Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
권호 표지
DOI QR코드

DOI QR Code

Isolation and quantitative analysis of metabolites from Scrophularia buergeriana and their hepatoprotective effects against HepG2 Cells

현삼 (Scrophularia buergeriana)에서 분리한 화합물의 함량분석 및 간세포 보호 효과

  • Na, Hyeon Seon (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Oh, Seon Min (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Shin, Woo Cheol (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Bo, Jeon Hwang (Graduate School of Biotechnology and Department of Oriental Medicine Biotechnology, Kyung Hee University) ;
  • Kim, Hyoung-Geun (Graduate School of Biotechnology and Department of Oriental Medicine Biotechnology, Kyung Hee University) ;
  • Yoon, Dahye (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Yang, Seung Hwan (Department of Biotechnology, Chonnam National University) ;
  • Lee, Young-Seob (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Kim, Geum-Soog (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA) ;
  • Baek, Nam-In (Graduate School of Biotechnology and Department of Oriental Medicine Biotechnology, Kyung Hee University) ;
  • Lee, Moon-Soon (Department of Industrial Plant Science and Technology, Chungbuk National University) ;
  • Lee, Dae Young (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA)
  • Received : 2019.10.14
  • Accepted : 2019.11.04
  • Published : 2019.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

The roots of Scrophularia buergeriana were extracted with 80% aqueous Methanol and the concentrates were partitioned into EtOAc, n-BuOH, and H2O fractions. The repeated silica gel or octadecyl SiO2column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of phenylethanoid glycosides and iridoid glycosides. The chemical structures of these compounds were determined as harpagoside (1), angoroside C (2), aucubin (3) and acetoside (4) based on spectroscopic analyses including nuclear magnetic resonance and MS. A simple and efficient HPLC with UV detection method for the simultaneous determination of the four compounds (1-4) has been developed and applied to their content determination in the S. buergeriana. The roots were extracted by 80% methanol, and the contents of 1, 2, 3, and 4 were determined to 11.5, 7.6, 41.2, and 4.8 mg/g, respectively. Additionally, angoroside C (2) and acetoside (4) exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

현삼(Scrophularia buergeriana) 뿌리를 80% Methanol수용액으로 추출한 뒤, 감압 농축한 추출물을 EtOAc, n-BuOH과 H2O층으로 계통 분획을 실시하였다. n-BuOH분획에 대하여 silica gel, octadecyl SiO2 column chromatograph 및 중압분취(MPLC) 장비를 반복 실시하여 4종의 phenylethanoid glycoside 및 iridoid glycoside계의 화합물을 분리하였다. NMR 및 Mass데이터를 해석하여, harpagoside (1), angoroside C (2), aucubin (3) 및 acetoside (4)로 구조 동정하였다. 분리한 4종의 화합물에 대하여 HPLC 분석법을 이용하여 정량분석한 결과, 11.5 mg/g (1), 7.6 mg/g (2), 41.2 mg/g (3), 및 4.8 mg/g (4) 이 현삼 뿌리에 함유된 것을 확인하였다. 현삼으로부터 분리된 화합물 중 angoroside C 및 acetoside는 에탄올에 의해 저해된 세포 성장률을 검증한 결과, 간암세포종인 HepG2세포에 대해서 간세포를 보호하는 효과가 있음을 확인하였다.

