Acta Pharmacologica Sinica (2010) 31: 191–201; doi: 10.1038/aps.2009.205

Acta Pharmacologica Sinica (2010) 31: 191–201; doi: 10.1038/aps.2009.205

Original Article

Immunomodulatory activity of andrographolide on macrophage activation and specific antibody response complex

Wei WANG^1,2,#, Jing WANG^2,#, Sheng-fu DONG², Chun-hong LIU², Paola ITALIANI³, Shu-hui SUN², Jing XU⁴, Diana BORASCHI³, Shi-ping MA^1,*,Di QU^2,*

¹Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing 210038, China; ²Key Laboratory of Medical Molecular Virology of Ministries of Education and Health, Institute of Medical Microbiology and Institutes of Biomedical Sciences, Shanghai Medical College of Fudan University, Shanghai 200032, China; ³Laboratory of Cytokines, Unit of Immunobiology, Institute of Biomedical Technologies, National Research Council, Pisa 56124, Italy; ⁴Beijing Institute of Biological Products, Beijing 100024, China

Aim: To investigate the immunomodulatory effects of andrographolide on both innate and adaptive immune responses.

Methods: Andrographolide (10 µg/mL in vitro or 1 mg/kg in vivo) was used to modulate LPS-induced classical activated (M1) or IL-4-induced alternative activated (M2) macrophages in vitro and humor immune response to HBsAg in vivo. Cytokine gene expression profile (M1 vs M2) was measured by real-time PCR, IL-12/IL-10 level was detected by ELISA, and surface antigen expression was evaluated by flow cytometry, whereas phosphorylation level of ERK 1/2 and AKT was determined by Western blot. The level of anti-HBs antibodies in HBsAg immunized mice was detected by ELISA, and the number of HBsAg specific IL-4-producing splenocyte was enumerated by ELISPOT.

Results: Andrographolide treatment in vitro attenuated either LPS or IL-4 induced macrophage activation, inhibited both M1 and M2 cytokines expression and decreased IL-12/IL-10 ratio (the ratio of M1/M2 polarization). Andrographolide down-regulated the expression of mannose receptor (CD206) in IL-4 induced macrophages and major histocompability complex/costimulatory molecules (MHC I, CD40, CD80, CD86) in LPS-induced macrophages. Correspondingly, anti-HBs antibody production and the number of IL-4-producing splenocytes were reduced by in vivoadministration of andrographolide. Reduced phosphorylation levels of ERK1/2 and AKT were observed in macrophages treated with andrographolide.

Conclusion: Andrographolide can modulate the innate and adaptive immune responses by regulating macrophage phenotypic polarization and Ag-specific antibody production. MAPK and PI3K signaling pathways may participate in the mechanisms of andrographolide regulating macrophage activation and polarization.

Keywords: macrophages; cytokines; antibodies; immunomodulator; andrographolide

This work was supported by the Program of Ministry of Science and Technology of China (S2010GR0385, 2008ZX10004-014, and 2008ZX10002-002), the International Science and Technology Cooperation Program of Shanghai Science and Technology Commission (074307038) and PhD student visiting scholarship of Fudan University for 2008-2009.

^#These two authors contributed equally to this work.
* To whom correspondence should be addressed.
E-mail dqu@shmu.edu.cn (Di QU); spma@cpu.edu.cn (Shi-ping MA)
Received 2009-06-29 Accepted 2009-12-24

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