Background: CHF5074, nonsteroidal anti-inflammatory derivative with g -secretase modulatory activity has been shown to inhibit brain plaque deposition and to attenuate or reverse memory deficit in different transgenic mouse models of Alzheimer disease (AD). The recent discovery of modulation Rho-GTPase-dependent signaling suggests that the range of biological actions exerted by this drug may be wider than expected. Since neuroinflammation is considered a major pathogenetic mechanism in AD, we investigated the possible regulation of microglia activation by CHF5074 in vivo and in vitro. Methods: The Tg2576 transgenic mouse carrying a transgene coding for the 695-amino acid isoform of human APP derived from a large Swedish family with early-onset AD was used for in vivo study. CHF5074 60 mg/kg was administered for 2 months to female transgenic and wild-type mice of 7 months of age (n ¼ 12/group). Activated microglia was measured by Iba1 immunohistochemistry, area fraction and cell number evaluation. In vitro experiments were carried out on microglia enriched rat primary cultures, activated by LPS 0.1mM for 8 h, and CHF5074 pretreatment was performed at 1, 3 and 10mM one hour before LPS administration. TNFalfa, IL-6, IL-12 and RANTES were measured by ELISA in the cell culture medium. Results: In vivo, quantification of immunoreactivity indicated that compared to transgenic controls, wild-type animals had much lower activated microglia (cell count and area fraction, P<0.05). Compared to Tg2576 control mice, activated microglia in the cerebral cortex was significantly (P < 0.05) reduced by CHF5074 treatment. In vitro experiments indicated that RANTES up-regulation by LPS was significantly attenuated by CHF5074 3 and 10 mM. Conclusions: This results of this study showed that CHF5074 significantly inhibits neuroinflammatory activity mediated by microglia. RANTES (regulated upon activation, normal T cell expressed and secreted) regulation could support a specific role in microglia-astrocyte cross-talk.
IN VIVO AND IN VITRO STUDY ON MICROGLIA ACTIVATION OF CHF5074, A NONSTEROIDAL ANTI-INFLAMMATORY DERIVATIVE WITH GAMMA-SECRETASE MODULATORYACTIVITY / Tomasini, Cristina; Fernandez, Mercedes; Sivilia, Sandra; Pietrini, Vladimiro; Imbimbo, Bruno; Calza, Laura; Antonelli, Tiziana; Giardino, Luciana. - In: ALZHEIMER'S & DEMENTIA. - ISSN 1552-5260. - 8:(2012), pp. 301-302. (Intervento presentato al convegno AAIC 2012 tenutosi a Vancouver, Canada nel 14-19 luglio 2012).
IN VIVO AND IN VITRO STUDY ON MICROGLIA ACTIVATION OF CHF5074, A NONSTEROIDAL ANTI-INFLAMMATORY DERIVATIVE WITH GAMMA-SECRETASE MODULATORYACTIVITY
PIETRINI, Vladimiro;
2012-01-01
Abstract
Background: CHF5074, nonsteroidal anti-inflammatory derivative with g -secretase modulatory activity has been shown to inhibit brain plaque deposition and to attenuate or reverse memory deficit in different transgenic mouse models of Alzheimer disease (AD). The recent discovery of modulation Rho-GTPase-dependent signaling suggests that the range of biological actions exerted by this drug may be wider than expected. Since neuroinflammation is considered a major pathogenetic mechanism in AD, we investigated the possible regulation of microglia activation by CHF5074 in vivo and in vitro. Methods: The Tg2576 transgenic mouse carrying a transgene coding for the 695-amino acid isoform of human APP derived from a large Swedish family with early-onset AD was used for in vivo study. CHF5074 60 mg/kg was administered for 2 months to female transgenic and wild-type mice of 7 months of age (n ¼ 12/group). Activated microglia was measured by Iba1 immunohistochemistry, area fraction and cell number evaluation. In vitro experiments were carried out on microglia enriched rat primary cultures, activated by LPS 0.1mM for 8 h, and CHF5074 pretreatment was performed at 1, 3 and 10mM one hour before LPS administration. TNFalfa, IL-6, IL-12 and RANTES were measured by ELISA in the cell culture medium. Results: In vivo, quantification of immunoreactivity indicated that compared to transgenic controls, wild-type animals had much lower activated microglia (cell count and area fraction, P<0.05). Compared to Tg2576 control mice, activated microglia in the cerebral cortex was significantly (P < 0.05) reduced by CHF5074 treatment. In vitro experiments indicated that RANTES up-regulation by LPS was significantly attenuated by CHF5074 3 and 10 mM. Conclusions: This results of this study showed that CHF5074 significantly inhibits neuroinflammatory activity mediated by microglia. RANTES (regulated upon activation, normal T cell expressed and secreted) regulation could support a specific role in microglia-astrocyte cross-talk.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
