Lab Anim Res.  2011 Sep;27(3):227-234. 10.5625/lar.2011.27.3.227.

In vitro antioxidant properties of a ginseng intestinal metabolite IH-901

• Lee NJ
• Lee JW
• Sung JH
• Lee YJ
• Kang JK

• KMID: 1850050
• DOI: http://doi.org/10.5625/lar.2011.27.3.227

Abstract

IH-901 (20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol or compound K) is a final intestinal bacterial metabolite of ginseng in humans. It has various pharmacologic effects such as antiaging, immunopotentiation, antistress, and antimetastatic activities. We analyzed the antioxidant activities of IH-901 using several assays including: total antioxidant activity, reductive potential, 1,1-diphenyl-2-picryl-hydrazyl, hydroxyl, superoxide and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays, a nitric oxide scavenging assay and a lipid peroxidation assay. At concentrations of 25 and 100 microg/mL, IH-901 inhibited lipid peroxidation of a linoleic acid emulsion with a potency comparable to ascorbic acid and butylated hydroxyanisole. The reductive potential of IH-901 increased in a concentration-dependent manner. IH-901 exhibited strong DPPH, hydroxyl, superoxide and ABTS radical scavenging activities. IH-901 was also an effective inhibitor of lipid peroxidation, although IH-901 had only a mild scavenging activity against nitric oxide. These results suggest that IH-901 may be a useful antioxidant agent against reactive oxygen species.

Keyword

IH-901; ginseng intestinal metabolite; antioxidant activity

MeSH Terms

Ascorbic Acid
Benzothiazoles
Butylated Hydroxyanisole
Humans
Linoleic Acid
Lipid Peroxidation
Nitric Oxide
Panax
Reactive Oxygen Species
Sulfonic Acids
Superoxides
Ascorbic Acid
Benzothiazoles
Butylated Hydroxyanisole
Linoleic Acid
Nitric Oxide
Reactive Oxygen Species
Sulfonic Acids
Superoxides

Figure

• 

  Figure 1 Total antioxidant capacity and reductive potential of IH-901, BHA and ascorbic acid. For total antioxidant capacity (A), briefly, IH-901 or standard samples in 2.5 mL of potassium phosphate buffer were added to a linoleic acid emulsion in potassium phosphate buffer. The mixed solution was incubated at 37℃ in a glass flask in the dark, and the peroxide value was determined by reading the absorbance at 500 nm after reaction with FeCl2 and thiocyanate at intervals during the incubation. For reductive potential (B), briefly, the different concentrations of IH-901 (1.56-50 µg/mL) in 1 mL of distilled water were mixed with phosphate buffer and potassium ferricyanide. An aliquot (2.5 mL) of TCA was added to the mixture, which was then centrifuged for 10 min at 1000×g. The upper layer of the solution was mixed with distilled water and FeCl3, and the absorbance was measured at 700 nm. BHA: butylated hydroxyanisole, IH-901: ginseng intestinal metabolite-I.

• 

  Figure 2 DPPH (A), superoxide (B), hydroxyl (C) and ABTS (D) radical scavenging activities of BHA, ascorbic acid and IH-901. The radical scavenging ability of varying concentrations (3.125-100 µg/mL) of BHA, ascorbic acid and IH-901 was analyzed by measuring their inhibitory effects on the absorbance of DPPH, superoxide, hydroxyl and ABTS radicals. The results are expressed as mean±SD of the % inhibition of the absorbance of DPPH, superoxide, hydroxyl and ABTS radicals. DPPH: 1,1-diphenyl-2-picrylhydrazyl, ABTS: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), BHA: butylated hydroxyanisole, IH-901: ginseng intestinal metabolite-I.

• 

  Figure 3 Percentage inhibition of nitric oxide radicals (A) and lipid peroxidation (B) in the presence of different concentrations (3.125-1000 µg/mL) of ascorbic acid and IH-901. Nitric oxide (NO) radical scavenging activity was analyzed by measuring their inhibitory effects on the absorbance of the NO reaction product. It was analyzed by measuring inhibition of the absorbance of the thiobarbituric acid reaction product. Lipid peroxide production measured by UV detection of pink chromogens of malondialdehyde (MDA) in respective control reactions were 3.17±0.77. The results are expressed as mean±SD of the % inhibition of the absorbance of the NO and MDA. IH-901: ginseng intestinal metabolite-I.

Reference

1. Aruoma O. Free radicals, oxidative stress, and antioxidants in human health and disease. J Am Oil Chem Soc. 1998; 75(2):199–212.

2. Cerutti PA. Oxidant stress and carcinogenesis. Eur J Clin Invest. 1991; 21(1):1–5. PMID: 1907547.

3. Jung CH, Seog HM, Choi IW, Park MW, Cho HY. Antioxidant properties of various solvent extracts from wild ginseng leaves. Food Sci Technol-LEB. 2006; 39(3):266–274.

4. Halliwell B. Antioxidant characterization : Methodology and mechanism. Biochem Pharmacol. 1995; 49(10):1341–1348. PMID: 7763275.

5. Jung CH, Seog HM, Choi IW, Cho HY. Antioxidant activities of cultivated and wild Korean ginseng leaves. Food Chem. 2005; 92(3):535–540.

6. Kang KS, Yamabe N, Kim HY, Okamoto T, Sei Y, Yokozawa T. Increase in the free radical scavenging activities of American ginseng by heat processing and its safety evaluation. J Ethnopharmacol. 2007; 113(2):225–232. PMID: 17618072.

