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Volume 24, Issue 2, Pages 51-62 (June 2017) Silymarin attenuates aspartame-induced variation in mouse behaviour, cerebrocortical morphology and oxidative stress markers Adejoke Yetunde Onaolapo, Saratu Zayid Abdusalam, Olakunle James Onaolapo Pathophysiology Volume 24, Issue 2, Pages 51-62 (June 2017) DOI: 10.1016/j.pathophys.2017.01.002 Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 1 Effect of silymarin/aspartame co-administration on horizontal locomotion (upper panel) and rearing (lower panel) in mice. Each bar represents mean±SE.M. Comparisons are: *p<0.05 significantly different from VEH, #p<0.05 significantly different from silymarin, μp<0.05 aspartame alone significantly different from silymarin/aspartame co-administration. VEH: Vehicle, SILY: Silymarin, 160: aspartame at 160mg/kg, 320: aspartame at 320mg/kg, 160+SILY: aspartame at 160mg/kg co-administered with silymarin, 320+SILY: aspartame at 320mg/kg co-administered with silymarin, number of animals per group-10. Pathophysiology 2017 24, 51-62DOI: (10.1016/j.pathophys.2017.01.002) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 2 Effect of silymarin/aspartame co-administration on grooming behaviour in mice. Each bar represents mean±SE.M. Comparisons are: *p<0.05 significantly different from VEH, #p<0.05 significantly different from silymarin, μp<0.05 aspartame alone significantly different from silymarin/aspartame co-administration. VEH: Vehicle, SILY: Silymarin, 160: aspartame at 160mg/kg, 320: aspartame at 320mg/kg, 160+SILY: aspartame at 160mg/kg co-administered with silymarin, 320+SILY: aspartame at 320mg/kg co-administered with silymarin, number of animals per group-10. Pathophysiology 2017 24, 51-62DOI: (10.1016/j.pathophys.2017.01.002) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 3 Effect of silymarin/aspartame co-administration on spatial working-memory (upper panel) and Y-maze locomotor activity (lower panel) in mice. Each bar represents mean±SE.M. Comparisons are: *p<0.05 significantly different from VEH, #p<0.05 significantly different from silymarin, μp<0.05 aspartame alone significantly different from silymarin/aspartame co-administration. VEH: Vehicle, SILY: Silymarin, 160: aspartame at 160mg/kg, 320: aspartame at 320mg/kg, 160+SILY: aspartame at 160mg/kg co-administered with silymarin, 320+SILY: aspartame at 320mg/kg co-administered with silymarin, number of animals per group-10. Pathophysiology 2017 24, 51-62DOI: (10.1016/j.pathophys.2017.01.002) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 4 Effect of silymarin/aspartame co-administration on anxiety-related behaviour in mice. Each bar represents mean±SE.M. Comparisons are: *p<0.05 significantly different from VEH, #p<0.05 significantly different from silymarin, μp<0.05 aspartame alone significantly different from silymarin/aspartame co-administration. VEH: Vehicle, SILY: Silymarin, 160: aspartame at 160mg/kg, 320: aspartame at 320mg/kg, 160+SILY: aspartame at 160mg/kg co-administered with silymarin, 320+SILY: aspartame at 320mg/kg co-administered with silymarin, number of animals per group-10. Pathophysiology 2017 24, 51-62DOI: (10.1016/j.pathophys.2017.01.002) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 5 (a–f) Effect of silymarin/aspartame co-administration on cerebral cortex morphology in mice. 5a) Vehicle 5b) SILY 5c) ASP 160mg/kg, 5d) ASP 320mg/kg, 5e) ASP 160mg/kg+SILY, 5f) ASP 320mg/kg+SILY. Representative photomicrographs of hematoxylin and eosin (H&E) stained sections of the mouse cerebral cortex showing granule cell (GC), pyramidal cell (PC), neuroglia (NG), degenerating pyramidal cell (De PC). SILY: Silymarin, ASP 160: aspartame at 160mg/kg, ASP 320: aspartame at 320mg/kg, ASP 160+SILY: aspartame at 160mg/kg co-administered with silymarin, ASP 320+SILY: aspartame at 320mg/kg co-administered with silymarin, number of animals per group-5. H&E x160, scale bar: 29μm/pixel. Pathophysiology 2017 24, 51-62DOI: (10.1016/j.pathophys.2017.01.002) Copyright © 2017 Elsevier B.V. Terms and Conditions

Fig. 6 (a–f) Effect of silymarin/aspartame co-administration on GFAP reactivity of the cerebral cortex in mice. 6a) Vehicle 6b) SILY 6c) ASP 160mg/kg, 6d) ASP 320mg/kg, 6e) ASP 160mg/kg+SILY, 6f) ASP 320mg/kg+ SILY. Representative photomicrographs of glial fibrillary acid protein (GFAP) immunohistochemistry sections of the mouse cerebral cortex showing GFAP immunoreactive astrocytes (Gr As), pyramidal cell (PC), granule cells (GC). SILY: Silymarin, ASP 160: aspartame at 160mg/kg, ASP 320: aspartame at 320mg/kg, ASP 160+SILY: aspartame at 160mg/kg co-administered with silymarin, ASP 320+SILY: aspartame at 320mg/kg co-administered with silymarin, number of animals per group-5.GFAP IHC x100, Scale bar: 29μm/pixel. Pathophysiology 2017 24, 51-62DOI: (10.1016/j.pathophys.2017.01.002) Copyright © 2017 Elsevier B.V. Terms and Conditions