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Anti-inflammatory and antioxidant activities of constituents isolated from Pueraria lobata roots

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Abstract

In order to evaluate the anti-inflammatory and antioxidant activities of Pueraria lobata roots and its active components, in vitro inhibitory activities against lipopolysaccharide (LPS)-induced nitric oxide (NO) production, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) protein expression, and tert-butylhydroperoxide (t-BHP)-induced reactive oxygen species (ROS) generation in RAW 264.7 cells, as well as in vitro scavenging activities against 1,1-diphenyl-2-picrylhydrazyl (DPPH), peroxynitrite (ONOO−), nitric oxide (NO·), superoxide anion (·O2 −) and total ROS, and inhibitory activities against ONOO−-mediated tyrosine nitration, were determined. Repeated column chromatography was performed to isolate four known compounds from the anti-inflammatory and antioxidant EtOAc fraction: daidzein; genistein; puerarin; (+)-puerarol B-2-O-glucopyranoside; four known compounds from the anti-inflammatory n-hexane fraction: lupenone; lupeol; puerarol; coumestrol; seven known compounds from the antioxidant n-BuOH fraction: allantoin; 3′-hydroxypuerarin; daidzein 8-C-apiosyl-(1→6)-glucoside; puerarin; genistin; 3′-methoxypuerarin; daidzin. Among these compounds, lupenone and lupeol reduced NO production, as well as iNOS and COX-2 protein levels in LPS-stimulated RAW 264.7 cells. Furthermore, lupeol showed significant inhibitory activity against intracellular ROS generation by t-BHP. Meanwhile, 3′-hydroxypuerarin showed marked ONOO−, NO·, total ROS scavenging activities, and weak ·O2 − scavenging activity, while 3′-methoxypuerarin showed ONOO− scavenging activity and weak NO· and O2 − scavenging activities, suggesting that existence of the 3′-hydroxyl group in puerarin plays an important role in the scavenging of ONOO−, NO·, and total ROS, as well as inhibiting the ONOO−-mediated tyrosine nitration mechanism. These results indicate that P. lobata roots and its constituents may be a useful therapeutic and preventive approach to various inflammatory diseases and oxidative stress-related disease.

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References

  • Arao, T., Kinjo, J., Nohara, T., and Isobe, R., Oleanene-type triterpene glycosides from Puerariae radix. IV. Six new saponins from Pueraria lobata. Chem. Pharm. Bull., 45, 362–366 (1997).

    Article  PubMed  CAS  Google Scholar 

  • Backhouse, N., Delporte, C., Givernau, M., Cassels, B. K., Valenzuela, A., and Speisky, H., Anti-inflammatory and antipyretic effects of boldine. Agents Actions, 42, 114–117 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Baek, M. R., Choi, Y. H., Yoo, D. S., Kim, M. R., Choi, S. U., Hong, K. S., Kim, Y. S., Kim, Y. K., Lee, K. R., and Ryu, S. Y., Anti-proliferative components in the roots extract from Pueraria thunbergiana. Korean J. Pharmacog., 40, 46–50 (2009).

    Google Scholar 

  • Beckman, J. S., Beckman, T. W., Chen, J., Marshall, P. A., and Freeman, B. A., Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc. Natl. Acad. Sci. U. S. A., 87, 1620–1624 (1990).

    Article  PubMed  CAS  Google Scholar 

  • Bickoff, E. M., Lyman, R. L., Livingston, A. L., and Booth, A. N., Characterization of coumestrol, a naturally occurring plant estrogen. J. Am. Chem. Soc., 80, 3969–3971 (1958).

    Article  CAS  Google Scholar 

  • Blois, M. S., Antioxidant determination by the use of a stable free radical. Nature, 181, 1199–1202 (1958).

    Article  CAS  Google Scholar 

  • Chen, A. F., Chen, D. D., Daiber, A., Faraci, F. M., Li, H., Rembold, C. M., and Laher, I., Free radical biology of the cardiovascular system. Clin. Sci. (Lond), 123, 73–91 (2012).

    Article  CAS  Google Scholar 

  • Cherdshewasart, W. and Sutjit, W., Correlation of antioxidant activity and major isoflavonoid contents of the phytoestrogen-rich Pueraria mirifica and Pueraria lobata tubers. Phytomedicine, 15, 38–43 (2008).

