Browse > Article

The Screening of Antioxidant and Antifibrotic Effect from Water Extracts of Herbs(LH) in Biliary Liver Fibrosis (Cirrhosis) Induced Rsts  

임진아 (원광대학교 한의학전문대학원)
김기영 (원광대학교 한의학전문대학원)
Publication Information
YAKHAK HOEJI / v.46, no.2, 2002 , pp. 129-136 More about this Journal
Abstract
The pathogenesis of cholestatic liver injury as well as the modulation of hepatic fibrogenesis is causally associated with involvement of reactive oxygen species (ROS) and free radical reactions. In this study, we investigated whether dried extracts of oriental medicine (LH) have antioxidant and antifibrotic effect under the biliary liver fibrosis (cirrhosis) c ondition. The female Sprague-Dawley rats were divided in six groups (Normal, N-LH, op-2, op-4, opLH-2, opLH-4) and were observed in 2 weeks or 4 weeks. For this purpose the rats were operated by bile duct ligation/scission (BDL/S), which induced to liver fibrosis and cirrhosis. After surgery, the prepared LH was administered p.o. 2 mι/day/rat in 2 weeks or 4 weeks for opLH groups. During the observation period, jaundices appeared in eyes, ears and tail of all BDL/S operated rats. And at the time of sacrifice, cholestasis was observed in proximal bile duct, especially the color of bile juice and urine in opLH-4 group showed more clear than op-2, op-4 and opLH-2 group. The value of clinical parameters and product of lipid peroxidation (MDA) in sera and the hydroxyproline (hyp) content in liver tissue were significantly increased in all liver fibrosis (cirrhosis) developed rats (p＜0.001~0.05). Among the clinical parameters of sera, value of BUN, ALP in opLH-4 group showed significantly lower than in op-4 group (p＜0.05, p＜0.001). The content of hyp in opLH-2, opLH-4 group (478.0 $\pm$ 134.3 $\mu\textrm{g}$/g 897.5 $\pm$ 118.2 $\mu\textrm{g}$/g) showed lower than in op-2, op-4 group (528.9 $\pm$ 220.7 $\mu\textrm{g}$/g, 1023.8 $\pm$ 277.1 $\mu\textrm{g}$/g) and then the value of MDA in opLH-4 was also significantly reduced to 59.4% of that in op-4 group (p＜0.001). The histological change (bile duct proliferation, fibrosis, collagen bundle) was similarly observed in op-2 group and in opLH-2 group, but the weak fibrosis and bile duct proliferation were observed in opLH-4 group compared with in op-4 group. Our data indicate that the 4 weeks treatment with LH extract suppressed lipid peroxidation and inhibited fibrotic (cirrhosis) process, and experimental cholestatic liver disease is associated with increased lipid peroxidation in BDL/S operated rats. Hence we concluded that the measurement of MDA and hyp can be useful monitor for the screening of antioxidant and antifibrotic effect in experimental liver fibrosis (cirrhosis), and LH has been shown to have hepatoprotective effect, antifibrotic effect and antioxidant effect.
Keywords
Liver fibrosis; lipid peroxidation; MDA; antioxidant ＆ antifibrotic effect;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Ljubuncic, P., Tanne, Z., Bomzon, A. : Evidence of a systemic phenomenon for oxidative stress in cholestatic liver disease. Gut 47(5), 710 (2000)
2 Pawlak, W., Kedziora, J., Zolynski, K., Kedziora-Kornatowska, K., Blaszczyk, J., Witkowski, P., Zieleniewski, J.: Effect of long term bed rest in men on enzymatic antioxidative defence and lipid peroxidation in erythrocytes. J. Gravit. Physiol. 5(1), 163 (1998)
3 Ono, M., Sekiya, C., Ohhira, M., Ohhira, M., Namiki, M., Endo, Y., Suzuki, K., Matsuda, Y., Taniguchi, N. : Elevated level of serum Mn-superoxide dismutase in patients with primary biliary cirrhosis: Possible involvement of free radicals in the pathogenesis in primary biliary cirrhosis. J. Lab. Clin. Med. 118(5), 476 (1991)
