키토산올리고당이 카드뮴을 투여한 생쥐의 간독성에 미치는 효과

Effects of Chitosanoligosaccharide on the Hepatotoxicity in Cadmium-treated Mice

  • 윤중식 (조선대학교 자연과학대학 생물과학부) ;
  • 노영복 (조선대학교 자연과학대학 생물과학부)
  • Yoon, Jung-Sik (Department of Biological Sciences, College of Natural Sciences, Chosun University) ;
  • Roh, Young-Bok (Department of Biological Sciences, College of Natural Sciences, Chosun University)
  • 발행 : 2003.03.01

초록

본 연구는 카드뮴으로 유발된 생쥐 간중독에 대한 키토산올리고당의 효과를 알아보기 위하여 시도되었다. Mouse를 대상으로 카드뮴 (5.0 mg/kg; i.p.) 단독투여군(group Cd), 카드뮴과 키토산올리고당 (0.5% solution) 동시투여군 (group Cd+Chi) 그리고 키토산올리고당 1주일 전처치군 (group Chi7+Cd)으로 구분한 후 간손상 억제효과를 알아보기 위해 혈청중 효소활성도와 glutathione peroxidase (GSH-Px)의 활성도 그리고 glutathione reductase (GR)의 활성도를 비교측정하였다. 또한 간조직의 형태학적 손상을 확인하기 위해 조직학적 관찰을 실시하였다. 혈청 효소활성결과, AST, ALT 그리고 LDH의 활성도는 키토산올리고당을 처치한 Cd+Chi군과 Chi7+Cd군이 Cd군보다 감소되었다. 그러나 TP와 Alb의 활성도는 키토산올리고당을 처치한 Cd+Chi군과 Chi7+Cd군이 Cd군보다 증가되었다. GSH-Px활성은 Cd군에 비해 Cd+Chi군과 Chi7+Cd군이 현저하게 증가되었다. GR활성은 Cd군에 비해 Cd+Chi군과 Chi7+Cd군이 현저하게 감소하였다. 광학현미경적 관찰 결과, Cd군은 카드뮴 독성에 의한 간세포의 괴사가 관찰되었다. Cd+Chi군과 Chi7+Cd군은 정상군과 유사한 소견을 보였다. 이상과 같은 결과로, 키토산올리고당이 생쥐 간조직에 미치는 카드뮴의 독성을 감소시킴을 알 수 있었다.

This research was conducted to determine the effects of chitosanoligosaccharide on liver poisoning induced by cadmium (Cd). Three groups of mice were used in this research. The first group was only injected with cadmium (5.0 mg/kg; i.p.) (group Cd) and the second one with cadmium and chitosanoligosaccharide (0.5% solution) at the same time (group Cd+Chi). The third one which had already been injeted with chitosanoligosaccharide (0.5% Solution) aweek before (group Ch7+Cd) was used. In order to investigate the inhibitory action of chitosanoligosaccharide on liver damage, enzyme activity in serum, glutathione peroxidase (GSHPx) activity and glutathione reductase (GR) activity were relatively measured. In addition, histological observations were made to determine the morphologic injury of liver tissues. As the result of enzyme activity in serum, the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) in chitosanoligosaccharide-injected groups Cd+Chi and Chi7+Cd was lower than in group Cd. GSH-Px activity was sharply increased in groups Cd+Chi and Chi7+Cd compared to group Cd. GR activity was conspicuously decreased in groups Cd+Chi and Chi7+Cd compared to group Cd. As the result of light microscopic observation, liver cell necrosis caused by cadmium poisoning was obseved in liver cells. The finding of group Cd+Chi and Chi7+Cd was similar total on of normal groups. As the result of electron microscopic observation, mitochondria in group Cd showed a severe swelling phenomenon, RER fragment and ribosome dropout. However, in groups Cd+Chi and Chi7+Cd, mitochondria wiht high electron density were distributed and RER forming a typical lamellae with ribosome was observed. From these results, cadmium toxicity on rat liver tissues could be lessened by chitosanoligosaccharide.