    Article  PubMed  CAS  Google Scholar 

  • Chueh, F. S., Chang, C. P., and Chio, C. C., Puerarin acts through brain serotonergic mechanisms to induce thermal effects. J. Pharmacol. Sci., 96, 420–427 (2004).

    Article  PubMed  CAS  Google Scholar 

  • Conner, E. M. and Grisham, M. B., Inflammation, free radicals and antioxidants. Nutrition, 12, 274–277 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Cos, P., De Bruyne, T., Apers, S., Vanden Berghe, D., Pieters, L., and Vlietinck, A., Phytoestrogens: recent developments. Planta Med., 69, 589–599 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Ding, H. Y., Chen, Y. P., Chang, W. L., and Lin, H. C., Isoflavonoids and but-2-enolides from the roots of Pueraria lobata. The Chinese Pharmaceutical Journal, 56, 31–35 (2004).

    CAS  Google Scholar 

  • Geetha, T. and Varalakshmi, P., Anti-inflammatory activity of lupeol and lupeol linoleate in rats. J. Ethnopharmacol., 76, 77–80 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Guerra, M. C., Speroni, E., Broccoli, M., Cangini, M., Pasini, P., Minghett, A., Crespi-Perellino, N., Mirasoli-Forti, G., and Paolini, M., Comparison between Chinese medical herb Pueraria lobata crude extract and its main isoflavone puerarin antioxidant properties and effect on rat liver CYP-catalyzed drug metabolism. Life Sci., 67, 2997–3006 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Halliwell, B. and Gutteridge, J. M. C., Free Radicals in Biology and Medicine. Oxford University Press, 3rd ed., Oxford, 617–783 (2002).

    Google Scholar 

  • Hasegawa, M., Flavonoids of various Prunus species VI. The flavonoids in the wood of Prunus aequinoctialis, P. nipponica, P. maximow and P. avium. J. Am. Chem. Soc., 79, 1738–1740 (1957).

    Article  CAS  Google Scholar 

  • Hirakura, K., Morita, M., Nakajima, K., Sugama, K., Takagi, K., Niitsu, K., Ikeya, Y., Maruno, M., and Okada, M., Phenolic glucosides from the root of Pueraria lobata. Phytochemistry, 46, 921–928 (1997).

    Article  CAS  Google Scholar 

  • Hudson, A. T. and Bentley, R., Stereospecific effects in cobalt (II1)-triethylenetetramine-N-methyl-(S)-alanine complexes. The isolation of isoflavonoids from bacteria. Chem. Commun., 14, 830–831 (1969).

    Google Scholar 

  • James, F. W., Frederick, A. F., Geraldine, J. F., and Harry, S. M., Isolation of allantoin and adenosine from the marine sponge Tethya aurantia. Comp. Biochem. Physiol. B, 70, 799–801 (1981).

    Article  Google Scholar 

  • Jiang, R., Lau, K., Lam, H., Yam, W., Leung, L., Choi, K., Waye, M., Mak, T. C. W., Woo, K., and Fung, K., A comparative study on aqueous root extracts of Pueraria thomsonii and Pueraria lobata by antioxidant assay and HPLC fingerprint analysis. J. Ethnopharmacol., 96, 133–138 (2005).

    Article  PubMed  Google Scholar 

  • Jun, M., Fu, H.-Y., Hong, J., Wan, X., Yang, C. S., and Ho, C.-T., Comparison of antioxidant activities of isoflavones from kudzu root (Pueraria lobata Ohwi). J. Food Sci., 68, 2117–2122 (2003).

    Article  CAS  Google Scholar 

  • Jun, M. R., Hong, J. K., Jeong, W. S., and Ho, C. T., Suppression of arachidonic acid metabolism and nitric oxide formation by kudzu isoflavones in murine macrophages. Mol. Nutr. Food Res., 49, 1154–1159 (2005).

    Article  PubMed  CAS  Google Scholar 

  • Kang, B. H., Ecological flora of Korean medicinal plants, Geobook, Seoul, (2008).

    Google Scholar 

  • Kang, K. A., Chae, S., Koh, Y. S., Kim, J. S., Lee, J. H., You, H. J., and Hyun, J. W., Protective effect of Puerariae radix on oxidative stress induced by hydrogen peroxide and streptozotocin. Biol. Pharm. Bull., 28, 1154–1160 (2005).