4 Friedman, S. L., Roll, J. F., Boyles, J., Bissel, D. M. : Hepatic lipocytes: The principal collagen production cells of normal rat liver. Proc. Natl. Acad. Sci. 82, 8681 (1985)
5 Zavoico, G. B. : New targets for the treatment of pathologic fibrosis, an unmet clinical need. Durg & Market Development 10(1), 2 (1999)
6 Gressner, A. M. and Bachem, M. G. : Cellular sources of noncollagenous matrix proteins: Role of fat-storing cells in fibrogenesis. Semin. Liver Dis. 10, 30 (1990)
7 Das, D., Pemberton, P. W., Burrows, P. C., Gordon, C., Smith, A., McMahon, R. F., Warnes, T. W. : Antioxidant properties of colchicine in acute carbon tetrachloride induced rat liver injury and its role in the resolution of established cirrhosis. Biochem. Biophys, Acta. 1502(3), 351 (2000)
8 Lin, C. C., Huang, P. C., Lin, J. M. : Antioxidant and hepatoprotective effects of Anoectochilus formosanus and Gynostemma pentaphyllum. Am. J. Clin. Med. 28(1), 87 (2000)
9 Parola, M., Leonarduzzi, G., Biasi, F., Albano, E., Biocca, M. E., Poli, G., Dianzani, M. U. : Vitamin E dietary supplementation protects against carbon tetrachloride-induced chronic liver damage and cirrhosis. Hepatology 16, 1014 (1992)
10 Nordmann, R., Ribiere, C., Rouach, H. : Implication of free radical mechanisms in ethanol-induced cellular injury. Free Radic. Biol. Med. 12, 219 (1992)   DOI   ScienceOn
11 Poli, G., Parola, M., Leonarduzzi, G., Pinzani, M. : Modulation of hepatic fibrogenesis by antioxidants. Mol. Aspects. Med. 14, 259 (1993)   DOI   ScienceOn
12 Bacon, B. R., Britton, R. S. : The pathology of hepatic iron overload: A free radical-mediated process? Hepatology 11, 127 (1990)
13 Cotran-Humar, R. ed. : Robbins pathologic basis of disease. 4th ed. 918 (1989)
14 Sokol, R. J., Deveraux, M., Mierau, G. W., Hambidge, K. M., Shikes, R. H. : Oxidant injury to hepatic mitochondrial lipids in rats with dietary copper overload: Modification by vitamin E deficiency. Gastroenterology 99, 1061 (1990)
15 정재열, 임진아, 박선영, 서의석, 제갈승주, 김기영. : 실험동물에서 복합천연물(DW)의 지질과산화 억제와 간섬유화 저해 효과검색. 약학회지 45, 513 (2001)
16 Gerok, W. ed. : Hepatologie. Urban & Schwarzenberg 547 (1987)
17 French, S. W., Wong, K., Albano, E., Hagbjork, A. L., Ingelman-Sundberg, M. : Effect of ethanol on cytochrom P450 2E1(CYP2E1), lipid peroxidation and serum protein adduct formation in relation to liver pathology pathogenesis. Exp. Mol. Pathol. 58, 61 (1993)   DOI   ScienceOn
18 Kose, K., Yazici, C., Assioglu, O. : The evaluation of lipid peroxidation and adenosine deaminase activity in patients with Behcet's disease. Clin. Biochem. 34(2), 125 (2001)
19 Tsai, L. Y., Lee, K. T., Tsai, S. M., Lee, S. C., Yu, H. S. : Change of lipid peroxide levels in blood and liver tissue of patients with obstructive jaundice. Clin. Chim. Acta. 215, 41 (1993)   DOI   ScienceOn
20 Slater, T. F. : Free radical mechanisms in tissue injury. Biochem. J. 222, 1 (1994)
21 Albano, E., Clot, P., Tabone, M., Arico, S., Ingelman-Sundberg, M. In., Poli, G., Albano, E., Dianzani, M. U., eds. : Free radicals: From basic science to medicine. Basel: Birkhauser Verlag. 310 (1993)
22 Niemela, O., Parkkila, S., Yla-Herttuala, S., Villanueva, J., Ruebner, B., Halsted, C. H. : Squential acetaldehyde production, lipid peroxidation, and fibrogenesis in a micropig model of alcohol-induced liver disease. Hepatology 22, 1208 (1995)
23 Halliwell, B. : Reactive oxygen species in living system; source, biochemistry and role in human disease. Am. Med. suppl 3C, 14 (1991)
24 Vaziri, N, D., Ni, Z., Oveisi, F., Liang, K., Pandian, R. : Enhanced nitric oxide inactivation and protein nitration by reactive oxygen species in renal insufficiency. Hypertension 39(1), 135 (2002)
