Protective Effect of Sasa borealis Leaf Extract on AAPH-Induced Oxidative Stress in LLC-PK1 Cells

  • Hwang, Ji-Young (Department of Food Science and Nutrition & Kimchi Research Institute) ;
  • Lee, Hee-Seob (Department of Food Science and Nutrition & Kimchi Research Institute) ;
  • Han, Ji-Sook (Department of Food Science and Nutrition & Kimchi Research Institute)
  • Received : 2010.11.11
  • Accepted : 2011.03.02
  • Published : 2011.03.31


This study was designed to investigate the protective effect of Sasa borealis leaf extract on 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress in LLC-PK1 cells (porcine kidney epithelial cells). The butanol fraction from Sasa borealis leaf extract (SBBF) was used in this study because it possessed strong antioxidant activity and high yield among fractions. Exposure of LLC-PK1 cells to 1 mM AAPH for 24 hr resulted in a significant decrease in cell viability, but SBBF treatment protected LLC-PK1 cells from AAPH-induced cell damage in a dose dependant manner. To determine the protective action of SBBF against AAPH-induced damage of LLC-PK1 cells, we measured the effects of SBBF on lipid peroxidation and antioxidant enzymes activities of AAPH treated cells as well as scavenging activities on superoxide anion radical and hydroxyl radical. SBBF had a protective effect against the AAPH-induced LLC-PK1 cellular damage and decreased lipid peroxidation and increased activities of antioxidant enzymes such as superoxide dismutase and glutathione peroxidase. Furthermore, SBBF showed strong scavenging activity against superoxide anion radical. The $IC_{50}$ value of SBBF was $28.45{\pm}1.28\;{\mu}g/mL$ for superoxide anion radical scavenging activity. The SBBF also had high hydroxyl radical scavenging activity ($IC_{50}=31.09{\pm}3.08\;{\mu}g/mL$). These results indicate that SBBF protects AAPH-induced LLC-PK1 cells damage by inhibiting lipid peroxidation, increasing antioxidant enzyme activities and scavenging free radicals.


  1. Xiangshu P, Xiang LP, Kim HY, Cho EJ. 2008. Antioxidative activity of geranium (Pelargonium inquinans Ait) and its active component, 1,2,3,4,6-penta-o-galloyl-$\beta$-Dglucose. Phytother Res 22: 534-538.
  2. Halliwell B. 1987. Oxidants and human disease: some new concepts. Faseb J 1: 358-364.
  3. Yokozawa T, Cho EJ, Hara Y, Kitani K. 2000. Antioxidative activity of green tea treated with radical initiator 2,2'-azobis(2-aminopropane) dihydrochloride. J Agric Food Chem 48: 5068-5073.
  4. Laguerrea M, Lecomtea J, Villeneuve P. 2007. Evaluation of the ability of antioxidants to counteract lipid oxidation: Existing methods, new trends and challenge. Prog Lipid Res 46: 244-282.
  5. Park HS, Lim JH, Kim HJ, Choi HJ, Lee IS. 2007.Antioxidant flavone glycosides from the leaves of Sasaborealis. Arch Pharm Res 30: 161-166.
  6. Znag Y, Ding XL. 1996. Studies on anti-oxidative fraction in bamboo leaves and its capacity to scavenge active oxygen radicals. J Bamboo Research 15: 17-24.
  7. Hwang JY, Han JS. 2007. Inhibitory effects of Sasa borealis leaves extracts on carbohydrate digestive enzymes and postprandial hyperglycemia. J Korean Soc Food Sci Nutr 36: 1391-1398.
  8. Hwang JY. 2009. Hypoglycemic and anti-glucotoxic effects of Sasa borealis. PhD Dissertation. Pusan National University, Busan, Korea. p 51-99.
  9. Yamaguchi T, Takamura H, Matoba T, Terao J. 1998.HPLC method for evaluation of the free radical-scavenging activity of food by using 1,1-diphenyl-2-picrylhydrazyl.Biosci Biotechnol Biochem 62: 1201-1204.
  10. Piao XL, Park IH, Baek SH, Kim HY, Park MK, ParkJH. 2004. Antioxidative activity of furanocoumarins isolated from Angelicae dahuricae. J Ethnopharmacol 93:243-246.
  11. Carmichael J, Degraff WG, Gazdar AF, Minna JD,Mitchell JB. 1987. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res 47: 936-942.
  12. Fraga CG, Leibovita RM, Roeder RG. 1988. Lipid peroxidation measured as thiobarbituric-reactive substances in tissue slices: characterization and comparison with homogenates and microsomes. Free Radic Biol Med 4: 155-161.
  13. Kirby AJ, Schmidt RJ. 1997. The antioxidant activity of Chinese herbs for eczema and placebo herbs-I. J Ethnopharmacol56: 103-108.
  14. Chung SK, Osawa T, Kawakishi S. 1997. Hydroxyl radical-scavenging effects of spices and scavengers from brown mustard (Brassica niger). Biosci Biotechnol Biochem61: 118-123.
  15. Bradford MM. 1976. A rapid and sensitive method for the quantification of microgram quantities of proteins utilizing the principle of protein-dye binding. Ann Biochem72: 248-254.
  16. Marklund S, Marklund G. 1974. Involvement of the superoxide anion radical in antioxidant of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem47: 469-474.
  17. Lawrence RA, Burk RF. 1976. Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun 71: 952-958.
  18. Piao XL, Kim HY, Yokozawa T, Lee YA, Piao XS, ChoEJ. 2005. Protective effects of broccoli (Brassica oleracea) and its active components against radical-induced oxidative damage. J Nutr Sci Vitaminol 51: 142-147.
  19. Yamamoto Y, Niki E, Eguchi J, Kamiya Y, ShimasakiH. 1985. Oxidation of biological membranes and its inhibition. Free radical chain oxidation of erythrocyte ghost membranes by oxygen. Biochim Biophys Acta 819: 29-36.
  20. Terao K, Niki E. 1986. Damage to biological tissues induced by radical initiator 2,2'-azobis(2-amidinopropane) dihydrochloride and its inhibition by chain-breaking antioxidants. Free Radic Biol Med 2: 193-201.
  21. Hochstein P, Jain SK. 1981. Association of lipid peroxidation and polymerization of membrane proteins with erythrocyte aging. Fed Proc 40: 183-188.
  22. Cho EJ, Yokozawa T, Rhyu DY, Mitsuma T, TerasawaK, Park JC. 2000. Protective activity from hydrophilic and lipophilic free radical generators of Wen-Pi-Tang and its crude drug extracts in LLC-PK1 cells. J Traditional Medicines 17: 245-252.
  23. Day BJ. 2004. Catalytic antioxidants: a radical approach to new therapeutics. Drug Discovery Today 9: 557-566.
  24. Halliwell B, Gutteridge JMC. 1999. Free radicals in biology and medicine. Oxford University Press, Oxford, UK.p 22-29.
  25. Jun Y, Limin W, Rosemary L, Edward G, Leonard M,Pike B. 2009. Antioxidant activity of citrus limonoids, flavonoids and coumarins. J Agric Food Chem 53: 2009-2014.
  26. Mitsuda H, Yasumoto K, Iwami K. 1966. Antioxidative action of indole compounds during the autoxidation of linoleic acid. Eiyo Shokuryo 19: 210-214.
  27. Park YO, Lim HS. 2009. Antioxidant activities of bamboo (Sasa borealis) leaf extract according to extraction solvent.J Korean Soc Food Sci Nutr 38: 1640-1648.
  28. Pokorny J. 1991. Natural antioxidants for food use. Trends Food Sci Technol 2: 223-227.
  29. Lui F, Ng TB. 1999. Antioxidative and free radical scavenging activities of selected medicinal herbs. J Life Sci 66: 725-735.
  30. Tanaka N, Wada H, Fuchino H, Inoue Y, Yasuda D,Kanada S, Kiyokawa C, Ashida N, Suzuki T, YamazakiK, Hakamatsuka N. 1998. Constituent of bamboos and bamboo grasses. Yakugaku Zasshi 118: 332-337.
  31. Park HS. 2006. Antioxidative compounds from the leaves of Sasa borealis. PhD Dissertation. Chonnam National University, Gwangju, Korea. p 20-21.
  32. Alptekin N, Seckin S, Dogru-Abbasoglu S, Yelkenci F,Kocak-Toker N, Toker G, Uysal M. 1996. Lipid peroxides,glutathione, $\gamma$-glutamylcysteine synthetase and $\gamma$-gutamyltranspeptidase activities in several tissues of rats following water-immersion stress. Pharmacol Res 34:167-169.

Cited by

  1. Antioxidant Activity and Inhibitory Effect on Oxidative DNA Damage of Ethyl Acetate Fractions Extracted from Cone of Red Pine (Pinus densiflora) vol.29, pp.2, 2016,
  2. Protective Effects of Phellinus linteus and Curry-Added Cooked Mixed Grain Rice Extracts on Oxidative Stress-Induced LLC-PK1 Cell Damage vol.43, pp.11, 2014,
  3. root vol.61, pp.2, 2018,