    Article  PubMed  CAS  Google Scholar 

  • Kaufman, P. B., Duke, J. A., Brielmann, H., and Hoyt, J. E., A comparative survey of legume plants as sources of the isoflavones, genistein and daidzein: implications for human nutrition and health. J. Altern. Complement. Med., 3, 7–12 (1997).

    Article  PubMed  CAS  Google Scholar 

  • Keung, W. M. and Vallee, B. L., Kudzu root: an ancient Chinese source of modern antidipsotropic agents. Phytochemistry, 47, 499–450 (1998).

    Article  PubMed  CAS  Google Scholar 

  • Kim, H. K., Cheon, B. S., Kim, Y. H., Kim, S. Y., and Kim, H. P., Effects of naturally occurring flavonoids on nitric oxide production in the macrophage cell line RAW 264.7 and their structure-activity relationships. Biochem. Pharmacol., 58, 759–765 (1999).

    Article  PubMed  CAS  Google Scholar 

  • Kim, J. Y., Jung, K. J., Choi, J. S., and Chung, H. Y., Hesperetin: a potent antioxidant against peroxynitrite. Free Radic. Res., 38, 761–769 (2004).

    Article  PubMed  CAS  Google Scholar 

  • Kinjo, J. E., Furusawa, J. I., Baba, J., Takeshita, T., Yamasaki, M., and Nohara, T., Studies on the constituents of Pueraria lobata. Isoflavonoids and related compounds in the roots and the voluble stems. Chem. Pharm. Bull., 35, 4846–4850 (1987).

    Article  CAS  Google Scholar 

  • Kinjo, J. E., Takeshita, T., Abe, Y., Terada, N., Yamashita, H., Yamasaki, M., Takeuchi, K., Murakami, K., Tomimatsu, T., and Nohara, T., Studies on the constituents of Pueraria lobata. IV. Chemical constituents in the flowers and the Leaves. Chem. Pharm. Bull., 36, 1174–1179 (1988a).

    Article  CAS  Google Scholar 

  • Kinjo, J. E., Takeshita, T., and Nohara, T., Studies on the constituents of Pueraria lobata. V. A trypotphan derivative from Pueraria Flos. Chem. Pharm. Bull., 36, 4171–4173 (1988b).

    Article  CAS  Google Scholar 

  • Kirakosyan, A., Kaufman, P. B., Warber, S., Bolling, S., Chang, S., and Duke, J. A., Quantification of major isoflavonoids and L-canavanine in several organs of Kudzu vine (Pueraria lobata) and in starch samples derived from Kudzu roots. Plant Sci., 164, 883–888 (2003).

    Article  CAS  Google Scholar 

  • Kobuchi, H., Virgili, F., and Packer, L., Assay of inducible form of nitric oxide synthase activity: effect of flavonoids and plant extracts. Methods Enzymol., 301, 504–513 (1999).

    Article  PubMed  CAS  Google Scholar 

  • Kooy, N. W., Royall, J. A., Ischiropoulos, H., and Beckman, J. S., Peroxynitrite-mediated oxidation of dihydrorhodamine 123. Free Radic. Biol. Med., 16, 149–156 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Lebel, C. P. and Bondy, S. C., Sensitive and rapid quantitation of oxygen reactive species formation in rat synaptosomes. Neurochem. Int., 17, 435–440 (1990).

    Article  PubMed  CAS  Google Scholar 

  • Lee, J. S., Supplementation of Pueraria radix water extract on changes of antioxidant enzymes and lipid profile in ethanol-treated rats. Clin. Chim. Acta, 347, 121–128 (2004).

    Article  PubMed  CAS  Google Scholar 

  • Lee, S. J., Baek, H. J., Lee, C. H., and Kim, H, P., Antiinflammatory activity of isoflavonoids from Pueraria Radix and biochanin a derivatives. Arch. Pharm. Res., 17, 31–35 (1994).

    Article  CAS  Google Scholar 

  • Lin, R. C., Guthrie, S., Xie, C. Y., Mai, K., Lee, D. Y., Lumeng, L., and Li, T. K., Isoflavonoid compounds extracts from Pueraria lobata suppress alcohol preference in a pharmacogenetic rat model of alcoholism. Alcohol. Clin. Exp. Res., 20, 659–663 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Lin, R. C. and Li, T. K., Effects of isoflavones on alcohol pharmacokinetics and alcohol-drinking behavior in rats. Am. J. Clin. Nutr., 68, 1512S–1515S (1998).

