In:
Neuroimmunomodulation, S. Karger AG, Vol. 23, No. 2 ( 2016), p. 109-121
Abstract:
〈 b 〉 〈 i 〉 Objective: 〈 /i 〉 〈 /b 〉 The aim of this study was to examine whether the natural protease inhibitor Av-cystatin (rAv17) of the parasitic nematode 〈 i 〉 Acanthocheilonema viteae 〈 /i 〉 exerts anti-inflammatory effects in an in vitro model of lipopolysaccharide (LPS)-activated microglia. 〈 b 〉 〈 i 〉 Methods: 〈 /i 〉 〈 /b 〉 Primary microglia were harvested from the brains of 2-day-old Wistar rats and cultured with or without rAv17 (250 n 〈 smlcap 〉 M 〈 /smlcap 〉 ). After 6 and 24 h the release of nitric oxide (Griess reagent) and TNF-α (ELISA) was measured in the supernatant. Real-time PCR was performed after 2, 6 and 24 h of culture to measure the mRNA expression of IL-1β, IL-6, TNF-α, COX-2, iNOS and IL-10. To address the involved signaling pathways, nuclear NF- & #x0138;B translocation was visualized by immunocytochemistry. Morphological changes of microglia were analyzed by Coomassie blue staining. Differences between groups were calculated using one-way ANOVA with Bonferroni's post hoc test. 〈 b 〉 〈 i 〉 Results: 〈 /i 〉 〈 /b 〉 Morphological analysis indicated that LPS-induced microglial transformation towards an amoeboid morphology is inhibited by rAv17. Av-cystatin caused a time-dependent downregulation of proinflammatory cytokines, iNOS and COX-2 mRNA expression, respectively. This was paralleled by an upregulated expression of IL-10 in resting as well as in LPS-stimulated microglia. Av-cystatin reduced the release of NO and TNF-α in the culture supernatant. Immunocytochemical staining demonstrated an attenuated translocation of NF- & #x0138;B by Av-cystatin in response to LPS. In addition, Western blot analysis revealed a rAv17-dependent reduction of the LPS-induced ERK1/2-pathway activation. 〈 b 〉 〈 i 〉 Conclusion: 〈 /i 〉 〈 /b 〉 The parasite-derived secretion product Av-cystatin inhibits proinflammatory mechanisms of LPS-induced microglia with IL-10, a potential key mediator.
Type of Medium:
Online Resource
ISSN:
1021-7401
,
1423-0216
Language:
English
Publisher:
S. Karger AG
Publication Date:
2016
detail.hit.zdb_id:
1483035-8
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