키워드

참고문헌

  1. Amako K, Shimodori S, Imoto T, Miake S, UmedaA: Effects of chitin and its soluble derivatives on survival of Vibrio cholerae O1 at low temperature, Appl Environ Microbiol 53(3) : 603 605, 1987
  2. Anderson HR, Anderson O: Effect of cadmium on hepatic lipid peroxidation in mice, Pharmacol Toxicol 63 173 177, 1988 https://doi.org/10.1111/j.1600-0773.1988.tb00934.x
  3. Basavaraju SR, Jones TD: Atherosclerotic risks from chemicals : Part I, Toxicological observations and mechanisms of atheroscerosis, Arch Environ Contam Toxicol 35 152 164, 1998 https://doi.org/10.1007/s002449900363
  4. Bough WA, Landes DR: Recovery and nutritional evaluation of proteinaceous solids separated from whey by coagulation with chitosan, J Dairy Sci 59(11) ',1874 1880, 1976 https://doi.org/10.3168/jds.S0022-0302(76)84456-6
  5. Brzoska MM, Moniuszko Jakoniuk J, Jurczuk M, Galazyn Sidorczuk M, Rogalska J: The effect of zinc supply on cadmiwn induced changes in the tibia ohats, Food Chern Toxicol 39 : 729 737, 2001 https://doi.org/10.1016/S0278-6915(01)00007-2
  6. Carlberg I, Mannervik B: Glutathione reductase, Methods Enzymol 113 :484 490, 1985 https://doi.org/10.1016/S0076-6879(85)13062-4
  7. Cartana J, Romeu A, Arola L: Effects of copper, cadmium and nickel on liver and kidney glutathione redox cycle of rats, Comp Biochem Physiol 101(2) : 209 13, 1992 https://doi.org/10.1016/0300-9629(92)90524-T
  8. Chan HM, Cherian MG: Protective roles of metallothionein and glutathione in hepatotoxicity of cadmium, Toxicology 72(3) : 281 290, 1992 https://doi.org/10.1016/0300-483X(92)90179-I
  9. Chen JP, Chen JY: Preparation and characterization of immobilized phospholipase Az on chitosan beads for lowering serwn cholesterol concentration, J Mole Cataly B: Enzymatic 5: 483 490, 1998 https://doi.org/10.1016/S1381-1177(98)00077-0
  10. Cherian MG, Nordberg M: Cellular adaptation in metal toxi- cology and metallothionein, Toxicology 28 : 1 15, 1983 https://doi.org/10.1016/0300-483X(83)90101-4
  11. Cherroret, G.;Dessor, O.;Lehr, P.R.: In vit!'o effects of alwninum chloride on choline acetyltransferase activity of the rat brain during postnatal growth, Bull Environ Contam Toxicol 52: 487 490, 1994
  12. Davies, P.;Maloney, A.F.J.: Selective loss of cholibergic neurones in alzheimer's disease, Lancet 2: 1403 1409, 1976
  13. Deuchi K, Kanauchi O, Shizukuishi M, Kobayashi E: Continuous and massive intake of chitosan affects mineral and fat soluble vitamin status in rats fed on a high fat diet, Biosci Biotechnol Biochem 59(7) : 1211 1216, 1995 https://doi.org/10.1271/bbb.59.1211
  14. Dudley RE, Svoboda DJ, Klaassen CD: Acute exposure to cadmium causes severe liver injury in rats, Toxicol Appl Pharmacol 65(2) : 302 313, 1982 https://doi.org/10.1016/0041-008X(82)90013-8
  15. Dudley RE, Svoboda DJ, Klaassen CD: Time course of Cd induced ultrastructural change in rat liver. Toxicol Appl Pharmacol 76',150 160, 1984 https://doi.org/10.1016/0041-008X(84)90038-3
  16. Dureja H, Tiwary AIL, Gupta S: Simulation of skin penneability in chitosan membranes, Int J Pharm 213(12) : 1938, 2001
  17. Engastrom B, Nordberg GF: Effects of detergent fomular chelating agent on the metabolism and toxicity of cadmium in mice, Acta Pharmacol Toxicol 43 : 387 397, 1978
  18. Garrett Sa, Phillips V, Somji S, Sens MA, Dutta R, Park SM, Kim DY, Sens DA: Transient induction of metallothionein isoform 3(MT 3), c los, c jun and c myc in human proximal tubule cells exposed to cadmium, Toxicol Lett 126: 69 80, 2002 https://doi.org/10.1016/S0378-4274(01)00448-9
  19. Goering PL, Klaassen CD: Altered subcellular distribution of cadmium following cadmium protreatment: possible mechanism of tolerance to cadmiwn induced lethality, Toxicol Appl Pharmacol 70: 195 203, 1983 https://doi.org/10.1016/0041-008X(83)90095-9
