Nat Prod Sci.  2017 Dec;23(4):239-246. 10.20307/nps.2017.23.4.239.

Inhibitory Effects of Methanol Extract from Nardostachys chinensis on 27-hydroxycholesterol-induced Differentiation of Monocytic Cells

Affiliations
  • 1Department of Pharmacology, School of Medicine, Pusan National University, Yangsan 50612, Republic of Korea. koanhoi@pusan.ac.kr
  • 2Division of Pharmacology, School of Korean Medicine, Pusan National University, Yangsan 50612, Republic of Korea.
  • 3Department of Microbiology and Immunology, Pusan National University - School of Medicine, Yangsan, Gyeongnam 50612, Republic of Korea.
  • 4Institute of Marine BioTechnology, Pusan National University, Busan, Republic of Korea.

Abstract

27-Hydroxycholesterol (27OHChol) has been reported to induce differentiation of monocytic cells into a mature dendritic cell phenotype. We examined the effect of methanol extract of Nardostachys chinensis (Nard) on 27OHChol-induced differentiation using THP-1, a human monocytic cell line. Treatment of monocytic cells with methanol extract of Nard resulted in decreased transcription and surface expression of CD80, CD83, and CD88 elevated by 27OHChol in a dose-dependent manner. Surface levels of MHC class I and II molecules elevated by 27OHChol were also reduced to basal levels by treatment with the Nard extract. Decreased endocytosis activity caused by 27OHChol was recovered by treatment with the Nard extract. CD197 expression and cell attachment were attenuated by the Nard extract. In addition, levels of transcription and surface expression of CD molecules involved in atherosclerosis, such as CD105, CD137, and CD166 upregulated by 27OHChol were significantly decreased by treatment with methanol extract of Nard. These results indicate that methanol extract of Nard down-regulates 27OHChol-induced differentiation of monocytic cells into a mature dendritic cell phenotype and expression of CD molecules associated with atherosclerosis. The current study suggests that biological activity of oxygenated cholesterol derivatives can be inhibited by herbal medication.

Keyword

Dendritic cells; Differentiation; 27-Hydroxycholesterol; Monocyte; Nardostachys chinensis

MeSH Terms

Atherosclerosis
Cell Line
Cholesterol
Dendritic Cells
Endocytosis
Humans
Methanol*
Monocytes
Nardostachys*
Oxygen
Phenotype
Cholesterol
Methanol
Oxygen

Figure

  • Fig. 1 Effects of Nard extract on the transcription and surface expression of mDC-markers induced by 27OHChol. (A) THP-1 cells (1 × 106 cells/60 mm culture dish) were cultured for 48 h with 2.5 µg/ml of 27OHChol with or without 5, 25 or 50 µg/ml of Nard extract. Transcription of CD80, CD83, and CD88 was analyzed by real-time PCR. Data are expressed as the mean ± SD (n = 3 replicates/group). (B) THP-1 cells (1 × 106 cells/60 mm culture dish) were cultured for 48 h in the presence of 2.5 µg/ml of 27OHChol with or without 5, 25 or 50 µg/ml of Nard extract. Cells were immunostained with antibodies against CD80, CD83, and CD88 and analyzed by flow cytometry. Results are representative of three independent experiments.

  • Fig. 2 Effects of Nard extract on expression of MHC class I and II molecules induced by 27OHChol. Following incubation for 48 h with 27OHChol (2.5 µg/ml) with or without indicated concentrations of Nard extract, the stimulated THP-1 cells (1 × 106 cells/60 mm culture dish) were immunostained for MHC class I and II. Fluorescence was analyzed by flow cytometry. Results are representative of three independent experiments.

  • Fig. 3 Effects of Nard extract on functional alteration of monocytic cells induced by 27OHChol. Following incubation for 48 h with 27OHChol (2.5 µg/ml) with or without indicated concentrations of Nard extract, THP-1 cells were treated with 1 mg/ml of FITC-conjugated dextran for 1 h. The cells were analyzed by flow cytometry. Results are representative of three independent experiments.

  • Fig. 4 Effects of Nard extract on cell adhesion and expression of CD197 induced by 27OHChol. (A) THP-1 cells (1 × 106 cells/60 mm culture dish) were cultured for 48 h with 2.5 µg/ml of 27OHChol with or without indicated concentrations of Nard extract. The harvested cells were immunostained with an anti-CD197 antibody and analyzed by flow cytometry. Results are representative of three independent experiments. (B) THP-1 cells (2 × 104 cells/well of 96-well plate) were cultured for 48 h with 2.5 µg/ml of 27OHChol with or without indicated concentrations of Nard extract. Suspended cells in the well were removed, and the adherent cells were counted. Data are expressed as the mean ± SD (n = 3 replicates/group).