25 Shimizu, I., Ma, Y. R., Mizobuchi, Y., Liu, F., Miura, T., Nakai, Y., Yasuda, M., Shiba, M., Horie, T., Amagaya, S., Kawada, N., Hori, H., Ito, S. : Effects of Sho-saiko-to, a Japanese herbal medicine on hepatic fibrosis in rats. Hepatology 29, 149 (1999)
26 Dianzani, M. U. ; Lipid peroxidation in ethanol poisoning; A critical reconsideration. Alcohol Acohol 20, 161 (1985)
27 Sies, H. ed. : Oxidative stress. Orlando. Academic Press (1985)
28 Deems, R. O., Skypala, P. L., Martinez-Hernandez, A., Friedman, L. S., Friedman, M. I. : Dietary fat exacerbates liver disease in bile duct-ligated rats. J. Nutr. 123, 1414 (1993)
29 Vendemiale, G., Grattagliano, I., Caruso, M. L., Serviddio, G., Valentini, A. M., Pirrelli, M., Altomare, E. : Increased oxidative stress in dimethylnitrosamine-induced liver fibrosis in the rats: Effect of N-acetylcysteine and interferon-$\alpha$. Toxicol. Appl. Pharm, 175, 130 (2001)
30 Ohkawa, H., Ohishi, N., Yagi, K : Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal. Biochem. 95, 351 (1979)   DOI   ScienceOn
31 Ljubuncic, P., Tanne, Z., Bomzon, A. : Ursodeoxycholic acid suppresses extent of lipid peroxidation in diseased liver in experimental cholestatic liver disease. Dig. Dis. Sci. 45(10), 1921 (2000)
32 Parola, M., Pinzani, M., Casini, A., Albano, E., Poli, G., Gentilini, A., Gentilini, P., Dianzani, M. U. : Stimulation of lipid peroxidation or 4-hydroxynonenal treatment increases procollagen alpha 1(I) gene expression in human liver fat-storing cells. Biochem. Biophys. Res. Commun. 194, 1044 (1993)   DOI   PUBMED   ScienceOn
33 Slater, T. F. ed. : Free radical mechanisms in tissue injury. London. Pion Ltd (1972)
34 Fabris, C., Pirisi, M., Panozzo, M. P., Soardo, G., Toniutto, P., Hocza, V., Bartoli, E. : Intensity of inflarnrnatory damage and serum lipid peroxide concentrations in liver disease. J Clin Pathol. 46, 364 (1993)   DOI   ScienceOn
35 Panozzo, M. P., Basso, D., Balint, L., Biasin, M. R., Bonvicini, P., Metus, P., Plebani, M. : Altered lipid peroxidation glutathione ratio in experimental extrahepatic cholestasis. Clin. Exp. Pharmacol. Physiol. 22, 266 (1995)   DOI   ScienceOn
36 Peres, W., Tunon, M. J., Collado, P. S., Herrmann, S., Marroni, N., Gonzalez-Gallego, J. : The flavonoid quercetin ameliorates liver damage in rats with biliary obstruction. J. Hepatol. 33(5), 742 (2000)
37 Parola, M., Muraca, R., Dianzani, I., Barrera, G., Leonarduzzi, G., Bendinelli, P., Piccoletti, R., Poli, G. : Vitamin E dietary supplementation inhibits transforming growth factor ${\beta}1$ gene expression in the rat liver. FEBS Lett. 308, 267 (1992)   DOI   PUBMED   ScienceOn
38 Bomzon, A., Holt, S., Moore, K. : Bile acid, oxidative stress and renal function in biliary obstruction. Semin. Nephrol, 17, 549 (1997)
39 Chen, M. F., Mo, L. R, Lin, R. C., Kuo, J.. Y., Chang, K K., Liao, C., Lu, F. J. : Increase of resting levels of superoxide anion in the whole blood of patients with decompensated liver cirrhosis. Free Radic. Biol. Med. 23, 672 (1997)   DOI   ScienceOn
40 Maher, J. J., Tzagarakis, C., Gimenez, A. : Malondealdehyde stimulates collagen production by hepatic lipocytes only upon activation in primary culture. Alcohol & Alcoholism 29, 605 (1994)
41 Muriel, P., Suarez, O. R. : Role of lipid peroxidation in biliary obstruction in rat. J. Appl. Toxicol. 14, 423 (1995)
42 Pancewicz, S. A., Skrzydlewska, E., Hermanowska-Szpakowicz, T., Zajkowska, J. M., Kondrusik, M. : Role of reactive oxygen species (ROS) in patients with erythema migrans, an early manifestation of Lyme borreliosis. Med. Sci. Monit. 7(6), 1230 (2001)
43 Fabris, C., Pirisi, M., Panozzo, M. P., Soardo, G., Toniutto, P., Hocza, V., Bartoli, E. : Intensity of inflammatory damage and serum lipid peroxide concentrations in liver disease. J. Clin. Pathol. 46(4), 364 (1993)
