DOI QR코드

DOI QR Code

현삼(玄蔘) 추출물이 RBL-2H3 비만세포에서 β-hexosaminidase 및 cytokine 분비에 미치는 효과

Inhibitory Effects of Scrophulariae Radix on β-hexosaminidase release and cytokine production in RBL-2H3 cells

  • 김세기 (대구가톨릭대학교 제약산업공학과)
  • Kim, Se-Gie (Department of Pharmaceutical Science and Technology)
  • 투고 : 2017.10.10
  • 심사 : 2017.11.15
  • 발행 : 2017.11.30

초록

Objectives : Traditional medicines isolated from natural products often have positive effects in the prevention and healing of various immune disorders, such as allergy and atopic inflammation. Scrophulariae Radix (SR) been used in oriental medicine used for treatment of acute and chronic inflammatory diseases. Mast cells are known to play important roles in the initiation of allergic reactions. In this study, we investigated the effects of SR ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : Rat basophilic leukemia RBL-2H3 cells were purchased from Korean Cell Line Bank (KCLB No. 22256). Cell viability was measured by MTT assay. Assays for ${\beta}-Hexosaminidase$ Secretion : RBL-2H3 cells were sensitized with dinitrophenyl-ImmunoglobulinE (DNP IgE). The next antigen DNP-BSA ($25ng/m{\ell}$) was added for 10 minutes and the reaction was terminated after 5 minutes in the ice bath. To determine ${\beta}-Hexosaminidase$ release, supernatants were aliquoted into 96-well plates. Samples were mixed with substrate solution and incubated for 1 h at $37^{\circ}C$. Absorbance was measured with a spectrophotometer at 405 nm. IL-4 and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) concentrations in cell culture supernatants were measured using enzyme-linked immunosorbent assay (ELISA) kits. Results : The cytotoxicity of SRE in RBL-2H3 cells was less than 5%. SRE inhibited DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. Also significantly decreased the levels of inflammatory cytokine, IL-4 and TNF-alpha. In this study, the SRE showed potential anti-allergic and antiinflammatory. Conclusions : These results indicate that SRE could be inhibit the allergic response through suppressing the mast cell activation.

