Anti-Oxidative and Anti-Inflammatory Effects of Cheongajihwang-Tang Extract on RAW264.7 Cells

청아지황탕(靑蛾地黃湯)의 RAW264.7 Cell에서 항산화 및 항염증 효능 연구

  • Yoon, Jeong-won (Department of Korean Rehabilitation Medicine, College of Korean Medicine, Se-Myung University) ;
  • Kim, Soon-Joong (Department of Korean Rehabilitation Medicine, College of Korean Medicine, Se-Myung University) ;
  • Park, Dong-Su (Department of Korean Rehabilitation Medicine, College of Korean Medicine, Se-Myung University)
  • 윤정원 (세명대학교 한의과대학 한방재활의학과교실) ;
  • 김순중 (세명대학교 한의과대학 한방재활의학과교실) ;
  • 박동수 (세명대학교 한의과대학 한방재활의학과교실)
  • Received : 2016.06.23
  • Accepted : 2016.07.06
  • Published : 2016.07.31


Objectives This study was designed to investigate whether the Cheongajihwang-Tang (CT) has an inhibitory effect association with oxidation or inflammation in RAW264.7 cells. Methods Cytotoxic activity of CT extract on RAW264.7 cells was evaluated by using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) solution. Nitric oxide production was measured using Griess reagent system. The total phenolic contents and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was measured to evaluate the anti-oxidative effects of CT. Dichlorofluorescin diacetate (DCFH-DA) has been used as a substrate for measuring intracellular oxidant production. Results Cheongajihwang-Tang does not impair the cell viability in tested concentration. CT showed anti-oxidative effects in vitro by decreasing electron donating ability, and also showed anti-inflammatory effects suppressing NO and ROS expression in LPS induced RAW264.7 activation. CT inhibited the generation of intracellular ROS production as dose dependant manner. Conclusions CT has anti-oxidative effects and anti-inflammatory activities. These results indicate that CT extract has an anti-inflammatory activities via anti-oxidative effects.


