Fig 1: Effects of successive PGJ2 microinfusions on the following factors of the PGD2/J2 prostaglandin pathway: COX-2 (a), L-PGDS (b), DP2 receptor (c), and 15-PGDH (d), all in dopaminergic neurons and microglia, and 15-PGDH also in oligodendrocytes (e). Immunostaining for COX-2 (red in a), L-PGDS (red in b), DP2 (red in c), 15-PGDH (red in d and e), TH+ neurons (blue in a–d), Iba1+ microglia (green in a–d), and GST-pi+ oligodendrocytes (green in e) at 4 weeks after two (2X) and four (4X) PGJ2 microinjections. Scale bar = 50 μm. a COX-2 is significantly increased in dopaminergic neurons from rats receiving four (4X) PGJ2 injections compared to controls. Co-localization of COX-2 and Iba1 is greater in microglia from rats receiving two (2X) PGJ2 injections than in controls. b L-PGDS is significantly increased in dopaminergic neurons from rats receiving four (4X) PGJ2 injections than in controls. Co-localization of L-PGDS and Iba1 is greater in microglia from rats receiving two (2X) and four (4X) PGJ2 injections than in controls. c DP2 levels remain stable in dopaminergic neurons from all treatment groups, but DP2+ staining is almost absent in microglia. d 15-PGDH expression is increased in dopaminergic neurons from rats receiving four (4X) PGJ2 injections, but not in microglia. e 15-PGDH is highly expressed in SNpc oligodendrocytes from all groups of rats. Values on the y-axis represent the optical density (OD, normalized to TH, left graphs), or co-localization (normalized to Iba1, right graphs) ratios between ipsilateral SNpc over the contralateral. Black circles, control, DMSO-treated rats; red circles, PGJ2-treated rats. Statistical significance was estimated with Student’s T test to compare DMSO and PGJ2-treated groups. The p value in red indicates significant (p < 0.05) difference from DMSO-injected rats. N = 3 rats per group. X = number of injections (once per week)
Fig 2: Scheme depicting novel and previously tested therapeutic agents (in green) that could have potential to treat PD. (1) DP2 receptor antagonists: in the SNpc, DP2 receptors were detected in dopaminergic neurons but not in microglia. DP2 receptor activation leads to a decrease in cAMP and an increase in calcium. These effects mediate neurotoxicity induced by DP2 receptor activation. Thus, DP2 receptor antagonists could potentially prevent DA neurodegeneration. (2) COX-inhibitors: Ibuprofen, a COX inhibitor, prevented many of the sequelae induced by the PGJ2 treatment. Our data support the epidemiological studies that report the beneficial effect of NSAIDs in lowering the risk of developing PD. (3) L-PGDS inhibitors: in the brain, this enzyme synthesizes PGD2 from PGH2. PGD2 and its metabolite PGJ2 can be neurotoxic. Thus, decreasing L-PGDS activity will lower PGD2 levels and could prevent or diminish the neurotoxic effects of PGD2/J2 on dopaminergic neurons. COX 1 and 2, cyclooxygenase 1 and 2; L-PGDS, lipocalin prostaglandin D synthase; 15-PGDH, 15-hydroxyprostaglandin dehydrogenase; PPARγ, peroxisome proliferator-activated receptor gamma; PET, positron emission tomography (see text for details)
Fig 3: Brain infiltration of group 2 innate lymphoid cells (ILC2s) following cerebral ischaemia. (A,B) ILC2s were counted in the brain tissue of healthy subjects (controls) and patients who had an ischaemic stroke during the early phase (<24 hours). Images and bar graph show brain-infiltrating CRTH2+CD127+ cells in brain sections obtained from patients who died of acute ischaemic stroke. Scale bar=50 µm (insert=20 µm). n=7 subjects (4 men and 3 women) in the stroke group; n=9 subjects (5 men and 4 women) in the control group. **p<0.01 according to Mann-Whitney test. (C,D) Male and female C57BL/6 (B6) mice were subjected to sham operation or middle cerebral artery occlusion (MCAO). The presence of ILC2s was evaluated in the brains of sham control or MCAO mice. Images and bar graph show brain-infiltrating CD3e−ST2+ ILC2s from sham or MCAO mice. Scale bar=50 µm (insert=20 µm). n=11 mice per group. *p<0.05 according to two-tailed unpaired Student’s t-test. (E) Flow cytometry plots show mouse ILC2s (Lin−CD45highCD90.2+ ST2+, Lin=CD3e, CD45R, CD11b, Ter119, Ly-6G, CD11c, NK1.1, CD4, CD5, CD8a, TCR-β and TCR-γδ) in the brain, peripheral blood, lung and spleen at day 1 after MCAO. (F,G) Summarised results show ILC2s numbers in the brain, peripheral blood, lung and spleen from sham or MCAO mice at day 1 and day 3 after MCAO. n=9 mice per group. *p<0.05, **p<0.01 according to Mann-Whitney test. Data are presented as the mean±SD.
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