키워드

참고문헌

  1. Baroody FM, Naclerio RM. Antiallergic effects of H1- receptor antagonists. Allergy . 2000 ; 55(64):17-27.
  2. Moon PD, Na HJ and Kim HM. Action of enzyme food, Green Life Enzyme of systemic and local ana-phylaxis. J OPEM. 2003 ; 3(1) : 46-50.
  3. Sin TY. allergy and Oriental medicine, Sin-il, seoul 2010 ; p. 13.
  4. Schwartz LB, Lewis RA, Seldin D, Austen KF. Acid hydrolases and tryptase from secretory granules of dispersed human lung mast cells. J Immunol . 1981 ;126(4) : 1290-4.
  5. Marquardt DL, Wasserman SI. Modulation of rat serosal mast cell biochemistry by in vivo dexamethasone administration. J Immunol . 1983 ; 131(2) : 934-9.
  6. Mastuda H, Morikawa T, Ueda K, Managi H, Yoshikawa M. Structural requirements of flavonoids for inhibition of antigen-Induced degranulation, TNF-${\alpha}$ and IL-4 production from RBL-2H3 cells. Bioorg Med Chem. 2002 ; 10(10) : 3123-8. https://doi.org/10.1016/S0968-0896(02)00227-4
  7. Lee MY, Seo CS, Lee JA, Lee NH, Kim JH, Ha HK, Zheng MS, Son JK, Shin HK. Anti-asthmatic effects of Angelica dahurica against ovalbumin-induced airway inflammation via upregulation of heme oxygenase-1. Food Chem Toxicol. 2011 ; 49(4) ; 829-837. https://doi.org/10.1016/j.fct.2010.12.004
  8. Vo TS, Kong CS, Kim SK. Inhibitory effects of chitooligosaccharides on degranulation and cyt kine generation in rat basophilic leukemia RBL-2H3 cells. Carbohyd Polym 2011 ; 84(1) : 649-655. https://doi.org/10.1016/j.carbpol.2010.12.046
  9. Lee SH, Bae EA, Park EK, Shin YW, Baek NI, Han EJ, Chung HG, Kim DH. Inhibitory effect of eupatilin and jaceosidin isolated from Artemisia princepsin IgE-induced hypersensitivity. Int Immunopharmacol. 2007 ; 7(13) : 1678-1684. https://doi.org/10.1016/j.intimp.2007.08.028
  10. Choi YH, Jung JW. Anxiolytic-like Effects of Scrophularia buergeriana Miquel Using the Elevated Plus-Maze in Mice - Involvement of GABAergic Nervous System. Korean J Oriental Physiology & Pathology . 2010 ; 24(3) : 476-483.
  11. Yook KD. Antimicrobial activity and cytotoxicity test of Scrophularia ningpoensis hemsl extracts against Klebsiella pneumoniae. Journal of The Korea Society of Computer and Information . 2016 ; 21(5) : 135-139. https://doi.org/10.9708/JKSCI.2016.21.5.135
  12. Park KH, Ham IH, Choi HY. Hypotensive Effect of Scrophularia buergeriana and Gene Analysis of Scrophularia Species. J of Herbology . 2008 ; 23(2) : 213-23.
  13. Song JS, Lee JC, Choi JH, Kim JH, Park SY. Effects of Scrophulariae Radix (SR) on Allergic Contact Dermatitis (ACD) induced by DNCB in mice. The Journal of Korean Oriental Medical Ophthalmology & Otolaryngology & Dermatology . 2011 ; 24(3) : 1-16.
  14. Yeo EJ, HAN JK, Kim YH. Inhibitory Studies of Scrophulariae Radix on Development of Dermatitis in NC/Nga Mice. J Korean Oriental Pediatrics. 2007 ; 21(2) : 13-34.
  15. Park CU, Cho CS, Kim CJ. The Effects of Radix Scrophulariae on the Hyperthyroidism of Rats. J Korean Oriental Med . 2006 ; 27(3) : 178-186.
  16. Park YK, Kang BS. Effects of Scrophulariae radix on Antioxidation in Rat's live. J of Herbology. 1998 ; 13(1) : 201-20.
  17. Kim SR, Kang SY, Lee KY, Kim SH, Markelonis GJ, Oh TH, Kim YC. Anti-amnestic activity of E-Pmethoxycinnamic acid from Scrophularia buergeriana. Cognitive Brain Research. 2003 ; 17(2) : 454-61. https://doi.org/10.1016/S0926-6410(03)00161-7
  18. Ahmed B, Al-Rehaily AJ, A1-Howiriny TA, El- Sayed KA, Ahmad MS. Scropolioside-D2 and harpagoside-B : two new iridoid glycosides and antiinflammatory activity. Biol. Pharm. Bull . 2003 ; 26(4) : 462-7. https://doi.org/10.1248/bpb.26.462
  19. Lee EJ, Kim SR, Kim J, Kim YC. Hepatoprotective Phenylpropanoids from Scrophularia buergeriana roots against CCl4-Induced toxicity: Action mechanism and structure-activity relationship. Planta Med . 2002 ; 68(5) : 407-11. https://doi.org/10.1055/s-2002-32081
  20. Tohda C, Kakihara Y, Komatsu K, Kuraishi, Y. lnhibitory effects of methanol extracts of herbal medicines on substance P-induced itch-scratch response. Biol. Pharm. Bull. 2000 ; 23(5) : 599-601. https://doi.org/10.1248/bpb.23.599
  21. Iikura M, Yamaguchi M, Hirai K, Miyamasu M, Yamada H, Nakajima T, et al. Regulationof surface FcepsilonRI expression on human eosinophils by IL-4 and IgE. International Achives of Allergy & Immunology. 2001 ; 124(4) : 470-7. https://doi.org/10.1159/000053782
  22. Govan A, Macfarlane P, Callander R, Lee JD. Pathology illustrated. 2nd ed. Seoul : Koryeohwahak. 1991 : 29.
  23. Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB. Evaluation of a tetrazolium based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res . 1987 ; 47(4) : 936-942.
  24. Park YJ. Allergy drug developments. KISTI. Seoul: KISTI. 2006: 1-5.
  25. Brown JM, Wilson TM, Metcalfe DD. The mast cell and allergic diseases: role in pathogenesis and implications for therapy. Clin. Exp. Allergy 2008;38:4-18.
  26. Hu ZQ, Zhao WH, Shimamura T. Regulation of mast cell development by inflammatory factors. Curr Med Chem. 2007;14(28):3044-50. https://doi.org/10.2174/092986707782793998
  27. Gordon JR. Burd PR. Galli SJ. Mast cells as a source multifunctional cytokines. Immunol. Today. 1990;11:458-64. https://doi.org/10.1016/0167-5699(90)90176-A
  28. Song Z, Casolaro V, Chen R. Georas S.N, Monos D, Ono, S.J, Polymorphic nucleotides within the human IL-4 promoter that mediate overexpression of the gene. J Immunol 1996 ;156(2) : 424-429.
  29. Egle P, Neil F. The RBL-2H3 cell line: its provenance and suitability as a model for the mast cell. Inflamm Res. 2009 ; 58(11) : 737-45. https://doi.org/10.1007/s00011-009-0074-y
  30. Kim D, Kim SH, Park EJ, Kang CY, Cho SH, Kim S. Anti-allergic effects of PG102, a water-soluble extract prepared from Actinidia arguta, in a murine ovalbumin-induced asthma model Clin Exp Allergy. 2009;39(2):280-9. https://doi.org/10.1111/j.1365-2222.2008.03124.x
  31. Ahn K. Role of mast cells in allergic inflammation and innate immunity. Kor J Pediatrics . 2004;47(11):1137-1141.