Insulin-like growth factor 1 prevents neuronal cell death and paraplegia in the rabbit model of spinal cord ischemia Yoshihisa Nakao, MD, Hajime Otani,

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Insulin-like growth factor 1 prevents neuronal cell death and paraplegia in the rabbit model of spinal cord ischemia Yoshihisa Nakao, MD, Hajime Otani, MD, Tadashi Yamamura, MD, Reiji Hattori, MD, Motohiko Osako, MD, Hiroji Imamura, MD The Journal of Thoracic and Cardiovascular Surgery Volume 122, Issue 1, Pages 136-143 (July 2001) DOI: 10.1067/mtc.2001.114101 Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 1 Blood glucose levels in the sham-operated (n = 5, open bars ), control (n = 8, shaded bars ), IGF-1-treated (n = 8, hatched bars ), and insulin-treated (n = 6, filled bars ) rabbits. *P <.01 com-pared with control rabbits. Ao, Aortic. The Journal of Thoracic and Cardiovascular Surgery 2001 122, 136-143DOI: (10.1067/mtc.2001.114101) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 2 Hematoxylin and eosin staining of the spinal cord segment (L6) 48 hours after spinal cord ischemia (upper panel ; original magnification 400×). The number of motor neuron cells in the motor anterior horn per section (lower panel ) is shown in the sham-operated group (n = 5), the control group (n = 8), the IGF-1-treated group (n = 8), and the insulin-treated group (n = 6). *P <.0001 compared with the control group. The Journal of Thoracic and Cardiovascular Surgery 2001 122, 136-143DOI: (10.1067/mtc.2001.114101) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 3 TUNEL staining of the spinal cord segment (L6) 48 hours after spinal cord ischemia (upper panel ; original magnification 200×). Arrowhead, TUNEL-negative motor neuron cells; arrow, TUNEL-positive motor neuron cells. Percentage of TUNEL-positive motor neuron cells in the motor anterior horn (lower panel ) is shown in the sham-operated group (n = 5), the control group (n = 8), the IGF-1-treated group (n = 8), and the insulin-treated group (n = 6). *P <.01 compared with the control group. †P <.001 compared with the IGF-1 group. The Journal of Thoracic and Cardiovascular Surgery 2001 122, 136-143DOI: (10.1067/mtc.2001.114101) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 4 Immunohistochemical staining for Bcl-xL (green fluorescence ) and Bax (red fluorescence ) in the spinal cord segment (L6) 24 and 48 hours after spinal cord ischemia. The fluorescence staining was visualized with confocal laser microscopy at a magnification of 600×. No appreciable expression of Bcl-xL and Bax was observed in motor neuron cells in the sham-operated rabbits. Expression of only Bax was increased in motor neuron cells in the control rabbits 24 and 48 hours after spinal cord ischemia. Treatment with IGF-1 increased expression of Bcl-xL in motor neuron cells, with no appreciable expression of Bax 24 and 48 hours after spinal cord ischemia. Treatment with insulin modestly increased Bcl-xL and inhibited Bax expression 24 hours after spinal cord ischemia, but only Bax expression was elevated 48 hours after the operation. The Journal of Thoracic and Cardiovascular Surgery 2001 122, 136-143DOI: (10.1067/mtc.2001.114101) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions

Fig. 5 Proposed mechanism of IGF-I-induced spinal cord protection. (+) and (-) indicate stimulation and inhibition, respectively. Dashed line indicates assault on mitochondria. NMDA, N- methyl-D -aspartate; NHE, Na+/H+exchange; Mt, mitochondria; cy.c, cytochrome c; AIF, apoptosis-inducing factor. The Journal of Thoracic and Cardiovascular Surgery 2001 122, 136-143DOI: (10.1067/mtc.2001.114101) Copyright © 2001 American Association for Thoracic Surgery Terms and Conditions