    PubMed  CAS  Google Scholar 

  • Madureira, A. M., Ascenso, J. R., Valdeira, L., Duarte, A., Frade, J. P., Freitas, G., and Ferreira, M. J., Evaluation of the antiviral and antimicrobial activities of triterpenes isolated from Euphorbia segetalis. Nat. Prod. Res., 17, 375–380 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Margareth, B. C. G. and Miranda, J. S., Biological activities of lupeol, International J. Biomedical and Pharmaceutical Sci. Global Science Books, (2009).

  • Mohammad, S., Lupeol, a novel anti-inflammatory and anticancer dietary triterpene. Cancer Lett., 285, 109–115 (2009).

    Article  Google Scholar 

  • Moreau, R. A., Whitaker, B. D., and Hicks, K. B., Phytosterols, phytostanols, and their conjugates in foods: structural diversity, quantitative analysis, and health-promoting uses. Prog. Lipid Res., 41, 457–500 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Mossman, T., Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxic assays. J. Immunol. Methods, 65, 55–63 (1983).

    Article  Google Scholar 

  • Na, M. K., An, R. B., Min, B. S., Lee, S. M., Kim, Y. H., and Bae, K. H., Chemical constituents from Sorbus commixta. Nat. Prod. Sci., 8, 62–65 (2002).

    CAS  Google Scholar 

  • Na, M. K., Kim, B. Y., Osada, H., and Ahn, J. S., Inhibition of protein tyrosine phosphatase 1B by lupeol and lupenone isolated from Sorbus commixt. J. Enzyme Inhib. Med. Chem., 24, 1056–1059 (2009).

    Article  PubMed  CAS  Google Scholar 

  • Nagaraj, M., Sunitha, S., and Varalakshmi, P., Effect of lupeol, a pentacyclic triterpene, on the lipid peroxidation and antioxidant status in rat kidney after chronic cadmium exposure. J. Appl. Toxicol., 20, 413–417 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Namgoong, S. Y., Lee, C. H., and Kim, H. P., Effects of isoflavonoids on mouse lymphocyte proliferation in vitro. Arch. Pharm. Res., 17, 236–239 (1994).

    Article  CAS  Google Scholar 

  • Nguemfo, E. L., Dimo, T., Dongmo, A. B., Azebaze, A. G., Alaoui, K., Asongalem, A. E., Cherrah, Y., and Kamtchouing, P., Anti-oxidative and anti-inflammatory activities of some isolated constituents from the stem bark of Allanblackia monticola Staner L.C (Guttiferae). Inflammopharmacology, 17, 37–41 (2009).

    Article  PubMed  CAS  Google Scholar 

  • Ohshima, Y., Okuyama, T., Takahashi, K., Takizawa, T., and Shibata, S., Isolation and high performance liquid chromatography (HPLC) of isoflavonoids from the Pueraria root. Planta Med., 54, 250–254 (1988).

    Article  PubMed  CAS  Google Scholar 

  • Park, J. S., Park, H. Y., Kim, D. H., Kim, D. H., and Kim, H. K., ortho-Dihydroxyisoflavone derivatives from aged Doenjang (Korean fermented soypaste) and its radical scavenging activity. Bioorg. Med. Chem. Lett., 18, 5006–5009 (2008).

    Article  PubMed  CAS  Google Scholar 

  • Qicheng, F., Some current study and research approaches relating to the use of plants in the traditional Chinese medicine. J. Ethnopharmacol., 2, 57–63 (1980).

    Article  PubMed  CAS  Google Scholar 

  • Rajic, A., Kweifio-Okai, G., Macrides, T., Sandeman, R. M., Chandler, D. S., and Polya, G. M., Inhibition of serine proteases by anti-inflammatory triterpenoids. Planta Med., 66, 206–210 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Ramirez Apan, A. A., Pérez-Castorena, A. L., and de Vivar, A. R., Anti-inflammatory constituents of Mortonia greggii Gray. Z. Naturforsch. C, 59, 237–243 (2004).

    PubMed  Google Scholar 

  • Rong, H., Stevens, J. F., Deinzer, M. L., Cooman, L. D., and Keukeleire, D. D., Identification of isoflavones in the roots of Pueraria lobata. Planta Med., 64, 620–627 (1998).

