1. OXIDATIVE STRESS AND INFLAMMATION IN ALZHEIMER´S DISEASE
1D. Aguirre-Rueda, 2C.
García-Lucerga, Sanchez-Pérez y Valles. 1A. 1S.L
1Departamento de Fisiología, Universidad de Valencia, España.
2Departamento de Fisioterapia, Universidad de Valencia, España.
Introduction
 Many studies have shown the relationship between oxidative stress and Alzheimer’s disease (AD). The hallmark in AD is both accumulation of beta-amyloid 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 oxidative stress and inflammation in neurons and
astrocytes in primary culture. Using microarray and RT-PCR we mid gene expression of oxidative stress and inflammation genes. Also, by western-blot we determine protein expression of inflammation and
oxidative stress markers in transgenic (APP/Preseniline I) compared with wild type mice.
 Our results show an induction of pro-inflammatory mediators and reduction of anti-inflammatory mediators in transgenic mice compared with wild type. Also pro-oxidants proteins are increased and anti-oxidants
proteins are decreased in transgenic mice compared with wild type. Our results demonstrate an unbalance between oxidants and antioxidants proteins and also an unbalance between inflammatory and anti
inflammatory mediators in transgenic mice compared with wild type.
Function Gene identifier Gene name Fold change Direction
Abcf1 ATP- binding cassette sub-family F (GCN20) member 1 2,190899034 Up
Bcl6 B-cell leukemia/lymphoma 6 1,073880901 Down
Blr1 Burkit lymphoma receptor 1
C3 Complement component 3 2,44336433 Down
Casp1 Caspase 1 1,188429235 Down
Chemokine Genes Ccl1 Chemokine (C-C motif) ligand 1
Ccl11 Small chemokine (C-C motif) ligand 11 125,6433061 Down
Ccl12 Chemokine (C-C motif) ligand 12 19,96916357 Down
Ccl17 Chemokine (C-C motif) ligand 17 31,44884221 Down
Ccl19 Chemokine (C-C motif) ligand 19 4,63108745 Down
Ccl2 Chemokine (C-C motif) ligand 2 1,199760966 Down
Ccl20 Chemokine (C-C motif) ligand 20
Ccl22 Chemokine (C-C motif) ligand 22 1,188159073 Down
Ccl24 Chemokine (C-C motif) ligand 24 1,949822776 Down
Ccl25 Chemokine (C-C motif) ligand 25 420,1381549 Down
Ccl3 Chemokine (C-C motif) ligand 3 3,370891052 Up
Ccl4 Chemokine (C-C motif) ligand 4 3,104198899 Up
Ccl5 Chemokine (C-C motif) ligand 5 1,572800843 Down
Ccl6 Chemokine (C-C motif) ligand 6 1,352626009 Down
Ccl7 Chemokine (C-C motif) ligand 7 12,22641045 Down
Ccl8 Chemokine (C-C motif) ligand 8 13,6617747 Down Fig. 2.- Microarray of pro and anti-inflammatory proteins, transgenic (T) vs. wild type (C). RNA
Ccl9 Chemokine (C-C motif) ligand 9 3,258265123 Down obtained from 7th months cortex samples of female wild type and transgenic mice. Microarray
Chemokine ReceptorsCcr1 Chemokine (C-C motif) receptor 1 3,147601716 Down determined different pro-inflammatory and anti-inflammatory mediators.
Ccr2 Chemokine (C-C motif) receptor 2 15,91510754 Down
Ccr3 Chemokine (C-C motif) receptor 3 451,3538177 Down
Ccr4 Chemokine (C-C motif) receptor 4 12,99372345 Down
Ccr5 Chemokine (C-C motif) receptor 5 16,58895381 Down
Ccr6 Chemokine (C-C motif) receptor 6 10,2553086 Down
Ccr7 Chemokine (C-C motif) receptor 7 4,804342487 Down
Ccr8 Chemokine (C-C motif) receptor 8
Ccr9 Chemokine (C-C motif) receptor 9 99,3292009 Down
Crp C-reactive protein , pentraxin-related
Chemochine Genes Cx3cl1 Chemokine (C-X3-C motif) ligand 1 1,722094127 Down
Cxcl1 Chemokine (C-X-C motif) ligand 1
Cxcl10 Chemokine (C-X-C motif) ligand 10 7,17520972 Down
Cxcl11 Chemokine (C-X-C motif) ligand 11 5,252946897 Down
Cxcl12 Chemokine (C-X-C motif) ligand 12 1,297114647 Up
Cxcl13 Chemokine (C-X-C motif) ligand 13 1,543493064 Down
Cxcl15 Chemokine (C-X-C motif) ligand 15
Cxcl4 Chemokine (C-X-C motif) ligand 4 10,87836626 Down
Cxcl5 Chemokine (C-X-C motif) ligand 5 3,647019719 Down
Cxcl9 Chemokine (C-X-C motif) ligand 9 30,3151068 Down
Chemochine Receptors xcr3
C Chemokine (C-X-C motif) receptor 3
Ccr10 Chemokine (C-X-C motif) receptor 10 26,05351281 Down
Cytokine Genes Ifng Interferon gamma
Il10 Interleukin 10
Cytokine Receptors Il10ra Interleukin receptor, alpha 1,626417902 Down
Fig. 3.- Microarray of pro and anti-inflammatory proteins, wild type (C) vs. transgenic (T). RNA obtained
Il10rb Interleukin receptor, beta 2,428618699 Down from 7th months cortex samples of female wild type and transgenic mice. Microarray determined different
Cytokine Genes Il11 Interleukin 11 1,021726457 Down pro-inflammatory and anti-inflammatory mediators.