  • Fig. 5 Effects of Nard extraction expression of atherosclerosis-associated CD molecules induced by 27OHChol. (A) Following incubation with 27OHChol (2.5 µg/ml) for 48 h with or without indicated concentrations of Nard extract, transcription of CD105, CD137, and CD166 was analyzed by real-time PCR. Data are expressed as the mean ± SD (n = 3 replicates/group). (B) THP-1 cells (1 × 106 cells/60 mm culture dish) were cultured for 48 h with 2.5 µg/ml of 27OHChol with or without indicated concentrations of Nard extract. The harvested cells were immunostained with antibodies against CD105, CD137, and CD166 and analyzed by flow cytometry. Results are representative of three independent experiments.


Reference

1. Sallusto F, Lanzavecchia A. J Exp Med. 1994; 179:1109–1118.
2. Zhou LJ, Tedder TF. Proc Natl Acad Sci U S A. 1996; 93:2588–2592.
3. Lyakh LA, Koski GK, Telford W, Gress RE, Cohen PA, Rice NR. J Immunol. 2000; 165:3647–3655.
4. Berges C, Naujokat C, Tinapp S, Wieczorek H, Höh A, Sadeghi M, Opelz G, Daniel V. Biochem Biophys Res Commun. 2005; 333:896–907.
5. Son Y, Kim SM, Lee SA, Eo SK, Kim K. Biochem Biophys Res Commun. 2013; 438:161–168.
6. Takaya Y, Takeuji Y, Akasaka M, Nakagawasai O, Tadano T, Kisara K, Kim K, Wataya Y, Niwa M, Oshima Y. Tetrahedron. 2000; 56:7673–7678.
7. Zhang J, Qiang CC, Li WJ, Liu LJ, Lin XX, Cheng YJ, Tang K, Yao FJ, Wu SH. J Cardiovasc Pharmacol. 2014; 64:127–133.
8. Takemoto H, Omameuda Y, Ito M, Fukuda T, Kaneko S, Akaike A, Kobayashi Y. Biol Pharm Bull. 2014; 37:1050–1055.
9. Hwang JS, Lee SA, Hong SS, Han XH, Lee C, Lee D, Lee CK, Hong JT, Kim Y, Lee MK, Hwang BY. Bioorg Med Chem Lett. 2012; 22:706–708.
10. Seo HC, Kim SM, Eo SK, Rhim BY, Kim K. Biomol Ther (Seoul). 2015; 23:84–89.
11. Steinman RM. Annu Rev Immunol. 1991; 9:271–296.
12. Banchereau J, Briere F, Caux C, Davoust J, Lebecque S, Liu YJ, Pulendran B, Palucka K. Annu Rev Immunol. 2000; 18:767–811.
13. Perry HM, McNamara CA. Arterioscler Thromb Vasc Biol. 2012; 32:1548–1549.
14. Profumo E, Buttari B, Saso L, Capoano R, Salvati B, Riganò R. Scientific World J. 2012; 2012:157534.
15. Katz FE, Parkar M, Stanley K, Murray LJ, Clark EA, Greaves MF. Eur J Immunol. 1985; 15:103–106.
16. Fujimoto Y, Tu L, Miller AS, Bock C, Fujimoto M, Doyle C, Steeber DA, Tedder TF. Cell. 2002; 108:755–767.
17. Gavett SH, O'Hearn DJ, Li X, Huang SK, Finkelman FD, Wills-Karp M. J Exp Med. 1995; 182:1527–1536.
18. Banchereau J, Steinman RM. Nature. 1998; 392:245–252.
19. Moschovakis GL, Bubke A, Dittrich-Breiholz O, Braun A, Prinz I, Kremmer E, Förster R. Eur J Immunol. 2012; 42:48–57.
20. DuSell CD, Umetani M, Shaul PW, Mangelsdorf DJ, McDonnell DP. Mol Endocrinol. 2008; 22:65–77.
21. Piao M, Tokunaga O. J Atheroscler Thromb. 2006; 13:82–89.
22. Olofsson PS, Söderström LA, Wågsäter D, Sheikine Y, Ocaya P, Lang F, Rabu C, Chen L, Rudling M, Aukrust P, Hedin U, Paulsson-Berne G, Sirsjö A, Hansson GK. Circulation. 2008; 117:1292–1301.
23. Kim SM, Kim BY, Lee SA, Eo SK, Yun Y, Kim CD, Kim K. Toxicol Appl Pharmacol. 2014; 274:462–470.
Full Text Links
  • NPS
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr