3-(4′-hydroxyl-3′, 5′-dimethoxyphenyl) Propionic Acid Suppresses NO Production and Elevates GSH Levels in Murine Macrophages

  • Song, Young-Sun (School of Food Science and Nutrition and BPRC, Inje University) ;
  • Choi, Chun-Yeon (School of Food Science and Nutrition and BPRC, Inje University) ;
  • Suh, Hongsuk (Department of Chemistry and Chemistry Institute for Functional Materials, Pusan National University) ;
  • Song, Yeong-Ok (Department of Food Science and Kimchi Research Institute, Pusan National University)
  • Published : 2004.09.01


Previous studies have shown that kimchi and kimchi-derived 3-(4'-hydroxyl-3', 5'-dimethoxyphenyl) propionic acid have anti-oxidative and hypolipidemic effects in rats and rabbits. This study was designed to investigate whether chemically synthesized 3-(4'-hydroxyl-3', 5' -dimethoxyphenyl) propionic acid (HDMPPA) may ameliorate oxidative stress through the regulation of nuclear factor KB (NFkB) activation in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage cells. Treatment of RAW 264.7 cells with 400 uM of HDMPPA significantly reduced LPS-stimulated nitric oxide (NO) production. Treatments with HDMPPA at 100 uM to 400 uM concentrations significantly elevated glutathione (GSH) level. However, cell viability and thiobarbituric acid-reactive substances (TBARS) concentrations were not affected by the concentrations of HDMPPA used. The specific DNA binding activities of NFKB, a transcription factor which is sensitive to oxidative stress, were not down-regulated by HDMPPA treatments. These results suggest that HDMPPA may have weak anti-oxidative activity against LPS challenge by scavenging NO and stimulating GSH production.


  1. Shen SC, Lee WR, Lin HY, Huang HC, Ko CH, Yang LL, Chen YC. 2002. In vitro and in vivo inhibitory activities of rutin, wogonin, and quercetin on lipopolysaccharide-induced nitric oxide and prostaglandin E(2) production. Eur J Pharmacol 446: 187-194
  2. Lee SK, Hong CH, Huh SK, Kim SS, Oh OJ, Min HY, Park KK, Chung WY, Hwang JK. 2002. Suppressive effect of natural sesquiterpenoids on inducible cyclooxygenase (COX-2) and nitric oxide synthase (iNOS) activity in mouse macrophage cells. J Environ Pathol Toxicol Oncol 21: 141-148
  3. Stolz DB, Zamora R, Vodovotz Y, Loughran PA, Billiar TR, Kim YM, Simmons RL, Watkins SC. 2002. Peroxisomal localization of inducible nitric oxide synthase in hepatocytes. Hepatology 36: 81-93
  4. Berliner JA, Navab M, Fogelman AM, Frank JS, Demer LL, Edwards PA, Watson AD, Lusis AJ. 1995. Atherosclerosis: Basic mechanisms: Oxidation, inflammation, and genetics. Circulation 91: 2488-2496
  5. Albrecht EW, Stegeman CA, Tiebosch AT, Tegzess AM, Van Goor H. 2002. Expression of inducible and endothelial nitric oxide synthases, formation of peroxynitrite and reactive oxygen spesies in human chronic renal transplant failure. Am J Transplant 2: 448-453
  6. Gobert AP, Mersey BD, Cheng Y, Blumberg DR, Newton JC, Wilson KT. 2002 Cutting edge: urease release by Helicobacter phlori stimulates macrophage inducible nitric oxide synthase. J Immunol 168: 6002-6006
  7. Iqbal M, Cohen RI, Marzouk K, Liu SF. 2002. Time course of nitric oxide, peroxynitrite, and antioxidants in the endotoxemic heart. Crit Care Med 30: 1291-1296
  8. Berendji D, Kolb-Bachofen V, Meyer KL, Kroncke KD. 1999. Influence of nitric oxide on the intracellular reduced glutathione pool: Different cellular capacities and strategies to encounter nitric oxide-mediated stress. Free Radic Biol Med 27: 773-780
  9. Bowie AG, Moynagh PN, O'Neill LA. 1997. Lipid peroxidation is involved in the activation of NF-?B by tumor necrosis factor but interleukin-1 in the human endothelial cell line ECV304. J Biol Chem 272: 25941-25950
  10. Schreck R, Meier B, Mannel DN, Droge W, Baeuerle PA. 1992. Dithiocarbamates as potent inbibitors of nuclear factor kappa B activation in intact cells. J Exp Med 175: 1181-1194
  11. Bonizzi G, Piette J, Schoonbroodt S, Greimers R, Havard L, Merville MP, Bours V. 1999. Reactive oxygen intermediate-dependent NF-$\kappa$B activation by interleukin-1$\beta$ requires 5-lipoxygenase or NADPH oxidase activity. Mol Cell Biol 19: 1950-1960
  12. Matthews JR, Wakasugi N, Virelizier JL, Yodoi J, Hay RT. 1992. Thioredoxin regulates the DNA binding activity of NF-kappaB by reduction of a disulfide bond involving cysteine 62. Nucleic Acids Res 20: 3821-3830
  13. Lo AH, Liang YC, Lin-Shiau SY, Ho CT, Lin JK. 2002. Carnosol, an antioxidant in rosemary, suppresses inducible nitric oxide synthase through down-regulating nuclear factor- ?B in mouse macrophages. Carcinogenesis 23: 983-991
  14. Wang J, Mazza G. 2002. Inhibitory effects of anthocyanins and other phenolic compounds on nitric oxide production in LPS/IFNγ-activated RAW264.7 macrophages. J Agric Food Chem 50: 850-857
  15. Prabhu KS, Zamamiri-Davis F, Stewart JB, Thomson JT, Sordillo LM, Reddy CC. 2002. Selenium deficiency increases the expression of inducible nitric oxide synthase in RAW 264.7 macrophage: role of nuclear factor-?B in up-regulation. Biochem J 366: 203-209
  16. Ippouchi K, Itou H, Azuma K, Higashio H. 2002. Effect of naturally occurring organosulfur compounds on nitric oxide production in lipopolysaccharide-activated macrophages. Life Sci 71: 411-419
  17. Victor VM, Guayerbas N, Puerto M, De la Fuente M. 2001. Changes in the ascorbic acid levels of peritoneal lymphocytes and macrophages of mice with endotoxin- induced oxidative stress. Fre Radic Res 35: 907-916
  18. Victor VM, De la Fuente M. 2002. N-acetylcysteine improves in vitro the function of macrophage from mice with endotoxin-induced oxidative stress. Free Radic Res 36: 33-45
  19. Kheir-Eldin AA, Motawi TK, Gad MZ, Abd-ElGawad HM. 2001. Protective effect of vitamin E, $\beta$-carotene and N-acetylcysteine from the brain oxidative stress induced in rats by lipopolysaccharide. Inter J Biodchem Cell Biol 33: 475-482
  20. Lee YM, Kwon MJ, Kim JK, Suh HS, Choi JS, Song YO. 2004. Isolation and identification of active principle in Chinese cabbage kimchi responsible for antioxidant effect. Korean J Food Sci Technol 36: 129-133
  21. Kim HJ, Kwon MJ, Seo JM, Kim JK, Song SH, Suh HS, Song YO. 2004. The effect of 3-(4-hydroxyl-3',5'-dimethoxyphenyl) propionic acid in Chinese cabbage kimchi on lowering hypercholesterolemia. J Korean Soc Food Sci Nutr 33: 52-58
  22. Kim HJ. 2004. The preventive and therapeutic effects of 3-(4'-hydroxyl-3',5'-dimethoxyphenyl) propionic acid, an active principle in Kimchi, on atherosclerosis in rabbits. PhD Dissertation. Pusan National University, Busan, Korea
  23. Fautz R, Husen B, Hechenberger C. 1001. Application of the neutral red assay (NR assay) to monolayer cultures of primary hepatocytes: rapid colorimetric viability determination for the unscheduled DNA synthesis test (UDS). Mutat Res 253: 173-179
  24. D'Agostino P, Ferlazzo V, Milano S, La Rosa M, Di Bella G, Caruso R, Barbera C, Grimaudo S, Tolomeo M, Feo S, Cillari E. 2001. Anti-inflammatory effects of chemically modified tetracyclines by the inhibition of nitric oxide and interleukin-12 synthesis in J774 cell line. International Immunopharmacology 1: 1765-1776
  25. Fraga CG, Leibovita RM, Roeder RG. 1988. Lipid peroxidation measured as thiobarbituric-reactive substances in tissue slices:characterization and comparision with homogenates and microsomes. Free Radic Biol Med 4: 155-161
  26. Tietze F. 1969. Enzymic method for quantitative determination of nanogram amounts of total and oxidaized glutathione: applications to mammalian blood and other tissue. Anal Biochem 27: 502-522
  27. Dignam JD, Lebovitz RM, Roeder RG. 1983. Accurate transcription initiation by RNA polymerase II in a soluble extract isolated from mammalian nuclei. Nucleic acids Res 11: 1475-1489
  28. Bradford MM. 1976. A rapid and sensitive method for the quantification of microgram quantities of proteins utilizing the principle of protein-dye binding. Ann Biochem 72: 248-254
  29. Yen GC, Lai HH. 2002. Inhibitory effects of isoflavones on nitric oxide-or peroxynitrite-mediated DNA damage in RAW 264.7 cells and $\Phi$X174DNA. Food and Chemical Toxicology 40: 1433-1440
  30. Mizutani K, Ikeda K, Nishikata T, Yamori Y. 2000. Phytoestrogens attenuate oxidative DNA damage in vascular smooth muscle cells from stroke-prone spontaneously hypertensive rats. J Hypertension 18: 1833-1840
  31. Ide N, Lau BHS. 2001. Garlic compounds minimize intracellular oxidative stress and inhibit nuclear factor-?B activation. J Nutr 131: 1020S-1026S