7. Hasegawa H, Sung JH, Huh JH. Ginseng intestinal bacterial metabolite IH901 as a new anti-metastatic agent. Arch Pharm Res. 1997; 20(6):539–544. PMID: 18982256.

8. Lee BH, Lee SJ, Hui JH, Lee S, Sung JH, Huh JD, Moon CK. In vitro antigenotoxic activity of novel ginseng saponin metabolites formed by intestinal bacteria. Planta Med. 1998; 64(6):500–503. PMID: 9741293.

9. Sung JH, Hasegawa H, Matsumiya S, Uchiyama M, Ha JY, Lee MS, Huh JD. Metabolism of ginseng saponins by human intestinal bacteria. Korean J Pharmacogn. 1995; 26:360–367.

10. Zhang D, Yasuda T, Yu Y, Zheng P, Kawabata T, Ma Y, Okada S. Ginseng extract scavenges hydroxyl radical and protects unsaturated fatty acids from decomposition caused by iron-mediated lipid peroxidation. Free Radic Biol Med. 1996; 20(1):145–150. PMID: 8903691.

11. Kang J, Lee Y, No K, Jung E, Sung J, Kim Y, Nam S. Ginseng intestinal metabolite-I (GIM-I) reduces doxorubicin toxicity in the mouse testis. Reprod Toxicol. 2002; 16(3):291–298. PMID: 12128103.

12. Wang T, Yu X, Qu S, Xu H, Han B, Sui D. Effect of ginsenoside Rb3 on myocardial injury and heart function impairment induced by isoproterenol in rats. Eur J Pharmacol. 2010; 636:121–125. PMID: 20371232.

13. Yokozawa T, Satoh A, Cho EJ. Ginsenoside-Rd attenuates oxidative damage related to aging in senescence-accelerated mice. J Pharm Pharmacol. 2004; 56(1):107–113. PMID: 14980007.

14. Yang J, Guo J, Yuan J. In vitro antioxidant properties of rutin. Food Sci Technol-LEB. 2008; 41(6):1060–1066.

15. Oyaizu M. Studies on product of browning reaction prepared from glucose amine. Jpn J Nutr. 1986; 44:307–315.

16. Liu F, Ooi VEC, Chang ST. Free radical scavenging activities of mushroom polysaccharide extracts. Life Sci. 1997; 60(10):763–771. PMID: 9064481.

17. Aruoma OI. Deoxyribose assay for detecting hydroxyl radicals. Methods Enzymol. 1994; 233:57–66.

18. Shimada K, Fujikawa K, Yahara K, Nakamura T. Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem. 1992; 40(6):945–948.

19. Shirwaikar A, Shirwaikar A, Rajendran K, Punitha IS. In vitro antioxidant studies on the benzyl tetra isoquinoline alkaloid berberine. Biol Pharm Bull. 2006; 29(9):1906–1910. PMID: 16946507.

20. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999; 26(9-10):1231–1237. PMID: 10381194.

21. Rai S, Wahile A, Mukherjee K, Saha BP, Mukherjee PK. Antioxidant activity of Nelumbo nucifera (sacred lotus) seeds. J Ethnopharmacol. 2006; 104(3):322–327. PMID: 16239089.

22. Halliwell B, Gutteridge JM. Free Radicals in Biology and Medicine. 1986. 2nd ed. Tokyo: Japan Scientific Societies Press;p. 42–53.

23. Gülçin I, Küfrevioglu ÖI, Oktay M, Büyüukokuroglu ME. Antioxidant, antimicrobial, antiulcer and analgesic activities of nettle (Urtica dioica L.). J Ethnopharmacol. 2004; 90(2-3):205–215. PMID: 15013182.

24. Gülçin I. Antioxidant activity of caffeic acid (3,4-dihydroxycinnamic acid). Toxicology. 2006; 217(2-3):213–220. PMID: 16243424.

25. Halliwell B, Gutteridge JM. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J. 1984; 219(1):1–14. PMID: 6326753.

26. Parejo I, Viladomat F, Bastida J, Rosas-Romero A, Flerlage N, Burillo J, Codina C. Comparison between the radical scavenging activity and antioxidant activity of six distilled and nondistilled mediterranean herbs and aromatic plants. J Agric Food Chem. 2002; 50(23):6882–6890. PMID: 12405792.

27. Feng T, Du Y, Li J, Hu Y, Kennedy JF. Enhancement of antioxidant activity of chitosan by irradiation. Carbohydr Polym. 2008; 73(1):126–132.

28. Razali N, Razab R, Junit SM, Aziz AA. Radical scavenging and reducing properties of extracts of cashew shoots (Anacardium occidentale). Food Chem. 2008; 111(1):38–44.

29. Govindarajan R, Rastogi S, Vijayakumar M, Shirwaikar A, Rawat AK, Mehrotra S, Pushpangadan P. Studies on the antioxidant activities of Desmodium gangeticum. Biol Pharm Bull. 2003; 26(10):1424–1427. PMID: 14519948.

30. Kim KT, Yoo KM, Lee JW, Eom SH, Hwang IK, Lee CY. Protective effect of steamed American ginseng (Panax quinquefolius L.) on V79-4 cells induced by oxidative stress. J Ethnopharmacol. 2007; 111(3):443–450. PMID: 17276636.