DOI QR코드

DOI QR Code

The Effects of Selenium on Cadmium-Induced Toxicity and Lipid Peroxidation in Rat Hepatocyte Primary Culture

랫드 간세포 일차배양에서 셀레늄이 카드뮴에 의해 유도된 독성 및 지질과산화에 미치는 영향

  • Rhim, Tae-Jin (Department of Biotechnology, College of Life Science and Natural Resources, Sangji University)
  • 임태진 (상지대학교 생명자원과학대학 생명공학과)
  • Published : 2003.06.30

Abstract

The objective of present study was to investigate the antioxidative and hepatoprotective effects of selenium on cadmium-induced toxicity and lipid peroxidation in rat hepatocyte primary culture. To do this, two separate experiments were conducted. In Experiment 1, primary cultures of rat hepatocytes were incubated for 6 hr in the presence of various concentrations (1, 10, 50, 100, and $500\;{\mu}M$) of cadmium chloride. Cytotoxicity and lipid peroxidation were evaluated using the MTT assay and TBARS assay, respectively. Antioxidative and hepatoprotective effects were determined by measuring the activity of GOT and GSH-Px, respectively. Cell viability was reduced and lipid peroxidation was increased by cadmium in dose-dependent manners. There was significantly negative correlation (r=-0.943, p<0.01) between cell viability and lipid peroxidation GOT activity was increased and GSH-Px activity was decreased by cadmium at the concentration of $50\;{\mu}M$. In Experiment 2, primary cultures of rat hepatocytes were incubated for 6hr in the presence of 100\;{\mu}M$ of cadmium chloride and various concentrations (0.01, 0.1 and 1 ppm) of sodium selenite to assess the effect of selenium on cadmium-induced toxicity and lipid peroxidation. Cell viability and GSH-Px activity were increased by sodium selenite at the concentration of 1 ppm Whereas, lipid peroxidation and GOT activity were reduced by 0.1 ppm of sodium selenite. These results demonstrate that selenium has an antioxidative and hepatoprotective potentials against cadmium.

본 연구의 목적은 랫드 간세포 일차배양에서 카드뮴에 의해 유발된 세포독성 및 지질과산화에 대한 셀레늄의 항산화 및 간보호 효과를 조사하기 위함이다. 이를 위해 2개의 실험을 수행하였다. 실험 1에서는, 1, 10, 50, 100 및 $50\;{\mu}M$의 다양한 농도의 카드뮴으로 6시간 동안 간세포를 일차 배양하였다. 간세포 생존율과 지질과산화는 각각 MTT 방법과 TBARS 방법으로 측정하였다. 항산화 효과와 간보호 효과는 각각 GOT와 GSH-Px 활성을 측정함으로써 결정하였다. 카드뮴은 농도 의존적으로 세포 생존율을 감소시켰으며 지질과산화는 증가시켰다. 세포 생존율과 지질과산화 간에 유의적인 음의 상관관계(r=-0.943, p<0.01)가 관찰되었다. 카드뮴은 $50\;{\mu}M$ 농도에서 GOT의 활성을 증가시켰으나 GSH-Px의 활성은 감소시켰다. 실험 2에서는 셀레늄이 카드뮴에 의해 유발된 독성 및 지질과산화에 대한 효과를 연구하기 위해 $100\;{\mu}M$의 카드뮴과 0.01, 0.1 및 1 ppm의 다양한 농도의 셀레늄으로 6시간 동안 간세포 일차 배양하였다. 세포 생존율과 GSH-Px의 활성은 1 ppm의 셀레늄에 의해 증가되었다. 반면에, 지질과산화와 GOT의 활성은 0.1 ppm의 셀레늄에 의해 감소되었다. 이러한 연구 결과는 셀레늄이 카드뮴에 대한 항산화 및 보호 효과를 나타내 보이고 있다.

Keywords

References

  1. Samarawickrama, G. P. (1979) Biological effects of cadmium in mammals, In Webb M(ed):'The chemistry, biochemistry and biology of cadmium.', Elsevier-North Holland Biomedical Press, Amsterdam, p.341-422
  2. Rajanna, B., Hobson, M, Reese, J., Sample, E. and Chapatwala, K. D. (1984) Chronic hepatic and renal toxicity by cadmium in rats, Drug Chem. Toxicol. 7, 229-241 https://doi.org/10.3109/01480548409035105
  3. Kunimoto, M, Miysaka, K. and Miura, T. (1986) Changes in membrane properties of rat blood cells induced by cadmium accumulating in the membrane fraction J. Bioehem. Tokyo 99,3 97-406
  4. Chapatwala, K. D, Rajanna, B. and Desaiah, D. (1980) Cadmium-induced changes in gluconeogenic enzymes in rat kidney and liver, Drug Chem. Toxicol. 3, 407-420 https://doi.org/10.3109/01480548009030129
  5. Manca, D., Ricard, A. C, Trottier, B. and Chevalier, G. (1991) Studies on lipid peroxidation in rat tissues following administration of low and moderate doses of cadmium chloride, Toxicology 67, 303-323 https://doi.org/10.1016/0300-483X(91)90030-5
  6. Sarkar, S., Yadav, P., Trivedi, R., Bansal, A. K. and Bhatnagar, D. (1995) Cadmium-induced lipid peroxidation and the status of the antioxidant system in rat tissues, J. Trace Element Med. Biol. 9, 144-149 https://doi.org/10.1016/S0946-672X(11)80038-6
  7. Dalton, T., Fu, K., Enders, G. C., Palmiter, R. D. and Andrews, G. K. (1996) Analysis of the effects of overexpression of metallothionein-I in transgenic mice on the reproductive toxicology of cadmium, Environ. Health Persp. 104, 68-76 https://doi.org/10.2307/3432762
  8. Combs, G F. Jr. and Combs, S. B. (1984) The nutritional biochemistry of selenium, Ann. Rev. Nutr. 4, 257-280 https://doi.org/10.1146/annurev.nu.04.070184.001353
  9. Jamall I. S. and Smith, J. C. (1985) Effects of cadmium treatment on selenium-dependent and selenium-independent glutathione peroxidase activities and lipid peroxidation in the kidney and liver of rats maintained on various level of dietary selenium Toxicology 647, 102-105
  10. Olsson, U. (1986) Selenium deficiency and detoxication functions in the rat: Short-term effects of cadmium, Drug-Nutr. Interact. 4, 309-319
  11. Ognjanovic, B., Zikic, R. V., Stajn A, Saicic, Z. S., Kostic, M. M. and Petrovic, V. M. (1995) The effects of selenium on the antioxidant defense system in the liver of rats exposed to cadmium Physiol. Res. 44, 293-300
  12. Sugawara, N. and Sugawara, C. (1984) Selenium protection against testicular lipid peroxidation from cadmium J. Appl. Biochem. 6, 199-204
  13. Yiin, S. J. Chem C. L., Sheu, J. Y. and Lin, T. H. (1999) Cadmium induced lipid peroxidation in rat testes and protection by selenium Biometals 12, 353-359 https://doi.org/10.1023/A:1009277121164
  14. Yiin S. J., Chem, C. L., Sheu, J. Y., Tseng. W. C. and Lin T. H. (1999) Cadmium-induced renal lipid peroxidation in rats and protection by selenium, J. Toxicol. Env. Health 57, 101-111
  15. Seglen, P. O. (1976) Preparation of Isolated rat liver cell, Methods Cell Bioi. 13. 29-83 https://doi.org/10.1016/S0091-679X(08)61797-5
  16. Reitman, S. and Frankel. S. (1957) A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases, Am. J. Clin. Pathol. 28, 56-63
  17. Lii, C. K., Wang. S. T., Chen, H. W. and Sheen, L. Y. (1996) Glutathione and glutathione-related enzyme activities of male and female rat hepatocytes under various culture conditions, Arch. Toxicol. 70, 822-829 https://doi.org/10.1007/s002040050345
  18. lawrence, R. A. and Burk, R. F. (1976) Glutatione peroxidase activity in selenium deficient rat liver, Biochem. Biophys. Res. Commun. 71, 952-958 https://doi.org/10.1016/0006-291X(76)90747-6
  19. Uchiyama, M. and Mihara, M. (1978) Determination of malonaldehyde precursor in tissues by thiobarbituric acid test, Anal. Biochem. 86, 271-278 https://doi.org/10.1016/0003-2697(78)90342-1
  20. Foretz, M, Foufelle, F. and Ferre, P. (1999) Polyunsaturated fatty acids inhibit fatty acid synthase and spot-14-protein gene expression in cultured rat hepatocytes by a peroxidative mechanism Biochem. J. 341, 371-376 https://doi.org/10.1042/0264-6021:3410371
  21. Mosmann, T. (1983) Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays, J. Immunol. Methods 65, 55-63 https://doi.org/10.1016/0022-1759(83)90303-4
  22. Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem. 72, 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  23. Steel, R. G. D. and Torre, J. H. (1980) Principles and Procedures of Statistics, 2nd ed, McGraw-Hill, New York, p.186-187
  24. Fariss, M. W. (1991) Cadmium toxicity: unique cytoprotective properties of alpha tocopheryl succinate in hepatocytes, Toxicology 69, 63-77 https://doi.org/10.1016/0300-483X(91)90154-S
  25. Pourahmad J. and O'Brien, P. J. (2000) A comparison of hepatocyte cytotoxic mechamisms for $Cu^{2+}$ and $Cd^{2+}$, Toxicology 143, 263-272 https://doi.org/10.1016/S0300-483X(99)00178-X
  26. Stacey, N. H., Cantilena, Jr. L. R. and Klaassen, C. D. (1980) Cadmium toxicity and lipid peroxidation in isolated hepatocytes, Toxicol. Appl. Pharmacol. 53, 470-480 https://doi.org/10.1016/0041-008X(80)90359-2
  27. Muller, L. and Ohnesorge, F. K. (1982) Different response of liver parenchymal cells from starved and fed rats to cadmum, Toxicology 25, 141-150 https://doi.org/10.1016/0300-483X(82)90025-7
  28. Muller, L. (1986) Consequences of cadmium toxicity in rat hepatocytes: Effects of cadmium of the glutathione-peroxidase system Toxicol. Lett. 30, 259-265 https://doi.org/10.1016/0378-4274(86)90164-5
  29. Omaye, S. T. and Tappel A. L. (1975) Effect of cadmium chloride on the testicular soluble selenoenzyme, glutathione peroxidase, Res. Commun. Chern. Pathol. Pharmacol. 12, 695-711
  30. Splittgerber, A. G. and Tappel A. L. (1979) Inhibition of glutathione peroxidase by cadmium and other metal ions, Arch. Biochem. Biophys. 197, 534-542 https://doi.org/10.1016/0003-9861(79)90277-7
  31. Black, R. S., Whanger, P. D. and Tripp, M. J. (1979) Influence of silver, mercury, lead, cadmium and selenium on glutathione peroxidase and trasferase activities in rats, Bioi. Trace Element Res. 1, 313-324 https://doi.org/10.1007/BF02778833
  32. Pool M. L. (1981) Exposure and health effects of cadmium, Part 3. Effects of cadmium on enzyme activities, Tox. Enviorn. Chem. Rev. 4, 179-203
  33. Styblo, M and Thomas, D. J. (2001) Selenium modifies the metabolism and toxicity of arsenic in primary rat hepatocytes, Toxicol. Appl. Pharmacol. 172, 52-61 https://doi.org/10.1006/taap.2001.9134
  34. Othman, A. I. and EI Missiry, M. A. (1998) Role of selenium against lead toxicity in male rats, J. Biochem. mol. Toxicol. 12, 345-349 https://doi.org/10.1002/(SICI)1099-0461(1998)12:6<345::AID-JBT4>3.0.CO;2-V
  35. Laws, J., Latshaw, J. and Biggert, M. (1986) Selenium bioavailability in foods and feeds, Nutr. Rep. Int. 33, 13-24
  36. Greeder, G. A. and Milner, J. A. (1980) Factors influencing the inhibitory effect of selenium on mice inoculated with Ehrlich ascites tumor cells, Science 209, 825-827 https://doi.org/10.1126/science.7406957

Cited by

  1. Evaluation of Antioxidantive and Antimicrobial Activities of Garlic Stem and Red Cabbage, and Their Application to Pork Patties during Refrigerated Storage vol.30, pp.2, 2010, https://doi.org/10.5851/kosfa.2010.30.2.291
  2. Current research trends for heavy metals of agricultural soils and crop uptake in Korea vol.31, pp.1, 2012, https://doi.org/10.5338/KJEA.2012.31.1.75