    Article  PubMed  CAS  Google Scholar 

  • Saleem, M., Afaq, F., Adhami, V. M., and Mukhtar, H., Lupeol modulates NF-κB and PI3K/Akt pathways and inhibits skin cancer in CD-1 mice. Oncogene, 23, 5203–5214 (2004).

    Article  PubMed  CAS  Google Scholar 

  • Sheu, F., Lai, H. H., and Yen, G. C., Suppression effect of soy isoflavones on nitric oxide production in RAW 264.7 macrophages. J. Agric. Food Chem., 49, 1767–1772 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Shibata, S., Murakami, T., and Nishikawa, Y., Studies on the constituents of Japanese and Chinese crude drugs. É. On the constituents of Pueraria root. J. Pharm. Soc. Japan, 79, 757–760 (1959).

    CAS  Google Scholar 

  • Shin, E. M., Zhou, H. Y., Guo, L. Y., Kim, J. A., Lee, S. H., Merfort, I., Kang, S. S., Kim, H. S., Kim, S., and Kim, Y. S., Anti-inflammatory effects of glycyrol isolated from Glycyrrhiza uralensis in LPS-stimulated RAW 264.7 macrophages. Int. Immunopharmacol., 8, 1524–1532 (2008).

    Article  PubMed  CAS  Google Scholar 

  • Sies, H., Klotz, L. O., Sharov, V. S., Assmann, A., and Briviba, K., Protection against peroxynitrite by selenoproteins. Z. Naturforsch. C, 53, 228–232 (1998).

    PubMed  CAS  Google Scholar 

  • Speroni, E., Guerra, M. C., Rossetti, A., Pozzetti, L., Sapone, A., Pasini, P., Roda, A., Paolini, M., and Cantelli-Forti, G., Antioxidant activity of Pueraria lobata (Willd.) in the rat. Phytother. Res., 10, s95–s97 (1996).

    Article  Google Scholar 

  • Valko, M., Free radicals and antioxidants in normal physiological functions and human disease. Int. J. Biochem. Cell, 39, 44–84 (2007).

    Article  CAS  Google Scholar 

  • Vallyathan, V. and Shi, X., The role of oxygen free radicals in occupational and environmental lung diseases. Environ. Health Perspect., 105, 165–177 (1997).

    PubMed  CAS  Google Scholar 

  • Wada, S., Iida, A., and Tanaka, R., Screening of triterpenoids isolated from Phyllanthus flexuosus for DNA topoisomerase inhibitory activity. J. Nat. Prod., 64, 1545–1547 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Wang, J. and Mazza, G., Inhibitory effects of anthocyanins and other phenolic compounds on nitric oxide production in LPS/IFN-γ-activated RAW 264.7 macrophages. J. Agric. Food Chem., 50, 850–857 (2002).

    Article  PubMed  CAS  Google Scholar 

  • You, Y. J., Nam, N. H., Kim, Y., Bae, K., and Ahn, B. Z., Antiangiogenic activity of lupeol from Bombax ceiba. Phytother. Res., 17, 341–344 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Ziegler, H. L., Staerk, D., Christensen, J., Hviid, L., Hagerstrand, H., and Jaroszewski, J. W., In vitro Plasmodium falciparum drug sensitivity assay: inhibition of parasite growth by incorporation of stomatocytogenic amphiphiles into the erythrocyte membrane. Antimicrob. Agents Chemother., 46, 1441–1446 (2002).

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Basic Herbal Medicine Research Group, Korea Institute of Oriental Medicine, Daejeon, 305-811, Korea

    Seong Eun Jin

  2. Department of Food and Life Science, Pukyong National University, Busan, 608-737, Korea

    You Kyung Son & Jae Sue Choi

  3. College of Pharmacy, Catholic University of Daegu, Daegu, 712-702, Korea

    Byung-Sun Min

  4. Department of Food Science and Human Nutrition, Chonbuk National University, Jeonju, 561-756, Korea

    Hyun Ah Jung

  5. Blue-Bio Industry RIC, Dongeui University, Busan, 614-714, Korea

    Jae Sue Choi

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Jin, S.E., Son, Y.K., Min, BS. et al. Anti-inflammatory and antioxidant activities of constituents isolated from Pueraria lobata roots. Arch. Pharm. Res. 35, 823–837 (2012). https://doi.org/10.1007/s12272-012-0508-x

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