I. Paterniti, M. Campolo, M. Cordaro, D. Impellizzeri, R. Siracusa, R. Crupi, E. Esposito, and S. Cuzzocrea. University of Messina, Italy
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1 I. Paterniti, M. Campolo, M. Cordaro, D. Impellizzeri, R. Siracusa, R. Crupi, E. Esposito, and S. Cuzzocrea University of Messina, Italy
2 A spinal cord lesion is a partial or total disruption of the structural or functional integrity of the spinal cord, often caused by some trauma, resulting in impairments such as paralysis, loss of sensation and autonomic nervous system dysfunction. The most common causes of SCI are automobile crashes (31.5%) and falls (25.3%), followed by gunshot wounds (10.4%), motorcycle crashes (6.8%), diving incidents (4.7%), and medical/surgical complications
3 Nowadays, SCI affects a significant number of patients worldwide.
4
5
6 Properties of Melatonin
7
8 PPARs
9 The aim of the present study was to better understand the mechanism of action of melatonin in reducing the inflammatory process and the oxidative stress associated to spinal cord injury (SCI). Moreover, we hypotized that melatonin exert its action binding on nuclear receptors, such as PPARα receptor. Thus we perform an in vivo model of spinal cord injury both in WT mice and in PPARα KO mice.
10 Methods and Parameters 1h 6h 12h 24h 10 days SCI Sacrifice Histology H&E WB Immunohistochemistry Immunofluorescence Sacrifice BMS score
11 Histological score PPARαKO The severity of tissue damage following SCI is decreased in Melatonin WT treated mice but no in PPARαKO mice Sham SCI SCI-Mel Sham SCI SCI-Mel 5 4 *** 3 2 ### 1 ND 0 Sham WT WT SCI+Melatonin WT ND Sham PPAR a KO SCI PPAR a KO CI+Melatonin PPAR a KO
12 MPO Levels (U/mg protein) Effect of Melatonin Neutrophil infiltration (MPO assay) *** 20 ### 10 0 Sham WT WT SCI+Melatonin WT Sham PPAR a KO PPAR a KO SCI+Melatonin PPAR a KO
13 Densitometric Units (OD*mm 2 ) Densitometric Units (OD*mm 2 ) Effect of Melatonin on inflammatory pathways IκBα degradation and NFκB translocation CD 1 Ppar α KO Sham SCI SCI + Mel Sham SCI SCI + Mel CD 1 Ppar α KO Sham SCI SCI + Mel Sham SCI SCI + Mel IκBα 7kDa NFκBp65 65kDa *** *** ### ## 0 Sham CD1 CD1 SCI+Melatonin CD1 Sham PPAR a KO PPAR a KO SCI+Melatonin PPAR a KO 0 Sham CD1 CD1 SCI+Melatonin CD1 Sham PPAR a KO PPAR a KO SCI+Melatonin PPAR a KO
14 TNF-a (pg/100 mg wet tissue) IL-1b (pg/100 mg wet tissue) Effect of Melatonin on pro-inflammatory cytokines production 120 *** *** ### ### 20 0 Sham WT WT SCI+Melatonin WT Sham PPAR a KO PPAR a KO SCI+Melatonin PPAR a KO 0 Sham WT WT SCI+Melatonin WT Sham PPAR a KO PPAR a KO SCI+Melatonin PPAR a KO
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17 Densitometric Units (OD*mm 2 ) Densitometric Units (OD*mm 2 ) Densitometric Units (OD*mm 2 ) Densitometric Units (OD*mm 2 ) Effect of Melatonin on MAPK pathway CD 1 Ppar α KO p-p38 Sham SCI SCI + Mel Sham SCI SCI + Mel p-akt p-erk p-jnk * ### p-akt Β-actin 0 Sham CD1 CD1 SCI+Melatonin CD1 Sham PPAR a KO PPAR a KO SCI+Melatonin PPAR a KO ** p-p ** p-erk ** p-jnk ## 5000 ### 0 Sham CD1 CD1 SCI+Melatonin CD1 Sham PPAR a KO PPAR a KO SCI+Melatonin PPAR a KO 0 Sham CD1 CD1 SCI+Melatonin CD1 Sham PPAR a KO PPAR a KO SCI+Melatonin PPAR a KO 0 Sham CD1 CD1 SCI+Melatonin CD1 Sham PPAR a KO PPAR a KO SCI+Melatonin PPAR a KO
18 Effect of Melatonin on locomotor recovery at 10 days
19 Take home message The results obtained indicate that the anti-inflammatory activity of melatonin is weakened in PPAR-α KO mice, compared to WT controls as evaluated by inhibition of the degree of tissue injury, neutrophil infiltration, pro-inflammatory cytokine expressions, NF-κB activation and MAPK expression. This study indicates that PPAR- can contribute to the anti-inflammatory activity of melatonin in SCI.
20 . I would like to thank the following people who have contributed so generously to these results Prof. Salvatore Cuzzocrea Prof.ssa Emanuela Esposito Michela Campolo Marika Cordaro Rosalba Siracusa Daniela Impellizzeri Rosalia Crupi Giuseppe Bruschetta
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