Toxicological Research

Korean Society of Toxicology & Korea Environmental Mutagen Society (한국독성학회)
권호 표지
DOI QR코드

DOI QR Code

Intragastrically Applicated CCl4-Thiopental Sodium Enhanced Lipid Peroxidation and Liver Fibrosis (Cirrhosis) in Rat: Malonedialdehyde as a Parameter of Lipid Peroxidation Correlated with Hydroxyproline as a Parameter of Collagen Synthesis (Deposition)

  • Kim, Ki-Young (Dept. of Pathology, Medical School and Professional Graduate School of Oriental Medicine) ;
  • Cho, Syung-Eun (Division of Bionanochemistry, Natural Science, Wonkwang University) ;
  • Yu, Byung-Soo (Division of Bionanochemistry, Natural Science, Wonkwang University)
  • Published : 2009.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

We investigated the pathogenesis of liver tissue damage during the lipid peroxidation and fibrogenesis with the observation of correlations between the parameters of collagen synthesis (and deposition) and lipid peroxidation in liver fibrosis (cirrhosis) rats. Rats were randomly divided into two groups, normal and $CCl_4$-thiopental sod. intoxicated group. And the one group was treated intragastrically with the mixture of $CCl_4$-thiopental sod. 3 times per week for 3 weeks. The liver tissue and sera were used for the measurement of hydroxyproline (HYP), malonedialdehyde (MDA) and superoxide dismutase (SOD). Biochemical parameters such as aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total-bilirubin and blood urea nitrogen (BUN) were measured. Additionally, the expression of collagen ${\alpha}1$(III) and $\beta$-actin mRNA was observed by RTPCR. The histological change in liver tissue was also observed by Masson's trichrome and H&E staining. Correlation analysis was carried by Spearman's rho method. All biochemical parameters except total-bilirubin were significantly higher in the $CCl_4$-thiopental sod. treated group than that of the normal group (p < 0.01). In the $CCl_4$-thiopental sod. treated group, Hyp as a parameter of collagen synthesis (deposition) and MDA as a metabolite of lipid peroxidation, were significantly elevated by 1.98 and 2.11 times higher than that of the normal group (p < 0.001) respectively. The activity of SOD in the $CCl_4$-thiopental sod. treated group is decreased significantly by 44.8% (p < 0.001). And collagen ${\alpha}1$(III) mRNA was more expressed in the $CCl_4$-thiopental sod. treated group than that of the normal group. However, the expression of $\beta$-actin mRNA is showed similar in both of groups. A good correlation was observed between the content of hyp and MDA concentration (r = 0.70, n = 40) in the two groups. And the correlation between the levels of hyp and SOD (r = -0.71, n = 25) is also reliable. However, no correlation were observed between MDA concentration and SOD (r = -0.40, n = 25) in the two groups. Elevated levels of MDA in $CCl_4$-thiopental sod. treated rats indicated enhancement of lipid peroxidation, which is accompanied by a decrease in SOD activity. Moreover, we could confirm that the parameters of collagen synthesis (and deposition) is in good correlation with the metabolite of lipid peroxidation (MDA) and the lipid peroxidation antagonizing enzyme (SOD). Hence, we propose that (1) lipid peroxidation and collagen synthesis (and deposition) could be enhanced by intragastrically application of $CCl_4$-thiopental sod. during a short terms. And (2) the intoxication of $CCl_4$-thiopental sod. could be used for monitoring of lipid peroxidation and collagen synthesis (and deposition) for test of antioxidant and antifibrotic agent.

