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01.12.2017 | Research | Ausgabe 1/2017 Open Access

Journal of Neuroinflammation 1/2017

Polysaccharides from Ganoderma lucidum attenuate microglia-mediated neuroinflammation and modulate microglial phagocytosis and behavioural response

Zeitschrift:
Journal of Neuroinflammation > Ausgabe 1/2017
Autoren:
Qing Cai, Yuanyuan Li, Gang Pei
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​s12974-017-0839-0) contains supplementary material, which is available to authorized users.

Abstract

Background

Ganoderma lucidum (GL) has been widely used in Asian countries for hundreds of years to promote health and longevity. The pharmacological functions of which had been classified, including the activation of innate immune responses, suppression of tumour and modulation of cell proliferations. Effective fractions of Ganoderma lucidum polysaccharides (GLP) had already been reported to regulate the immune system. Nevertheless, the role of GLP in the microglia-mediated neuroinflammation has not been sufficiently elucidated. Further, GLP effect on microglial behavioural modulations in correlation with the inflammatory responses remains to be unravelled. The aim of this work was to quantitatively analyse the contributions of GLP on microglia.

Methods

The BV2 microglia and primary mouse microglia were stimulated by lipopolysaccharides (LPS) and amyloid beta42 (Aβ42) oligomer, respectively. Investigation on the effect of GLP was carried by quantitative determination of the microglial pro- and anti-inflammatory cytokine expressions and behavioural modulations including migration, morphology and phagocytosis. Analysis of microglial morphology and phagocytosis modulations was confirmed in the zebrafish brain.

Results

Quantitative results revealed that GLP down-regulates LPS- or Aβ-induced pro-inflammatory cytokines and promotes anti-inflammatory cytokine expressions in BV-2 and primary microglia. In addition, GLP attenuates inflammation-related microglial migration, morphological alterations and phagocytosis probabilities. We also showed that modulations of microglial behavioural responses were associated with MCP-1 and C1q expressions.

Conclusions

Overall, our study provides an insight into the GLP regulation of LPS- and Aβ-induced neuroinflammation and serves an implication that the neuroprotective function of GLP might be achieved through modulation of microglial inflammatory and behavioural responses.
Zusatzmaterial
Additional file 1: Figure S1. GLP exhibited no cytotoxicity and proliferative effect to BV2 cells. (A) GLP effect on BV2 cell viability was examined for 24 h in the presence and absence of LPS stimulation. No cytotoxicity was detected. GLP long-term effect on cell growth was investigated for 48, 72 and 96 h. Cell growth was estimated by total cell count (B) and CellTiter-Glo assay (C). Cells were incubated in normal DMEM culture medium with 10% FBS supplement. Both 1 and 0.01 μg/ml showed no effect on cell growth compared to untreated control. Cells grown in FBS-depleted medium (2% FBS) showed little cell growth, whereas in the FBS-enriched medium (30% FBS), the cell growth exceeds normal cultured medium. Figure S2. Confirmation on the integrity of prepared Aβ oligomer. Four microliters of oligomers or fibril were loaded to the nitrocellulose membrane. Dot blot was performed and the oligomer-(A11) and Fibril-specific (ab126468) antibodies were used. In the prepared oligomer (left lane), little fibril fractions were detected, whilst in the prepared Aβ fibril (right lane), certain levels of oligomers were also detected. Staining of 6E10 revealed the total Aβ content. (PDF 207 kb)
12974_2017_839_MOESM1_ESM.pdf
Literatur
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