  20. Goering PL, Klaassen CD: Tolerance to cadmiwn induced hepatotoxicity following cadmium pretreahnent, Toxicol Appl Pharmacol 74(3) : 308 313, 1984 https://doi.org/10.1016/0041-008X(84)90283-7
  21. Goyer RA, Tsuchiya R, Leonard DL, Kalwo H: Aminoaciduria in japanese workers in the lead and cadmium industries, Am J Clin Pathol57 : 635 642, 1972 https://doi.org/10.1093/ajcp/57.5.635
  22. Guilhermino L, Soares AM, Carvalho AP, Lopes MC: Effects of cadmium and parathion exposure on hematology and bloodbiochemisty of adult male rats, Bull Environ Contm Toxicol 60(1) : 52 59, 1998 https://doi.org/10.1007/s001289900590
  23. Hoffinann EO, CookJA, di Luzio NR, Coover JA: The effects of acute cadmium administration in the liver and kidney of the rat, Light and electron microscopic studies, Lab Invest 32(5) : 655 664, 1975
  24. : Dexamethasone prevents acute cadmium induced hepatic injury but exacerbates kidney dysfunction in rabbits, ToxicolAppl PharmacoI 174(3) : 225 234, 2001
  25. Janes K.A, Calvo P, Alonso MJ: Polysaccharide colloidal particles as delivery systems for macromolecules, Adv Drug Deliv Rev 47(1) : 83 97, 2001 https://doi.org/10.1016/S0169-409X(00)00123-X
  26. Jeon YJ, Kim GH, Park PJ, Kim SK: Calcium absorption accelerating effect of chitosan oligosaccharides prepared by ultrafiltration membrane enzymatic reactor. J Kor Fish Soc 32(3) : 247 251, 1999
  27. Kannakar R, Chatterjee M: Cadmium induced time depen- dent oxidative stress in liver of mice: a correlation with kidney. Environ Toxicol Phannaco l6 : 201 207, 1998
  28. Kim HK, Kim KY, Born HS, Choi KH, Kim JY: Effect of oral injection of chitosan and alginate on the removal of orally injected radiostrontium (85Sr) in mice. J Kor Nucl Med 27: 130 134, 1993
  29. Kim YH, Bwn HS, Kim KY, Kim HL, Kim JY: Inhibitory effect of chitosan on the milk transfer of radiostrontium from contaminated mice to their suelders. J Chitin Chitosan 4(1): 15 18, 1999
  30. Kowel NE, Johnson DE, KaBmer OF, Pahren HR: Normal levels of cadmium in diet, urine, blood and tissues of inhabitants of the United States, J Toxicol Environ Health 5 :995 998, 1979 https://doi.org/10.1080/15287397909529809
  31. Kudo N, Yamashina S, Waku K: Protection against cadmium toxicity by zinc: Decrease in the Cd high molecular weight protein fraction in rat liver and kidney on Zn pretreatment. Toxicology 40(3) : 267 277, 1986 https://doi.org/10.1016/0300-483X(86)90059-4
  32. Lafuente A, Marquez N, Perez Lorenzo M, pazo D, Esquifino AI: Pubertal and postpubertal cadmium exposure differentially affects the hypothalamic pituitary testicular axis function in the rat, Food Chern Toxicol 38 : 913 923, 2000 https://doi.org/10.1016/S0278-6915(00)00077-6
  33. Landes DR, Bough WA: Effects of chitosan a coagulating agent for food processing wastes in the diets of rats on growth and liver and blood composition, Bull Environ Contam Toxicol 15(5) : 555 63, 1976 https://doi.org/10.1007/BF01685704
  34. Lawrence RA, Burk RF: Glutathione peroxidase activity in selenium deficient rat liver. Biochem Biophys Res Commun 71(4) : 952 958, 1976 https://doi.org/10.1016/0006-291X(76)90747-6
  35. Leber AP, Miya TS: A mechanism for cadmiwn and zinc induced tolerance to cadmium toxicity: Involvement of metallothionein, Toxicol Appl Phannacol 37: 403 414, 1976 https://doi.org/10.1016/0041-008X(76)90202-7
  36. Lee JS, White KL: A review of the health effects of cadmium, Am J Ind Med 1(34) : 307 317, 1980 https://doi.org/10.1002/ajim.4700010308
  37. Liu J, Liu Y, Sultan S, Habeebu SS, Klaassen CD: Metallo-thionein null mice are highly susceptible to the hemato-toxic and immunotoxic effects of chronic CdCI2 exposure, Toxicol Appl Phannacol 159: 98 108, 1999 https://doi.org/10.1006/taap.1999.8718
