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Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (2): 145-151.doi: 10.3736/jcim20100209

• Original Experimental Research • Previous Articles     Next Articles

Effects of astilbin on maturation and immunologic function of mouse bone marrow-derived dendritic cells

 Shao-hua SONGa, Xiao-yun SHENb, Guo-shan DINGa, Fang LIUa, Zhen-meng WANGc, Zhi-ren FUa   

  1. a Organ Transplantation Center, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
    b Institute of Immunology, Second Military Medical University, Shanghai 200433, China
    c Department of Anesthesiology, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China
  • Received:2009-11-05 Accepted:2009-12-30 Online:2010-02-20 Published:2010-02-15
  • Contact: Zhi-ren FU E-mail:zhirenfu@163.com

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Objective

To explore the effects of astilbin on the maturation and immunologic function of mouse bone marrow-derived dendritic cells (DCs).
Methods

Mouse bone marrow cells were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor and interleukin-4 (IL-4) for 5 days to get immature DCs (imDCs), then the imDCs was cultured in the presence of 1 μg/mL lipopolysaccharide (LPS) or LPS (1 μg/mL) plus astilbin (25, 50, 100 μg/mL) for 48 h. Then, the cells were harvested, and the apoptosis, immunophenotypes and antigen phagocytosis capability of imDCs in LPS, and low-, medium- and high-dose astilbin groups were analyzed by flow cytometry. Contents of p40 subunit of interleukin-12 (IL-12p40) in the supernatants were detected with enzyme-linked immunosorbent assay (ELISA). The stimulatory activity of the harvested cells on allogeneic T cells in mixed lymphocyte reactions (MLR) was tested by incorporation of 3H-thymidine, and the contents of IL-2, IL-4, IL-10 and interferon-γ (INF-γ) in the supernatants of MLR were examined by ELISA.
Results

At the concentrations of 25 to 100 μg/mL, astilbin exhibited no toxicity on co-cultured DCs. Compared with the lipopolysaccharide, low-, medium- and high-dose astilbin could decrease the expression levels of major histocompatibility complex-Ⅰa (MHC-Ⅰa), CD40, CD80 and CD86 molecules in DCs. DCs in the low-, medium- and high-dose astilbin groups exhibited weaker capabilities for antigen phagocytosis and less contents of IL-12p40 in the supernatants than in the LPS group. Furthermore, low-, medium- and high-dose astilbin showed weak activities in stimulating the proliferation of allogeneic T cells as compared with the LPS (P<0.05). Compared with the LPS, low-, medium- and high-dose astilbin could decrease IL-2 and INF-γ secretion from T cells in MLR but had no effect on IL-10 secretion.
Conclusion

Astilbin can inhibit maturation of mouse bone marrow-derived DCs with dose-dependent effect and exert negative effects on immunologic function of the DCs.

Key words: Astilbin, Dendritic cells, Immunologic function, Mice

Figure 1

Effects of astilbin on apoptosis of bone marrow dendritic cells"

Table 1

Expressions of MHC-Ⅰa and co-stimulatory molecules in DCs in different groups ($\bar{x}±s$, %)"

Group n MHC-Ⅰa CD40 CD80 CD86
imDC 4 38.24±5.68 22.89±3.18 26.79±3.55 36.22±5.17
Lipopolysaccharide 4 84.78±9.28** 60.69±6.16** 75.08±8.65** 60.33±7.25**
Low-dose astilbin 4 66.37±7.03△ 48.89±5.78△ 55.25±6.18△ 50.25±4.28△
Medium-dose astilbin 4 60.59±5.38△ 45.38±3.97△ 50.72±5.19△ 47.43±3.89△
High-dose astilbin 4 58.06±5.09△ 33.98±4.87△ 46.14±3.57△ 45.04±3.22△

Figure 2

Effects of astilbin on antigen phagocytosis of DCs The phagocytic ability of DCs was tested by measuring FITC-dextran phagocytosis by using fluorescence-activated cell sorting. Right histograms represent DCs cultured with 25 μg/mL FITC-dextran at 37 ℃ for 2 h and are marked with geometric mean fluorescence intensity. Left histograms represent cells cultured with 25 μg/mL FITC-dextran at 4 ℃ for 2 h as a control. A: imDC group; B: Lipopolysaccharide group; C: Low-dose astilbin group; D: Medium-dose astilbin group; E: High-dose astilbin group."

Figure 3

Effects of astilbin on secretion of IL-12p40 from DCs A: imDC group; B: Lipopolysaccharide group; C: Low-dose astilbin group; D: Medium-dose astilbin group; E: High-dose astilbin group. Data were represented as x±s, n=4. **P<0.01, vs imDC group; △P<0.05, vs lipopolysaccharide group."

"

Group n Stimulatory cells︰responsive cells
DCs︰T cells =1︰5 DCs︰T cells =1︰10 DCs︰T cells =1︰20
imDC 4 26.25±2.35 20.16±2.29 18.90±1.15
Lipopolysaccharide 4 58.66±10.61** 50.22±9.26** 45.78±7.68**
Low-dose astilbin 4 40.15±6.24△ 38.57±5.24△ 35.68±4.48△
Medium-dose astilbin 4 37.27±4.29△ 35.68±4.57△ 31.27±4.09△
High-dose astilbin 4 36.02±5.99△ 33.98±4.87△ 30.148±3.98△

Figure 4

Effects of astilbin on cytokine secretion from T cells in MLR when the proportion of DCs (Ratio of DCs to T cells) was 1︰10A: imDC group; B: Lipopolysaccharide group; C: Low-dose astilbin group; D: Medium-dose astilbin group; E: High-dose astilbin group. Data were represented as x±s, n=4. **P<0.01, vs imDC group; △P<0.05, vs lipopolysaccharide group."

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