  1. Bae GC, Bae DY. The anti-inflammatory effects of ethanol extract of Allium Hookeri cultivated in South Korea. Kor.J.Herbology. 2012;27(6):55-61.
  2. Greig FH, Kennedy S, Spickett CM. Physiological effects of oxidized phospholipids and their cellular signaling mechanisms in inflammation. Free Radic Biol Med. 2012;52(2):266-80.
  3. Lee DU. Oxidative stress and age-related changes in microsomal mixed function oxidase activity. Kor.J.Gerontol. 1991;1(2):187-201.
  4. Volicer L, Crino PB. Involvement of free radicals in dementia of the Alzheimer's type, a hypothesis. Neurobiol Aging. 1990;11:567-71.
  5. Kim HJ, Jin CB, Lee YS. Anti-oxidative activities of phenolic compound isolated from Inonotus obliquus. Kor.J.Pharmacognogy. 2007;38:1-16.
  6. Finkel T, Holbrook NJ. Oxidants, oxidative stress and the biology of aging. Nature. 2000;408(6809):239-47.
  7. Chung HY, Kim HJ, Kim JW. The inflammation hypothesis of aging : Molecular modulation by calorie restriction. Ann.N.Y Acad Sc. 2001;928:327-35.
  8. Kim HJ, Ahn MS, Kim GH, Kang MH. Antioxidant and antimicrobial activity of Pleurotus eryngii extracts prepared from different aerial part. Kor.J.Food Sci Technol. 2006;38:799-804.
  9. Kim YH. Cheonggangeuigam. Seoul:Seongbosa. 2001:304.
  10. Jin SM. Taepyeonghyeminhwajegukbang. Beijing:Ren min wei sheng Publishing INC. 1985:175.
  11. Qian Yi. Soayakjeungjikgyeol. Seoul:Yeogang Publishing INC. 2002:11.
  12. Kim SH, Jeong JC. Peroxynitrite scavenging activity and its mechanism of Cheonga-hwan. Kor.J.oriental medicine. 2002;23(4):55-63.
  13. Moon SS, Kim BS, Kang JS. Study on antioxidant action of Yukmijihwang-tang. Kor.J.oriental physiology&pathology. 2003;17(2):436-442.
  14. Jeong JC. Increased antioxidant enzyme activities and scavenging effects of oxygen free radicals by Cheongahwan. Journal of Korean Medicine. 1997;18(2):355-65.
  15. Seo YE, Lee EA, Bae HS, Shin MK, Hong MC. Antioxidant effects of the Herbs Composing Yukmijihwang-tang on PC12 cell. Kor.J.Oriental Physiology&Pathology. 2003;17(1):203-8.
  16. Lee GH, Yoo DY. Evaluation of Anti-Inflammatory Effects of Yukmijihwangtang and Individual Drug Substances Based on the Extraction Methods. The Journal of oriental obstetrics&gynecology. 2012;25(2):89-107.
  17. Kim MS, Seo BI, Kwak MA, Jee SY. Effect of Chungajihwangtang on osteoporosis in ovariectomized rats. Kor.J.Herbology. 2003;18(2):49-58.
  18. Velioglu YS, Mazza G, Gao L, Oomah BD. Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. J Agric Food Chem. 1998;46(10):4113-7.
  19. Kandaswami CE. Middleton E Jr. Free Radical Scavenging and Antioxidant Activity of Plant Flavonoids. Adv Exp Med Biol. 1994;366:351-76.
  20. Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of radiosensitivity. Cancer Res. 1987;47(4):943-6.
  21. Gheldof N, Engeseth NJ. Antioxidant capacity of honeys from various floral sources based on the determination of oxygen radical absorbance capacity and inhibition of in vitro lipoprotein oxidation in human serum samples. J Agric Food Chem. 2002;50(10):3050-5.
  22. Choi BK, Jeong SY, Park GS, Cho JH. Reactive Oxygen Substances and Diseases. Shinil Publishing INC. 2004;1:7-15.
  23. Park SN. Skin aging and antioxidants. Kor.J.Soc Cosmetic Chem. 1997;23:75-132.
  24. Stadtman ER, Berlett BS. Reactive oxygenmediated protein oxidation in aging and disease. Drug Metab Rev. 1998;30(2):325-43.
  25. Virag L, Szabo E, Gergely P and Szabo C. Peroxynitrite induced cytotoxicity: mechanism and opportunities for intervention. Toxicol Lett. 2003;140(1):113-124.
  26. Omata N, Tsukahara H, Ito S, Ohshima Y, Yasutomi M, Yamada A, Jiang M, Hiraoka M, Nambu M, Deguchi Y, Mayumi M. Increased oxidative stress in childhood atopic dermatitis. Life Science. 2001;69:223-8.
  27. Knight JA. The biochemistry of aging. Adv Clin Chem. 2000;35:1-62.
  28. Goldberg I. Functional foods. New York:Chapman&Hall Press. 1994:3-550.
  29. Valko M, Rhodes CJ, Moncol J, Izakovic M, Mazur M. Free radicals, metals and antioxidants in oxidative stressinduced cancer. Chem Biol Interact. 2006;160:1-40.
  30. Kim MJ, Chu WM, Park EJ. Antioxidant and antigenotoxic effects of shiitake mushrooms affected by different drying methods. Kor.J.Soc Food Sci Nutr. 2012;41:1041-8.
  31. Hwang TY. Bangyakhappyeon. Seoul:Namsandang. 1989:216-7.
  32. Hong WS. Huangjenaegyeongsomun. Seoul:Institute of Traditional Culture. 1993;19-20. 46.
  33. Lee ES, Ju HK, Moon TC, Lee E, Jahng Y, Lee SH, Son JK, Baek SH, Chang HW. Inhibition of nitric oxide and tumor necrosis factor-${\alpha}$ production by propenone compound through blockade of nuclear factor (NF)-kB activation in cultured murine macrophage. Biol Pharm Bull. 2004;27:617-20.
  34. Lin CH, Yeh CH, Lin LJ, Wang SD, Wang JS, Kao ST. Immunomodulatory effect of chinese herbal medicine formula sheng-fei-yu-chuan-tang in lipopolysaccharide-induced acute lung injury mice. Evid Based Complement Alternat Med. 2013;2013:976342.
  35. Mathiak G, Grass G, Herzmann T, Luebke T, Zetina CC, Boehm SA, Bohlen H, Neville LF, Hoelscher AH. Caspase-1-inhibitor ac-YVAD-cmk reduces LPS lethality in rats without affecting haematology or cytokine responses. Br J Pharmacol. 2000;131(3):383-6.
  36. Dai J, Mumper RJ. Plant phenolics: Extraction, analysis and their antioxidant and anticancer properties. Molecules. 2010;15:7313-52.
  37. Labuza TP. Kinetic of lipid oxidation in foods. CRS critical Rev. Food Technol. 1973;335.
  38. Meenakshi S, Umayaparvathi S, Arumugam M, Balasubramanian T. In vitro antioxidant properties and FTIR analysis of two seaweeds of Gulf of Mannar. Asian Pac J Trop Med 2012;66-70.
  39. Bredt DS, Snyder SH. Nitric oxide, a novel neuronal messenger. Neuron. 1992;8:3-11.
  40. Ryu, JH. Inhibitory activity of plant extracts on nitric oxide synthesis in LPS- activated macrophage. Phytother Res. 2003;17:485-9.
  41. Fiers W, Beyaert R, Declercq W, Vandenabeele P. More than one way to die: apoptosis, necrosis and reactive oxygen damage. Oncogene. 1999;18(54):7719-30.
  42. Choi WH, Oh YS, Ahn JY, Kim SR, Ha TY. Antioxidative and Protective Effects of Ulmus davidiana var. japonica Extracts on Glutamate-Induced Cytotoxicity in PC 12 Cells. Kor.J.Food Sci Technol. 2005;37(3):479-83.