/s-2006-957630
  20. Jung JH (2011) Risk factors of environmental diseases in an industrial complex area. Korean J Ind Heal 5: 69-77.
  21. Karmen A (1955) A note on the spectrophotometric assay of glutamic oxaloacetic transaminase in human blood serum. J Clin Invest 34: 131-133.
  22. Krenitsky TA (1978) Aldehyde oxidase and xanthine oxidasefunctional and evolutionary relationships. Biochem Pharmacol 27: 2763-2764. https://doi.org/10.1016/0006-2952(78)90186-7
  23. La Casa C, Villegas I, Alarcón de la Lastra C, Motilva V, Martín Calero MJ (2000) Evidence for protective and antioxidant properties of rutin, a natural flavone, against ethanol induced gastric lesions. J Ethnopharmacol 71: 45-53. https://doi.org/10.1016/S0378-8741(99)00174-9
  24. Lee JS, Kim HS, Lee YJ, Yong CS, Choi HG, Han GD (2008) Hepatoprotective effect of Grifola frondosa water extract on caarbon tetrachloride-induced liver injury in rats. Food Sci Biotechnol 17(1): 203-207.
  25. Lee SI, Lee YK, Kim SD, Kang YH, Suh JW (2012) Antioxidative activity of Smilax china L. leaf teas fermented by different strains. Korean J Food Nutr 25(4): 807-819. https://doi.org/10.9799/ksfan.2012.25.4.807
  26. Lee YK, Lee Sl, Kim JS, Yang SH, Lee IA, Kim SD, Suh JW (2012) Antioxidant activity of green tea fermented with Monascus pilosus. J Appl Biol Chem 55(1): 19-25. https://doi.org/10.3839/jabc.2011.054
  27. Lowry OH, Rosebrough NJ, Farr AL, Randall RL (1951) Protein measurement by folin phenol reagent. J Biol Chem 193: 265-275.
  28. McGregor D, Lang M (1996) Carbon tetrachloride: Genetic effects and other modes of action. Mutation Res 366: 181-195
  29. Nishino T, Tamura I (1991) The mechanisim of conversion of xanthine dehydrogenase to oxidase and the role of the enzyme in the resperfusion injury. Adv Exp Med Biol 309: 327-337.
  30. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95: 248-254.
  31. Parks DA, Granger DN (1986) Xanthine oxidase: Biochemistry, distribution and physiology. Acta Physiol Scand Suppl 548: 87-99.
  32. Park JW, Lee HH, Oh MS, Kim JP, Jang TK, You YN, Ha DR, Kim ES, Seo KW (2013) A survey on pesticide residues of commercial flowering teas. J Korean Pesticide Science 17(1): 1-15. https://doi.org/10.7585/kjps.2013.17.1.1
  33. Park SJ, Kang MH (2003) The effect of dietary nuddle with glucomannan on the weight loss in high fat diet-induced obese rats. J Korean Soc Sci Nutr 32: 893-898. https://doi.org/10.3746/jkfn.2003.32.6.893
  34. Park SN (2010) A study on entry strategies of Korean green tea into the world market. Wonkwang University Graduate School of Oriental Studies.
  35. Pierce RA, Glaug MR, Greco RS, Mackenzie JW, Boyd CD, Deak SB (1987) Increased procollagen mRNA levels in $CCl_4$-induced liver fibrosis in rats. J Biol Chem 262: 1652- 1658.
  36. Rajagopalan KV, Fridovich I, Handler P (1962) Hepatic aldehyde oxidase. I. Purification and properties. J Biol Chem 237: 922-928.
  37. Recknagel RO, Glende Jr EA, Dolak JA, Waller RL (1989) Mechanisms of carbon tetrachloride toxicity. Pharmac Ther 43:139-154. https://doi.org/10.1016/0163-7258(89)90050-8
  38. Reitman S, Frankel S (1957) A colorimetric method for the determination of serum glutamic oxaloacetic pyruvic transaminase. AM J Clin Pathol 28: 58-63.
  39. Rott KT, Agudelo CA (2003) Gout. J Am Med Assoc 289: 2857-2860. https://doi.org/10.1001/jama.289.21.2857
  40. Roy R, McCord JM (1982) Ischemia-induced conversion of xanthine dehydrogenase to xanthine oxidase. Fed Proc 41: 767-773.
  41. Shaw S, Jayatilleke E (1990) The role of aldehyde oxidase in ethanol-induced hepatic lipid peroxidation in the rat. Biochem J 268: 579-583.
  42. Shao B, Guo HZ, Gui YJ, Ye M, Han J, Guo DA (2007) Steroidal saponins from Smilax china L. and their anti-inflammatory activities. Phytochem 68: 623-630. https://doi.org/10.1016/j.phytochem.2006.10.026
  43. Stirpe F, Della Corte E (1969) The regulation of rat liver xanthine oxidase. Conversion in vitro of the enzyme activity from dehydrogenase (type D) to oxidase (type O). J Biol Chem 244: 3855-3863.
  44. Sung IS, Kim MJ, Cho SY (1997) Effect of Quercus acutissima Carruthers extracts on the lipid metabolism. J Korean Soc Food Sci Nutr 26: 327-333.
  45. Stoddart AM, Levine WG (1992) Azoreductase activity by purified rabbit liver aldehyde oxidase. Biochem Pharmacol 43: 2227-2235. https://doi.org/10.1016/0006-2952(92)90182-I
  46. Weber LWD, Boll M, Stampfl A (2003) Hepatotoxicity and mechanism of action of haloalkanes: Carbon tetrachloride as a toxicological model. Crit Rev Toxicol 33: 105-136. https://doi.org/10.1080/713611034
  47. Weigert J, Neumeier M, Bauer S, Mages W, Schnitzbauer AA, Obed A, Gröoschl B, Hartmann A, Schäaffler A, Aslanidis C, Schöolmerich J, Buechler C (2008) Small-interference RNA-mediated knock-down of aldehyde oxidase 1 in 3T3-L1 cells impairs adipogenesis and adiponectin release. FEBS Lett 582: 2965-2972. https://doi.org/10.1016/j.febslet.2008.07.034
  48. Williams AT, Burk RF (1990) Carbon tetrachloride hepatotoxicity: An example of free radical-mediated injury. Seminars Liver Dis 10: 279-284. https://doi.org/10.1055/s-2008-1040483
  49. Yuan LP, Chen FH, Ling L, Dou PF, Bo H, Zhong MM, Xia LJ (2008) Protective effects of total flavonoids of Bidens pilosa L. (TFB) on animal liver injury and liver fibrosis. J Ethnopharmacol 116(3): 539-546. https://doi.org/10.1016/j.jep.2008.01.010