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

• Son Y
• Kim H
• Yang B
• Kim B
• Park YC
• Kim K

• KMID: 2401574
• DOI: http://doi.org/10.20307/nps.2017.23.4.239

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.

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