Keywords

References

  1. Parola, M., Lenarduzzi, G., Robino, G., Albano, E., Poli, G. and Dianzano, M.U. (1996). On the role of lipid peroxidation in the pathogenesis of liver damage induced by longstanding cholestasis. Free Radic Biol. Med., 20, 351-359 https://doi.org/10.1016/0891-5849(96)02055-2
  2. Schuppan, D. (1990). Structure of the extracellular matrix in normal and fibrotic liver: collagens and glycoproteins. Seminars in Liver Diseases., 10, 1-10 https://doi.org/10.1055/s-2008-1040452
  3. Poli, G. and Parola, M. (1997). Oxidative damage and fibrogenesis. Free Radic Biol Med., 22, 287-305 https://doi.org/10.1016/S0891-5849(96)00327-9
  4. Slater, T.F. (1994). Free radical mechanisms in tissue injury. Biochemical. J., 222, 1-15
  5. Esterbauer, H., Schauer, R.J. and Zollner, H. (1992). Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol. Med., 11, 81-128 https://doi.org/10.1016/0891-5849(91)90192-6
  6. Cheeseman, K.H., Albano, E.F., Tomasi, A. and Slater, T.F. (1985). Biochemical studies on the metabolic activation of halogenated alkanes. Environ. Health Perspect, 64, 85-101 https://doi.org/10.2307/3430001
  7. Ruwart, M.J., Wilkinson, K.F., Rush, B.D., Kim, K.Y. and Schuppan, D. (1989). The intergrated value of serum procollagen type III peptide over time hepatic hydroxyproline content and stainable collagen in a model dietry cirrhosis in the rat. Hepatology, 10, 801-806 https://doi.org/10.1002/hep.1840100509
  8. Kamimura, S., Gall, K., Britton, R.S., Bacon, B.R., Triadafilopoulos, G. and Tsukamoto, H. (1992). Increased 4-hydrononenal levels in experimental alcoholic liver disease: association of lipid peroxidation with liver fibrogenesis. Hepatology, 116, 448-453
  9. Jamall, I.S., Finelli, V.N. and Que Hee, S.S. (1981). A single method to determine nanogram levels of 4-hydroxyproline in biological tissue. Anal. Biochem., 112, 70-75 https://doi.org/10.1016/0003-2697(81)90261-X
  10. Ohkawa, H., Ohishi, N. and Yagi, K. (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal. Biochem., 95, 351-358 https://doi.org/10.1016/0003-2697(79)90738-3
  11. Okado-Mastsumoto, A. and Fridovich, I. (2001). Assay of superoxide dismutase: cautions relevant to the use of cytochrome c, a sulfonated tetrazolium, and cyanide. Anal. Biochem., 298, 337-342 https://doi.org/10.1006/abio.2001.5385
  12. Kauschke, S.G., Knorr, A., Heke, M., Kohlmeyer, J., Schauer, M., Theiss, G., Waehler, R. and Burchardt, E.R. (1999). Two assay for measuring fibrosis: reverse transcriptasepolymerase chain reaction of collagen $\alpha$1(III) mRNA is an early predictor of subsequent collagen deposition while a novel serum N-terminal procollagen (III) propeptide assay reflects manifest fibrosis in carbon tetrachloride-treated rats. Anal. Biochem., 275, 131-40 https://doi.org/10.1006/abio.1999.4316
  13. MacDonald, G.A., Bridle, K.R., Ward, P.J., Walker, N.I., Houglum, K., George, D.K., Smith, J.L., Powell, L.W., Crawford, D.H. and Ramm, G.A. (2001). Lipid peroxidation in hepatic steatosis in humans is associated with hepatic fibrosis and occurs predominately in acinar zone 3. J. Gastro. Hepatol., 16, 599-606 https://doi.org/10.1046/j.1440-1746.2001.02445.x
  14. Vengerovskii, A.I., Baturina, N.O., Chuchalin, V.S. and Saratikov, A.S. (1996). Role of lipid peroxidation in the mechanism of proliferation of hepatic fibrous tissue in experimental chronic hepatitis. Patol. Fiziol. Eksp. Ter., 2, 37-9.15
  15. Gasso, M., Rubio, M., Varela, G., Cabre, M., Caballeria, J., Alonso, E., Deulofem, R., Camps, J., Gimenez, A., Pajares, M., Pares, A., Mato, J.M. and Rodes, J. (1996). Effects of S-adenosylmethionine on lipid peroxidation and liver fibrogenesis in carbon tetrachloride-induced cirrhosis. J. Hepatology, 25, 200-205 https://doi.org/10.1016/S0168-8278(96)80074-2
