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Quantitative analysis of the anti-inflammatory activity of orengedokuto II: berberine is responsible for the inhibition of NO production

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Abstract

Orengedokuto is a Kampo formula that has been used for removing “heat” and “poison” to treat inflammation, hypertension, gastrointestinal disorders, and liver and cerebrovascular diseases. We report here our analysis of the anti-inflammatory effect of the component crude drugs of orengedokuto and their constituents using the inhibition of nitric oxide (NO) production in the murine macrophage-like cell line J774.1. An initial comparison of NO production inhibitory activities of the extracts of the component crude drugs and their combinations revealed that the activity could be attributed to Phellodendron Bark and Coptis Rhizome. Berberine (1), the major constituent of these crude drugs, showed potent activity (IC50 4.73 ± 1.46 μM). Quantitative analysis of 1 in the extracts of all combinations of component crude drugs revealed that the amount of 1 in each extract of the combination of Scutellaria Root with either Phellodendron Bark and/or Coptis Rhizome was lower than that in the corresponding mixtures of the extracts of the individual crude drugs and that 1 was present in the precipitates formed during the decoction process. To the contrary, the differences in the amounts of 1 were smaller in the extracts containing Gardenia Fruit. These results indicated that the constituents of Scutellaria Root precipitated with 1 and that the constituents of Gardenia Fruit dissolved the precipitates. To identify the constituents affecting the solubility of 1, we fractionated the hot-water extracts of Scutellaria Root based on solubility tests of 1 to give baicalin (2), wogonin (3) and oroxyloside (4), which formed precipitates with 1.

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References

  1. The Ministry of Health, Labour and Welfare of Japan (2016) The Japanese pharmacopoeia. 17th edn. The Ministry of Health, Labour and Welfare of Japan, Tokyo, pp 1752–1754

  2. Cao YP, Gao C, Sun JH, Wang JZ, Zhou Q, Liu GQ (1996) Study on pharmacological effects of Huanglianjiedu Tang extract. J China Pharm Univ 27:605–608

    Google Scholar 

  3. Yoon SB, Lee YJ, Park SK, Kim HC, Bae H, Kim HM, Ko SG, Choi HY, Oh MS, Park W (2009) Anti-inflammatory effects of Scutellaria baicalensis water extract on LPS-activated RAW264.7 macrophages. J Ethnopharmacol 125:286–290

    Article  Google Scholar 

  4. Chen JJ, Huang CC, Chan HY, Li PY, Liang YC, Deng JS, Huang SS, Huang GJ (2017) Scutellaria baicalensis ameliorates acute lung injury by suppressing inflammation in vitro and in vivo. Am J Chin Med 45:137–157

    Article  Google Scholar 

  5. Li J, Liu Y, Chen X, Ding X, Wu S, Xie W (2006) Anti-endotoxin effects of baicalin extracted from Scutellariae Radix. Yiyao Daobao 25:1237–1240

    CAS  Google Scholar 

  6. Chi YS, Cheon BS, Kim HP (2001) Effect of wogonin, a plant flavone from Scutellariae Radix, on the suppression of cyclooxygenase-2 and the induction of inducible nitric oxide synthase in lipopolysaccharide-treated RAW264.7 cells. Biochem Pharmacol 61:1195–1203

    Article  CAS  Google Scholar 

  7. Chi YS, Kim HP (2005) Suppression of cyclooxygenase-2 expression of skin fibroblasts by wogonin, a plant flavone from Scutellariae Radix. Prostaglandins Leukot Essent Fatty Acids 72:59–66

    Article  CAS  Google Scholar 

  8. Lin WH, Kuo HH, Ho LH, Tseng ML, Siao AC, Hung CT, Jeng KC, Hou CW (2015) Gardenia jasminoides extracts and gallic acid inhibit lipopolysaccharide-induced inflammation by suppression of JNK2/1 signaling pathways in BV-2 cells. Iran J Basic Med Sci 18:555–562

    PubMed  PubMed Central  Google Scholar 

  9. Mao YF, Li YQ, Zong L, You XM, Lin FQ, Jiang L (2010) Methanol extract of Phellodendri Cortex alleviates lipopolysaccharide-induced acute airway inflammation in mice. Immunopharmacol Immunotoxicol 32:110–115

    Article  CAS  Google Scholar 

  10. Park YK, Chung YS, Kim YS, Kwon OY, Joh TH (2007) Inhibition of gene expression and production of iNOS and TNF-α in LPS-stimulated microglia by methanol extract of Phellondendri cortex. Int Immunopharmacol 7:955–962