44 Sanchez, A., Alvarez, A. M., Benito, M., Fanregat, I. : Apoptosis induced by transforming growth factor-${\beta}$ in fetal hepatocyte primary cultures. J. Biol. Chem. 271, 7416 (1996)   DOI   PUBMED
45 Kamimura, S., Gall, K., Britton, R S., Triadafilopoulos, G., Tsukamoto, H. : Increased 4-hydroxynonenal levels in experimental alcoholic liver disease: Association of lipid peroxidation with liver fibrogenesis. Hepatology 16, 448 (1992)
46 Sokol, R. J., Deveraux, M., Khandwala, R. A. : Effect of dietary lipid and vitamin E on mito-chondrial lipid peroxidation and hepatic injury in the bile duct-ligated rat. J. Lipid Res. 32, 1349 (1991)
47 Lee, K. S., Buck, M., Houglum, K., Chojkier, M. : Activation of hepatic stellate cells by TGF${\alpha}$ and collagen type I is mediated by oxidative stress through c-myb expression. J. Clin. Invest. 96, 2461 (1995)   DOI   ScienceOn
48 Bedossa, P., Houghlum, K., Trautwein, C., Holstege, A., Chojkier, M. : Stimulation of collagen 1(I) gene expression is associated with lipid peroxidation in hepatocellular injury: A link to tissue fibrosis? Hepatology 19, 1262 (1994)
49 Herrera, B., Fernandez, M., Alvarez, A. M., Roncero, C., Benito, M., Gil, J., Fabregat, I. : Activation of caspases occurs downstream from radical oxygen species production, Bcl-xL down-regulation, and early cytochrome C release in apoptosis induced by transforming growth factor beta in rat fetal hepatocytes. Hepatology 34(3), 548 (2001)
50 Bedossa, P., Houblum, K., Trautwein, C., Holstege, A., Chojkier, M. : Stimulation of collagen alpha 1(I) gene expression is associated with lipid peroxidation in hepatocellular injury: A link to tissue fibrosis? Hepatology 19(5), 1262 (1994)
51 Parola, M., Leonarduzzi, G., Robino, G., Albano, E., Poli, G., Dianzani, M. U. : On the role of lipid peroxidation in the pathogenesis of liver damage induced by long-standing cholestasis. Free Radie. Biol. Med. 20, 351 (1996)   DOI   ScienceOn
52 Luper, S. : A review of plants used in the treatment of liver disease: Part 1. Altern. Med. Rev. 3(6), 410 (1998)
53 Mourelle, M., Muriel, P., Favari,L.,Franco, T. : Prevention of $CCl_4$-induced liver cirrhosis by silymarin. Fundam. Clin. Pharmacol. 3 183 (1989)
54 Singh S., Shackleton, G., Ah-Singh, E., Chakraborty, J., Bailey, M. E. : Antioxidant defenses in the bile duct-ligated rat. Gastroenterology 103, 1625 (1992)
55 Chen, M. F., Mo, L. R., Lin, R. C., Kuo, J. Y., Chang, K. K., Liao, C., Lu, F. J. : Increase of resting levels of superoxide anion in the whole blood of patients with decompensated liver cirrhosis. Free Radic. Biol. Med. 23(4), 672 (1997)
56 Friedman, S. L. : Cellular sources of collagen and regulation of collagen production in liver. Semin. Liver Dis. 10, 20 (1990)   DOI   PUBMED   ScienceOn
57 Kountras, J., Billing, B. H., Scheuder, P.J. : Prolonged bile duct obstruction: A new experimental model for cirrhosis in the rat. Br. J. Exp. Path. 65, 305 (1984)
58 Tsai, L. Y., Lee, K. T., Liu, T. Z. : Evidence for accelerated generation of hydroxyl radicals in experimental obstructive jaundices of rats. Free Radic. Biol. Med. 24, 732 (1998)   DOI   ScienceOn
59 Shinh, S., Shackleton, G., Ah-Sing, E., Chakraborty, J., Bailey, M. E. : Antioxidant defenses in the bile duct ligated rat. Gastroenterology 103, 1625 (1992)
60 Jamall, I. S. and Finelli, V. N. : A single method to determine nanogram levels of 4-hydroxyproline in biological tissue. Anal. Biochem. 112, 70 (1981)   DOI   ScienceOn
61 Poli, G. and Parola, M. : Oxidative damage and fibrogenesis. Free Radic. Biol. Med. 22, 287 (1997)   DOI   ScienceOn