  38. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the folin phenol reagent. J Biol Chern 193: 265 275, 1951
  39. Medesani DA, Lopez Greco LS, Rodriguez EM: Effects of cadmium and copper on honnonal regulation of glycemia by the eyestalks in the Crab Chasmagnathus granulata, Bull Environ Contam Toxicol 66 : 71 76, 2001 https://doi.org/10.1007/s001280000207
  40. Morita S: Defense mechanisms against cadmium toxicity, Jpn J Phannacol 35(2) : 129 141, 1984
  41. Mutanen, ML., Mykkanen, HM.: Effect of dietary fat on plasma glutathione peroxidase levels and intestinal absorption of 75Se labeled sodium selenite in chicks, J Nutr 114(5) 829 834, 1984 https://doi.org/10.1093/jn/114.5.829
  42. Muzzarelli RAA, Ferrero A, Pizzoli M: Light scattering, Xray diffraction, elemental analysis and infrared spectropho-tometry characterization of chitosan, a chelating polymer, Talanta 19(10) : 1222 1226, 1972 https://doi.org/10.1016/0039-9140(72)80068-7
  43. Nakajima M, Atsumi K, Kifooe K, Miura K, Kanamaru H: Chitin is an effective material for sutures, JpnJ Surg 16(6) : 418 424, 1986 https://doi.org/10.1007/BF02470609
  44. Nishimura K, Ishihara C, Ukei S, Tokura S, Azuma I: Stimulation of cytokine production in mice using deacety-lated chitin, Vaccine 4(3) : 151 156, 1986 https://doi.org/10.1016/0264-410X(86)90002-2
  45. Nishimura Y, Kakuta I, Talceda H, InabaJ, Imai K, Watari K, Matsusalca N: Effect of natural chelating agents on the intestinal absorption of radiostrontium in rats, Radiat Protect Dosim 53 : 331 334, 1994 https://doi.org/10.1093/rpd/53.1-4.331
  46. Nordberg M: Studies on metallothionein and cadmium, Environ Res 15: 381 404, 1978 https://doi.org/10.1016/0013-9351(78)90120-2
  47. Novelli EL, Vieira EP, Rodrigues NL, Ribas BO: Risk assessment of cadmium toxicity on hepatic and renal tissues of rats, Environ Res 79(2) : 102 105, 1998 https://doi.org/10.1006/enrs.1998.3865
  48. Oda N, Sogawa CA, Sogawa N, Onodera K, Fyryta H, Yamamoto T: Metallothionein expression and localization in rat bone tissue after cadmium injection, Toxicol Lett 123 : 143 150, 2001 https://doi.org/10.1016/S0378-4274(01)00387-3
  49. Ogawa K, Oka K, Miyanishi T: Chitin and Chitosan and related enzymes, Lilcakis JP(Ed.), pp, 327 345, 1984
  50. Okuda H, Kato H, Tsujita T: Antihypertensive and antihyperlipemic actions of chitosan, Kor J Chitin Chitosan 2(3) : 49 59, 1997
  51. Ormrod OJ, Holmes CC, Miller TE: Dietary chitosan inhibits hypercholesterolaemia and atherogenesis in the apolipo-protein E deficient mouse model of atherosclerosis, Atherosclerosis 138 : 329 334, 1998 https://doi.org/10.1016/S0021-9150(98)00045-8
  52. Pannacciulli N, Cantatore FP, Minenna A, Bellacicco M, Giorgino R, De Pergola G: Urinary albwnin excretion is independently associated with Creactive protein levels in overweight and obese nondiabetic premenopausal women, J Intern Med 250(6) : 502 507, 2001 https://doi.org/10.1046/j.1365-2796.2001.00914.x
  53. Piscator M: Proteinuria in chronic cadmium poisoning, Arch Environ Health 12 : 345 359, 1966 https://doi.org/10.1080/00039896.1966.10664381
  54. Piscator M: Dietary exposure to cadmium and health effects: Impact of environmental changes, Environ Health Perspect 63: 127 131, 1985 https://doi.org/10.2307/3430038
  55. Prudden JF, Migel P, Hanson P, Friedrich L, Balassa L: The discovery of a potent pure chemical wound healing accelerator. Am J Surg 119(5) : 560 564, 1970 https://doi.org/10.1016/0002-9610(70)90175-3
  56. Ramirez DC, Gimenez MS: Lipid modification in mouse peritoneal macrophages after chronic cadmium exposure, Toxicology 172: 1 12, 2002 https://doi.org/10.1016/S0300-483X(01)00560-1