    Article  CAS  Google Scholar 

  11. Iizuka N, Miyamoto K, Hazama S, Yoshimura K, Okita K, Fukumoto T, Yamamoto S, Tangoku A, Oka M (2000) Anticachectic effects of Coptidis Rhizoma, an anti-inflammatory herb, on esophageal cancer cells that produce interleukin 6. Cancer Lett 158:35–41

    Article  CAS  Google Scholar 

  12. Lee CH, Chen JC, Hsiang CY, Wu SL, Wu HC, Ho TY (2007) Berberine suppresses inflammatory agents-induced interleukin-1β and tumor necrosis factor-α productions via the inhibition of IκB-α degradation in human lung cells. Pharmacol Res 56:193–201

    Article  CAS  Google Scholar 

  13. Fukuda K, Hibiya Y, Mutoh M, Koshiji M, Asao S, Fujiwara H (1999) Inhibition by berberine of cyclooxygenase-2 transcriptional activity in human colon cancer cells. J Ethnopharmacol 66:227–233

    Article  CAS  Google Scholar 

  14. Jiang Q, Liu P, Wu X, Liu W, Shen X, Lan T, Xu S, Peng J, Xie X, Huang H (2011) Berberine attenuates lipopolysaccharide-induced extracellular matrix accumulation and inflammation in rats masangial cells: involvement of NF-κB signaling pathway. Mol Cell Endocrinol 331:34–40

    Article  CAS  Google Scholar 

  15. Kim HY, Koh EJ, Park J, Lee SM (2010) Gardenia jasminoides prevents galactosamine-induced acute hepatitis in rats. Yakhak Hoechi 54:403–409

    CAS  Google Scholar 

  16. Pan T, Shi X, Chen H, Chen R, Wu D, Lin Z, Zhang J, Pan J (2017) Geniposide suppresses interleukin-1β-induced inflammation and apoptosis in rat chondrocytes via the PI3K/Akt/NF-κB signaling pathway. Inflammation. https://doi.org/10.1007/s10753-017-0694-2

    Article  PubMed  Google Scholar 

  17. Xu GL, Li G, Ma HP, Zhong H, Liu F, Ao GZ (2009) Preventive effect of crocin in inflamed animals and in LPS-challenged RAW264.7 cell. J Agric Food Chem 57:8325–8330

    Article  CAS  Google Scholar 

  18. Yang K, Min Z, Shi Y, Xiang L, Meng Y, Wu C, Huang W, Tang B (2009) Effects of total iridoid glycosides from Fructus Gardeniae on inflammatory reaction and neuronal apoptosis in rats with intracerebral hemorrhage. Zhongyao Xinyao Yu Linchuang Yaoli 20:8–10

    Google Scholar 

  19. Lim H, Park KR, Lee DU, Kim YS, Kim HP (2008) Effects of the constituents of Gardenia Fructus on prostaglandin and NO production. Biomol Therapeut 16:82–86

    Article  CAS  Google Scholar 

  20. Koo HJ, Lim KH, Jung HJ, Park EH (2006) Anti-inflammatory evaluation of gardenia extract, geniposide and genipin. J Ethnopharmacol 103:496–500

    Article  CAS  Google Scholar 

  21. Koo HJ, Song YS, Kim HJ, Lee YH, Hong SM, Kim SJ, Kim BC, Jun C, Lim CJ, Park EH (2004) Antiinflammatory effects of genipin, an active principle of gardenia. J Pharmacol 495:201–208

    CAS  Google Scholar 

  22. Zeng H, Dou S, Zhao J, Fan S, Yuan X, Zhu S, Li L, Zhong W, Liu R (2011) The inhibitory activities of the components of Huang-Lian-Jie-Du-Tang (HLJDT) on eicosanoid generation via lipoxygenase pathway. J Ethnopharmacol 135:561–568

    Article  CAS  Google Scholar 

  23. Lu J, Wang JS, Kong LY (2011) Anti-inflammatory effects of Huang-Lian-Jie-Du decoction, its two fractions and four typical components. J Ethnopharmacol 134:911–918

    Article  CAS  Google Scholar 

  24. Oshima N, Narukawa Y, Hada N, Kiuchi F (2013) Quantitative analysis of anti-inflammatory activity of orengedokuto: importance of combination of flavonoids in inhibition of PGE2 production in mouse macrophage-like cell line J774.1. J Nat Med 67:281–288

    Article  CAS  Google Scholar 

  25. Kim KW, Ha KT, Park CS, Jin UH, Chang HW, Lee IS, Kim CH (2007) Polygonum cuspidatum, compared with baicalin and berberine, inhibits inducible nitric oxide synthase and cyclooxygenase-2 gene expressions in RAW-264.7 macrophage. Vascul Pharmacol 47:99–107