Il13 Interleukin 13
Cytokine Receptors Il13ra1 Interleukin 13 receptor, alpha 1 1,39337347 Down
Cytokine Genes Il15 Interleukin 15 1,373355383 Down
Il16 Interleukin 16 7,437635259 Down 0,18
Wild type P53
Il17b Interleukin 17B 14,38089939 Down
0,16
Relative Densitometric Units
Il18 Interleukin 18 1,856765987 Down Transgenic
0,14
Il1a Interleukin 1 alpha 3,289380724 Down
Il1b Interleukin 1 beta 2,297379193 Down 0,12
Il1f6 Interleukin 1 family, member 6 0,10 *
Il1f8 Interleukin 1 family, member 8 0,08
*
Il1r1 Interleukin 1 receptor, type I 1,005220501 Down 0,06 *
il1r2 Interleukin 1 receptor, type II 1,110263413 Down
*
0,04
Il20 Interleukin 20
0,02
Cytokine Receptors Il2rb Interleukin 2 receptor, beta chain 1,241338864 Down
0,00
Il2rg Interleukin 2 receptor, gamma chain 2,381319172 Down P53
Cortex Hipocamp Limbic Cerebellum
Cytokine Genes Il3 Interleukin 3 37,08474645 Up
Il4 Interleukin 4 3,367385754 Down
Wild type
Relative Densitometric Units
Cytokine Receptors Il5ra Interleukin 5 receptor, alpha 2,579592178 Down 0,60
HIF
Transgenic
Il6ra Interleukin 6 receptor, alpha 1,783400838 Down
0,50 HIF
Il6st Interleukin 6 signal transducer 1,439890398 Down
Il8rb Interleukin 8 receptor, beta 0,40
Cytokine Genes Itgam Integrin alpha M 1,000635124 Up
PPAR-γ
Itgb2 Integrin beta 2 2,014762802 Down 0,30
Lta Lymphotoxin A *
0,20
Ltb Lymphotoxin B 1,873029991 Down *
Mif Macrophage megration inhibitory factor 1,50753657 Down 0,10
Tubuline
Scye1 Small inducible cytokine subfamily E, member 1 2,60739291 Down
Spp1 Secreted phosphoprotein 1 12,42332116 Down 0,00
Tgfb1 Transforming growth factor, beta 1 2,89172027 Down Cortex Hipocamp Limbic Cerebellum
4,5 C H L Ce / C H L Ce
Tnf Tumor necrosis factor
PPAR-γ wild type transgenic
Tnfrsf1a Tumor necrosis factor receptor superfamily, member 1a 1,904976235 Down 4
Wild Type
Relative Densitometric Units
Tnfrsf1b Tumor necrosis factor receptor superfamily, member 1b 1,047576063 Down 3,5 Transgenic
Cd4Olg CD40 ligand 3
Tollip Toll interacting protein 2,123415944 Down 2,5
Xcr1 Chemokine (C motif) receptor 1
2
Gusb Glucuronidase, beta 2,889892846 Down
1,5
Hprt1 Hypoxantine guanine phosphoribosyl transferase 1 2,051438636 Down
1
Hsp90ab1 Heat shock protein 90 kDa alpha (cytosolic) class B 1 2,2017032 Down
Gapdh Glyceraldehyde-3-phosphate dehydrogenase 1 Down 0,5
Actb Actin, beta, cytoplasmic 1,736553671 Down 0
MGDC Mouse genomic DNA Contamination cortex hipocamp limbic cerebellum
Fig. 4.- Western-blot from 7th months cortex samples of female wild type and transgenic mice. Western-blot
Fig. 1.- Microarray from cortex samples, wild type vs transgenic (APP/Presenilin 1) mice. RNA determined different pro-inflammatory and anti-inflammatory mediators.
obtained from 7th months cortex samples of female wild type and transgenic mice. Microarray
determined different pro-inflammatory and anti-inflammatory mediators.
CONCLUSIONS
1. Microarray determined an induction of pro-inflammatory and reduction of anti-inflammatory mediators in transgenic APP/Presenilin 1 compared with wild type.
2. Pro-inflammatory proteins are increased and anti-inflammatory proteins are decreased in transgenic compared with wild type, such as Fig 1, 2 and 3 shown.
3. Fig. 4 shows induction of PPAR-γ in hippocampus, limbic system and cerebellum from 7th months of female transgenic compared wild type. HIF protein are decreased in transgenic compared willd type and also p-
53 shows a decreases values in transgenic mice compared with wild type.
4. Our study demonstrated and unbalance between inflammatory and anti-inflammatory mediators in transgenic APP/Presenilin 1 mice compared with wild type.