  57. Rhee SJ, Kim SO, Choe WK, Cho SH: Effect of cadmium dose injection on peroxidative damage in rat liver. J Kor Soc Food Nutr 21(6) : 601 607, 1992
  58. Shimizu M, Morita S: Effects of fasting on cadmium toxicity, glutathione metabolism, and metallothionein synthesis in rats, Toxicol Appl Pharmacol 103(1) : 28 39, 1990 https://doi.org/10.1016/0041-008X(90)90259-W
  59. Spagna G, Barbagallo RN, Casarini D: A novel chitosan derivative to immobilize alpha L rhamnopyranosidase from Aspergillus niger for application in beverage technologies, Enzyme Microb Technol28(4 5) : 427 438, 2001 https://doi.org/10.1016/S0141-0229(00)00340-9
  60. Springate JE, Van Lies JB, Noble B, Feld LG: Progressive glomerular injury after recovery from acute glomerulone-phritis in rats, Clin Immunol Immunopathol 61(3) : 309 319, 1991 https://doi.org/10.1016/S0090-1229(05)80003-0
  61. Squibb KS, Cousins RJ, Silbon BL, Levin S: Liver and intestinal metallothionein function in acute cadmium toxicity, Exp Mol Pathol 25: 163 171, 1976 https://doi.org/10.1016/0014-4800(76)90026-5
  62. Sugano M, Fujilcawa T, Hiratsuji Y, Nalcashima H, Fukuda N, Hasegawa Y: A novel use of chitosan as a hypocholesterolemic agent in rats, Am J Clin Nutr 33(4) : 787 793, 1980 https://doi.org/10.1093/ajcn/33.4.787
  63. Sunitha S, Nagaraj M, Varalalcshmi P: Hepatoprotective effect of lupeol and lupeollinoleate on tissue antioxidant defence system in cadmium induced hepatotoxicity in rats, Fitoterapia 72(5) : 516 523, 2001 https://doi.org/10.1016/S0367-326X(01)00259-3
  64. Suzuki K, Okawa Y, Hashimoto K, Suzuki S, Suzuki M: Protecting effect of chitin and chitosan on experimentally induced murine candidiasis, Microbiol Immunol 28(8) 903 912, 1984 https://doi.org/10.1111/j.1348-0421.1984.tb00746.x
  65. Suzuki KT, Uehara H, Takano Y, Aoki Y: Inhibition of pro-tein secretion from liver by cadmium, Biochem Pharmacol 34(23) : 4171 4173, 1985 https://doi.org/10.1016/0006-2952(85)90212-6
  66. Suzuki I(, Milcami T, Okawa Y, Tokoro A, Suzuki S, Suzuki M: Antitumor effect of hexa N acetylchitohexaose and chitohexaose, Carbohydr Res 151 : 403 408, 1986 https://doi.org/10.1016/S0008-6215(00)90359-8
  67. Suzuki K, Okawa Y, Suzuki S, Suzuki M: Candidacidal effect of peritoneal exudate cells in mice administered with chitin or chitosan: the role of serine protease on the mechanism of oxygen independent candidacidal effect, Microbiol Immumlol 31(4) : 375 379, 1987 https://doi.org/10.1111/j.1348-0421.1987.tb03098.x
  68. Taniguchi N, Tsukada Y, Hirai H: Acquirement of fetal pro-perties in hepatoma on glutathione metabolism, Biochim Biophys Acta 354: 161 167, 1974 https://doi.org/10.1016/0304-4165(74)90001-4
  69. Tokoro A, Tatewalci N, Suzuki K, Mikami T, Suzuki S, Suzuki M: Growth inhibitory effect of hexa N acetyl- chitohexaose and chitohexaose against Meth A solid tumor, Chern Pharm Bull 36(2) : 784 790, 1988 https://doi.org/10.1248/cpb.36.784
  70. Tong P, Baba Y, Adachi Y: Adsorption of metal ions on a new chelating Ion Exchange resin chemically derived from chitosan, Chern Lett 7 : 1529 1532, 1991
  71. Tor J, Pascual C, Segura RM, Vilaseca J, Schwartz S, Guardia J: Glucose phosphate isomerase and glutathione reductase in benign and malignant extrahepatic cholestasis, Clin Chern 27(4) : 634 635, 1981
  72. Waalkes MP, Hjelle JJ, Klaassen CD: Transient induction of hepatic metallothionein following oral ethanol administr- ation, Toxicol Appl Pharmacol 74(2) : 230 236, 1984 https://doi.org/10.1016/0041-008X(84)90147-9
  73. Webb M: Protection by zinc against cadmium toxicity, Biochern Pharmacol 21(20) : 2767 2771, 1972 https://doi.org/10.1016/0006-2952(72)90024-X
  74. Webster WS: Cadmium induced fetal growth retardation in the mouse, Arch Environ Health 33(1) : 36 42, 1978 https://doi.org/10.1080/00039896.1978.10667306