Keywords

References

  1. Lee JH, Park SM, Kim YC, Woo HJ (2001) A clinical study of the effects of Chungganhaeju-tang on alcoholic fatty liver. J Korean Med 22(4): 107-113
  2. Baik TH (2016) A case study on a patient with non-alcoholic fatty liver disease (NAFLD) by using ultrasound. J Korean Med 37(1): 151-157 https://doi.org/10.13048/jkm.16014
  3. Kwon TD (2007) The effects of exercise on hepatotoxicity in mice with alcoholic fatty liver. Gene Expr 11(3): 257-264
  4. Park SU, Li X, Eom SH, Lee CY, Lee SY (2010) E-p-methoxycinnamic acid production in hairy root cultures of Scrophularia buergeriana Miquel. Arch Biol Sci 62: 649-652 https://doi.org/10.2298/ABS1003649P
  5. Herbal Medicine School Compilation Committee (2013) Pharmacognosy. Dongmyeong Publisher, Gyeonggi-do, Korea, p 224
  6. Lee HW, Park SY, Choo BK, Chun JM, Lee AY, Kim HK (2007) Quantitative analysis comparison of korea and china Scrophulariae radix. Kor J Pharmacogn 38(1): 15-18
  7. Kim HM, Ahn MJ, SH Lee (2012) Isolation and identification of phytochemical constituents from Scrophularia takesimensis. J Med Plants Res 6(22): 3923-3930
  8. Kim SH, Kim DH, Park JH, Choi EJ, Park SH, Lee KY, Jeon MJ, Kim YC, Sung SH (2010) Discrimination of Scrophularia spp. according to geographic origin with HPLC-DAD combined with multivariate analysis. Microchem J 94(2): 118-124 https://doi.org/10.1016/j.microc.2009.09.013
  9. Lee MK, Choi OG, Park JH, Cho HJ, Ahn MJ, Kim SH, Kim YC, Sung SH (2007) Simultaneous determination of four active constituents in the roots of Scrophularia buergeriana by HPLC-DAD and LC-ESI-MS. J Sep Sci 30(15): 2345-2350 https://doi.org/10.1002/jssc.200700117
  10. Lee HJ, Spandidos DA, Tsatsakis A, Margina D, Izotov BN, Yang SH (2019) Neuroprotective effects of Scrophularia buergeriana extract against glutamate-induced toxicity in SH-SY5Y cells. Int J Mol Med 43(5): 2144-2152
  11. Jeong EJ, Ma CJ, Lee KY, Kim SH, Sung SH, Kim YC (2009) KD-501, a standardized extract of Scrophularia buergeriana has both cognitiveenhancing and antioxidant activities in mice given scopolamine. J Ethnopharmacol 121(1): 98-105 https://doi.org/10.1016/j.jep.2008.10.006
  12. Park KH, Ham IH, Choi HY (2008) Hypotensive effect of Scrophularia buergeriana and gene analysis of Scrophularia Species. Kor J Herbology 23(2): 213-223
  13. Kim SG (2017) Inhibitory effects of Scrophulariae radix on ${\beta}$-hexosaminidase release and cytokine production in RBL-2H3 cells. Kor J Herbology 32(6): 9-15 https://doi.org/10.6116/kjh.2017.32.6.9
  14. Byun SH, Yang CH, Kim SC (2005) Inhibitory effect of Scrophulariae Radix extract on$TNF-{\alpha}$, IL-$1{\beta}$, IL-6 and nitric oxide production in lipopolysaccharide-activated RAW 264.7 cell. Kor J Herbology 20(2): 7-16
  15. Shin H, Medriano CA, Park B, Park YH, Lee KY (2018) Screening and identification of neuroprotective compounds from Scrophularia buergeriana using cell extraction coupled with LC-MS. J Pharm Biomed Anal 148: 355-360 https://doi.org/10.1016/j.jpba.2017.10.018
  16. Lv PY, Feng H, Huang WH, Tian YY, Wang YQ, Qin YH, Li XH, Hu K, Zhou HH, Ouyang DS (2017) Aucubin and its hydrolytic derivative attenuate activation of hepatic stellate cells via modulation of $TGF-{\beta}$ stimulation. Environ Toxicol Pharmacol 50: 234-239 https://doi.org/10.1016/j.etap.2017.02.012
  17. Lin SJ, Jiang SH, Li YM, Zeng JF, Zhu DY (2000) Two novel iridoids from Scrophularia buergeriana. Tetrahedron Lett 41(7): 1069-1071 https://doi.org/10.1016/S0040-4039(99)02244-3
