1. INFLAMMATION IN ALZHEIMER’S DISEASES AND THE RECOVER BY CANNABIONOIDS
Diana Aguirre-Rueda , Patricio Paredes-Brunet and Soraya L. Valles
Department of Physiology, School of Medicine, University of Valencia, Spain.
Introduction
Many studies have shown the relationship between inflammation and Alzheimer’s disease (AD). The hallmark in AD is both accumulation of beta-
amyloid (Aβ) plates and the presence of TAU protein inside neurons. Furthermore, glial cell activation, occurs after plates appear in brain damaged,
producing astrogliosis and microglia activation. Our group has shown inflammation in astrocytes in primary culture comparing Aβ with control cells.
Here we determined the action of cannabinoids in Aβ inflammation in astrocytes in culture. Protein expression levels were detected by western-blot and
ELISA techniques in astrocytes in primary culture treated with Aβ and/or cannabinoids. Using Aβ (10 μM) during 24 h, an increase of pro-inflammatory
mediators (NFκB, TNF-α and IL-1β), compared with control astrocytes was detected. Treatment with Win 55, 212-2 (10 μM) produced increase of anti-
inflammatory mediators (PPAR-γ) and decrease of pro-inflammatory mediators, such as NFκB, TNF-α and IL-1β, protecting cells to the toxic action of
Aβ. In transgenic mice (APP/Preseniline I), using microarray to principal inflammatory proteins, we detect induction of pro-inflammatory mediators and
reduction of anti-inflammatory mediators compared with wild type mice. With those results we demonstrate an unbalance between inflammatory and
anti-inflammatory mediators in Alzheimer’s disease and a possible positive effect of cannabinoids in AD.
Fig. 1. Infalmmatory cytoquine IL-1β levels in astrocytes in primary culture. Fig. 2. Infalmmatory cytoquine TNF-α levels in astrocytes in primary culture.
IL-1β levels were determined by ELISA in astrocytes treated for 24 h with TNF-α levels were determined by ELISA in astrocytes treated for 24 h with 10
10 μM Aβ (40-1) (C), 10 μM Aβ (1-42) (Aβ), 10 μM WIN 55, 212-2 (WIN) μM Aβ (40-1) (C), 10 μM Aβ (1-42) (Aβ), 10 μM WIN 55, 212-2 (WIN) and 10 μM
and 10 μM Aβ (1-42) + 10 μM WIN 55, 212-2 (Aβ + WIN). Data are means Aβ (1-42) + 10 μM WIN 55, 212-2 (Aβ + WIN). Data are means ± D.S. * p < 0.05
± D.S. * p < 0.05 vs control cells # p < 0,05 vs Aβ treated cells. n = 3. vs control cells # p < 0,05 vs Aβ treated cells. n = 3.
Fig. 3.- Inflamatory mediators NFκB expression in astrocytes in primary Fig. 4.- Anti-inflamatory mediators PPAR-γ expression in astrocytes in
culture. NFκB expression were determined by western-blot in astrocytes primary culture. PPAR-γ expression were determined by western-blot in
treated for 24 h with 10 μM Aβ (40-1) (C), 10 μM Aβ (1-42) (Aβ), 10 μM astrocytes treated for 24 h with 10 μM Aβ (40-1) (C), 10 μM Aβ (1-42) (Aβ),
WIN 55, 212-2 (WIN) and 10 μM Aβ (1-42) + 10 μM WIN 55, 212-2 (Aβ + 10 μM WIN 55, 212-2 (WIN) and 10 μM Aβ (1-42) + 10 μM WIN 55, 212-2
WIN). Data are means ± D.S. * p < 0.05 vs control cells # p < 0,05 vs Aβ (Aβ + WIN). Data are means ± D.S. * p < 0.05 vs control cells # p < 0,05 vs
treated cells. n = 3. Aβ treated cells. n = 3.
Fig. 5.- Microarray of pro and anti-inflammatory proteins, transgenic (T) vs. wild type (C). RNA obtained from 7th months cortex samples of female wild type
and transgenic mice. Microarray determined different pro-inflammatory and anti-inflammatory mediators.
Fig. 6.- Microarray of pro and anti-inflammatory proteins, wild type (C) vs. transgenic (T). RNA obtained from 7th months cortex samples of female wild type and
transgenic mice. Microarray determined different pro-inflammatory and anti-inflammatory mediators.
CONCLUSION
Our findings suggest that aβ in able to induce inflammatory mediator in transgenic mice (APP/PS1), also
in astrocytes, and these effect are prevented by the use of cannabinoids.