    Article  CAS  Google Scholar 

  26. Fujii T, Okuyama T, Wakame K, Okumura T, Ikeya Y, Nishizawa M (2017) Identification of anti-inflammatory constituents in Phellodendri Cortex and Coptidis Rhizoma by monitoring the suppression of nitric oxide production. J Nat Med 71:745–756

    Article  CAS  Google Scholar 

  27. Zhou K, Hu L, Liao W, Yin D, Rui F (2016) Coptisine prevent IL-β-induced expression of inflammatory mediators in chondrocytes. Inflammation 39:1558–1564

    Article  CAS  Google Scholar 

  28. Wu J, Zhang H, Hu B, Yang L, Wang P, Wang F, Meng X (2016) Coptisine from Coptis chinensis inhibits production of inflammatory mediators in lipopolysaccharide-induced RAW 264.7 murine macrophage cells. Eur J Pharmacol 780:106–114

    Article  CAS  Google Scholar 

  29. Wang JR, Tanaka T, Zhang H, Kouno I, Jiang ZH (2012) Formation and conformation of baicalin–berberine and wogonoside–berberine complexes. Chem Pharm Bull 60:706–711

    Article  CAS  Google Scholar 

  30. Yi L, Xu X (2004) Study on the precipitation reaction between baicalin and berberine by HPLC. J Chromatogr B 810:165–168

    Article  CAS  Google Scholar 

  31. Huang L, Fuchino H, Kawahara N, Narukawa Y, Hada N, Kiuchi F (2016) Application of a new method, orthogonal projection to latent structure (OPLS) combined with principal component analysis (PCA), to screening of prostaglandin E2 production inhibitory flavonoids in Scutellaria Root. J Nat Med 70:731–739

    Article  CAS  Google Scholar 

  32. Shimizu T, Shibuya N, Narukawa Y, Oshima N, Hada N, Kiuchi F (2018) Synergistic effect of baicalein, wogonin and oroxylin A mixture: multistep inhibition of the NF-κB signalling pathway contributes to an anti-inflammatory effect of Scutellaria Root flavonoids. J Nat Med 72:181–191

    Article  CAS  Google Scholar 

  33. The Ministry of Health, Labour and Welfare of Japan (2016) The Japanese pharmacopoeia. 17th edn. The Ministry of Health, Labour and Welfare of Japan, Tokyo, p 1747

  34. Dirsch VM, Stuppner H, Vollmar AM (1998) The griess assay: suitable for a bio-guided fractionation of anti-inflammatory plants extract. Planta Med 64:423–426

    Article  CAS  Google Scholar 

  35. Park KH, Park M, Choi SE, Jeong MS, Kwon JH, Oh MH, Choi HK, Seo SJ, Lee MW (2009) The Anti-oxidative and anti-inflammatory effects of caffeoyl derivatives from the roots of Aconitum Koreanum R. RAYMOND. Biol Pharm Bull 32:2029–2033

    Article  CAS  Google Scholar 

  36. Rubinstein LV, Shoemaker RH, Paull KD, Simon RM, Tosini S, Skehan P, Scudiero DA, Monks A, Boyd MR (1990) Comparison of in vitro anticancer-drug-screening data generated with a tetrazolium assay versus a protein assay against a diverse panel of human tumor cell lines. J Natl Cancer Inst 82:1113–1118

    Article  CAS  Google Scholar 

  37. Wu S, Sun A, Liu R (2005) Separation and purification of baicalin and wogonoside from the Chinese medicinal plant Scutellaria baicalensis Geroge by high-speed counter-current chromatography. J Chromatogr A 1066:243–247

    Article  CAS  Google Scholar 

  38. Abe K, Inoue O, Yumioka E (1990) Biliary excretion of metabolites of baicalin and baicalein in rats. Chem Pharm Bull 38:208–211

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported in part by MEXT-Supported Program for the Strategic Research Foundation at Private Universities from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Authors and Affiliations

  1. Faculty of Pharmacy, Keio University, 1-5-30 Shibakoen, Minato-ku, Tokyo, 105-8512, Japan

    Naohiro Oshima, Tomofumi Shimizu, Yuji Narukawa, Noriyasu Hada & Fumiyuki Kiuchi

  2. Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510, Japan

    Naohiro Oshima & Noriyasu Hada

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  1. Naohiro Oshima
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  2. Tomofumi Shimizu
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Correspondence to Fumiyuki Kiuchi.

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Oshima, N., Shimizu, T., Narukawa, Y. et al. Quantitative analysis of the anti-inflammatory activity of orengedokuto II: berberine is responsible for the inhibition of NO production. J Nat Med 72, 706–714 (2018). https://doi.org/10.1007/s11418-018-1209-7

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  • DOI: https://doi.org/10.1007/s11418-018-1209-7

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