  16. Hernandez-Munoz, R., Diaz-Munoz, M., Lopez, V., Lopez-Barrera, F., Yanez, L., Vidrio, S., Aranda-Fraustro, A. and Chagoya de Sanchez, V. (1997). Balance between oxidative damage and proliferative potential in an experimental rat model of CCl4-induced cirrhosis: protective role of adenosine administration. Hepatology, 26, 1100-1110
  17. Hernandez-Munoz, R., Diaz-Munoz, M., Suarez-Cuenca, J.A., Trejo-Solis, C., Lopez, V., Sanchez-Sevilla, L., Yanez, L. and De Sanchez, V.C. (2001). Adenosine reverses a preestablished $CCl_{4}$-induced micronodular cirrhosis through enhancing collagenolytic activity and stimulating hepatocyte cell proliferation in rats. Hepatology, 34, 677-687 https://doi.org/10.1053/jhep.2001.27949
  18. Cabre, M., Camps, J., Paternian, J.L., Ferre, N. and Joven, J. (2000). Time-course of changes in hepatic lipid peroxidation and glutathione metabolism in rats with carbon tetrachloride-induced cirrhosis. Clin. Exp. Pharmacol. Phys., 27, 694-699 https://doi.org/10.1046/j.1440-1681.2000.03322.x
  19. Chen, M.F., Mo, L.R., Lin, R.C., Kuo, J.Y., Chang, K.K., Liao, C. and Lu, F.J. (1997). Increase of resting levels of superoxide anion in the whole blood of patients with decompensated liver cirrhosis. Free Radic. Biol. Med., 23, 672-679 https://doi.org/10.1016/S0891-5849(97)00057-9
  20. Paradis, V., Mathurin, P., Kollinger, M., Imbert-Bismut, F., Charlotte, F., Piton, A., Opolon, P., Holstege, A., Poynard, T. and Bedossa, P. (1997). In situ detection of lipid peroxidation in chronic hepatitis C: correlation with pathological features. J. Clin. Pathol., 50, 401-406 https://doi.org/10.1136/jcp.50.5.401
  21. Bacon, B.R. and Britton, R.S. (1990). The pathology of hepatic iron overload: a free radical-mediated process? Hepatology, 11, 127-137 https://doi.org/10.1002/hep.1840110122
  22. Niemelae, O., Parkkila, S., Ylae-Herttuala, S., Villanueva, J., Ruebner, B. and Halsted, C.H. (1995). Sequential acetaldehyde production, lipid peroxidation and fibrogenesis in minipig model of alcohol induced liver disease. Hepatology, 22, 1208-1214
  23. Vendemiale, G., Grattagliano, I., Caruso, M.L., Serviddio, G., Valentini, A.M., Pirrelli, M. and Altomare, E. (2001). Increased oxidative stress in dimethylnitrosamine-induced liver fibrosis in the rat: effect of N-acetylcystein and interferone interferone-a. Toxicol. Appl. Pharmacol., 175, 130-139 https://doi.org/10.1006/taap.2001.9234
  24. Nadkarni, G.D. and D'Souza, N.B. (1998). Hepatic antioxidant enzymes and lipid peroxidation in carbon tetrachloride-induced liver cirrhosis in rats. Biochem. Med. Meta. Biol., 40, 42-45
  25. Shiba, M., Shimizu, I., Yasuda, M., Ii, K. and Ito, S. (1998). Expression of type I and type III collagens during the course of dimethylnitrosamine-induced hepatic fibrosis in rats. Liver, 18, 196-204
  26. Lee, K.S., Buck, M., Houglum, K. and Chojkier, M. (1995). Activation of hepatic stellate cells by TGF alpha and collagen type I is mediated by oxidative stress through cmyb expression. J. Clin. Invest., 96, 2461-2468 https://doi.org/10.1172/JCI118304
  27. Maher, J.J., Tragarakis, C. and Gimenez, A. (1994). Malondialdehyde stimulates collagen production by hepatic lipocytes only upon activation in primary culture. Alcohol. Alcoholism., 29, 605-610
  28. Ljubuncic, P., Tanne, Z. and Bomzon, A. (2000). Evidence of a systemic phenomenon for oxidative stress in cholestatic liver disease. Gut., 47, 710-716 https://doi.org/10.1136/gut.47.5.710
  29. Rhoden, E.L., Pereira-Lima, L., Kalil, A.N., Lucas, M.L., Mauri, M., Menti, E., Rhoden, C.R., Pereira-Lima, J., Zettler, C.G. and Bello-Klein, A. Effects of ischemia and reperfusion on oxidative stress in hepatic cirrhosis induced by carbon tetrachloride in rats. Kobe. J. Med. Sci., 46, 171-180