  18. Wu XM, Zhang LQ, Chen XC, Feng L, Xing WX, Li YM (2014) A new iridoid derivative from the roots of Scrophularia buergeriana. Acta Pharmacol Sin 49(7): 1019-1021
  19. Kim SR, Lee KY, Koo KA, Sung SH, Lee NG, Kim JW, Kim YC (2002) Four new neuroprotective iridoid glycosides from Scrophularia buergeriana Roots. J Nat Prod 65(11): 1696-1699 https://doi.org/10.1021/np0202172
  20. Jeong EJ, Lee KY, Kim SH, Sung SH, Kim YC (2008) Cognitiveenhancing and antioxidant activities of iridoid glycosides from Scrophularia buergeriana in scopolamine-treated mice. Eur J Pharmacol 588(1): 78-84 https://doi.org/10.1016/j.ejphar.2008.04.015
  21. Tundis R, Loizzo MR, Menichini F, Statti GA, Menichini F (2008) Biological and pharmacological activities of iridoids: recent developments. Mini Rev Med Chem 8(4): 399-420 https://doi.org/10.2174/138955708783955926
  22. Zhang L, Zhu T, Qian F, Xu J, Dorje G, Zhao Z, Guo FJ, Li Y (2014) Iridoid glycosides isolated from Scrophularia dentata Royle ex Benth. and their anti-inflammatory activity. Fitoterapia 98: 84-90 https://doi.org/10.1016/j.fitote.2014.07.005
  23. Kim SR, Kim YC (2000) Neuroprotective phenylpropanoid esters of rhamnose isolated from roots of Scrophularia buergeriana. Phytochem 54(5): 503-509 https://doi.org/10.1016/S0031-9422(00)00110-2
  24. Diaz AM, Abad MJ, Fernandez L, Silvan AM, De Santos J, Bermejo P (2004) Phenylpropanoid glycosides from Scrophularia scorodonia: In vitro anti-inflammatory activity. Life Sci 74(20): 2515-2526 https://doi.org/10.1016/j.lfs.2003.10.008
  25. Carvalho AA, Andrade LN, de Sousa EBV, de Sousa DP (2015) Antitumor phenylpropanoids found in essential oils. Biomed Res Int. doi: 10.1155/2015/392674
  26. Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, Kenney S, Boyd MR (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J Nat Cancer Inst 82(13): 1107-1112 https://doi.org/10.1093/jnci/82.13.1107
  27. Li YM, Jiang SH, Gao WY, Zhu DY (1999) Iridoid glycosides from Scrophularia ningpoensis. Phytochem 50(1): 101-104 https://doi.org/10.1016/S0031-9422(98)00477-4
  28. de Santos J, Lanza AMD, Fernandez L, Rumbero A (2000) Isoangoroside C, a phenylpropanoid glycoside from Scrophularia scorodonia roots. Z Naturforsch C 55(5-6): 333-336
  29. Ersoz T, Berkman MZ, Tasdemir D, Calis I, Ireland CM (2002) Iridoid and phenylethanoid glycosides from Euphrasia pectinata. Turk J Chem 26(2): 178-188
  30. Kim HG, Oh HJ, Ko JH, Song HS, Lee YG, Kang SC, Lee DY, Baek NI (2019) Lanceoleins A-G, hydroxychalcones, from the flowers of Coreopsis lanceolata and their chemopreventive effects against human colon cancer cells. Bioorg Chem 85: 274-281 https://doi.org/10.1016/j.bioorg.2019.01.003
  31. Andary C, Wylde R, Laffite C, Privat G, Winternitz F (1982) Structures of verbascoside and orobanchoside, caffeic acid sugar esters from Orobanche rapum-genistae. Phytochem 21(5): 1123-1127 https://doi.org/10.1016/S0031-9422(00)82429-2
  32. Mostafa M, Nahar N, Mosihuzzaman M, Makhmoor T, Choudhary MI, Rahman AU (2007) Free radical scavenging phenylethanoid glycosides from Leucas indica Linn. Nat Prod Res 21(4): 354-361 https://doi.org/10.1080/14786410701194401
  33. Li M, Xu T, Zhou F, Wang M, Song H, Xiao X, Lu B (2018) Neuroprotective effects of four phenylethanoid glycosides on $H_{2}O_{2}$-induced apoptosis on PC-12 cells via the Nrf2/ARE pathway. Int J Mol Sci 19(4): 1135 https://doi.org/10.3390/ijms19041135