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Ethyl pyruvate modulates delayed paralysis following thoracic aortic ischemia reperfusion in mice  Bao-Ngoc Nguyen, MD, Hassan Albadawi, MD, Rahmi Oklu,

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Presentation on theme: "Ethyl pyruvate modulates delayed paralysis following thoracic aortic ischemia reperfusion in mice  Bao-Ngoc Nguyen, MD, Hassan Albadawi, MD, Rahmi Oklu,"— Presentation transcript:

1 Ethyl pyruvate modulates delayed paralysis following thoracic aortic ischemia reperfusion in mice 
Bao-Ngoc Nguyen, MD, Hassan Albadawi, MD, Rahmi Oklu, MD, PhD, Robert S. Crawford, MD, Mitchell P. Fink, MD, Richard P. Cambria, MD, Michael T. Watkins, MD  Journal of Vascular Surgery  Volume 64, Issue 5, Pages (November 2016) DOI: /j.jvs Copyright © Terms and Conditions

2 Fig 1 Temporal schematic diagram of the experimental protocol. Mice were treated with 300 mg/kg ethyl pyruvate (EP) 30 minutes before clamping the thoracic aorta and the left subclavian artery for 5 minutes. Additional dose was administered at 3 hours after unclamping. Mice were allowed to survive for 24 or 48 hours after surgery. LR, Lactated ringers. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

3 Fig 2 Neurologic scores with ethyl pyruvate (EP) or lactated ringers (LR) treatment (0 = normal function, 6 = complete paraplegia). EP treatment (-▲-) decreased neurologic dysfunction by at 42 hours (∗P < .01) and 48 hours (∗P < .05) compared with LR-treated mice (-●-). The overall incidence of paralysis was decreased by 50%. SE, Standard error of the mean. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

4 Fig 3 Representative images of creysl violet-stained spinal cord cross-sections at 24 and 48 hours following thoracic aortic ischemia reperfusion (TAR). At 24 hours, there was no difference in motor neuron morphology in the lactated ringers (LR)- (A) and the ethyl pyruvate (EP)- (B) treated mice. At 48 hours, LR-treated mice (C) had fewer motor neurons in the anterior horns, while EP treatment (D) showed preservation of motor neuron morphology in the anterior horn of the lumbar segment. E, A representative section from a sham mouse. The graph (F) demonstrates the average motor neuron counted in the anterior horn in the lumbar segment of the spinal cord cross-sections at 100× magnifications after 24 and 48 hours reperfusion. At 48 hours after reperfusion, the LR-treated mice (solid bar) had significantly lower average motor neuron counts compared with the EP-treated group (open bar; P < .01). SE, Standard error of the mean. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

5 Fig 4 Expression of proinflammatory chemokines protein in the spinal cord tissue after thoracic aortic ischemia reperfusion. The expression of the proinflammatory cytokines was measured in the spinal cord tissue. The average keratinocyte chemoattractant (KC) (A; ∗P < .001) and interleukin (IL)-6 (B; ∗∗P < .001) protein level in the lactated ringers (LR)-treated mice was significantly higher at 48 hours after reperfusion compared with the ethyl pyruvate (EP)-treated mice. IL-6 was markedly higher in at 24 hours in the LR-treated group compared with 24 hours after reperfusion in the same group (B, black solid bars; ∗P < .01). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

6 Fig 5 Immunohistochemical detection of Iba-1 in the spinal cord tissue. Representative images of ionized calcium binding adaptor molecule 1 (Iba-1)-stained cross-sections at 24 hours (A and B) or 48 hours (C and D) following thoracic aortic ischemia reperfusion (TAR) or sham surgery (E). A and C, Lactated ringers (LR)-treated mice; B and D, Ethyl pyruvate (EP)-treated mice. There was significantly lower number of Iba-1 stained cells at 24 hours TAR in the EP-treated group (F; P < .05; n = 3). There was no difference in the number of stained cells at 48 hours after TAR (F; P > .05; n = 4). SE, Standard error of the mean. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

7 Fig 6 Bcl-2 and Bax protein levels in ethyl pyruvate (EP)-treated and untreated mice following aortic ischemia vs ischemia reperfusion. Following ischemia alone, EP treatment had higher expression of the antiapoptotic protein Bcl-2 (A; ∗P < .028) compared with lactated ringers (LR) group (n = 4), while Bax protein level was not different at the same time period (B; P = .68). In contrast, by 24 hours after reperfusion, EP-treated mice had a markedly higher Bcl-2 protein level after 24 hours thoracic aortic ischemia reperfusion (TAR) (C; ∗P < .05) and significantly lower Bax protein measured at 48 hours after reperfusion compared with LR-treated mice (D; ∗P < .05). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

8 Fig 7 Immunoflourescent Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining of spinal cord tissue. The upper panel are 50× and 200× magnification of TUNEL green fluorescent positive cells (point white arrows) concentrated in the middle of the spinal cord section and toward the posterior horns of an LR treated mice (A and C). The ethyl pyruvate (EP)-treated mice had few positive cells (B and D). E, This graph summarizes the average TUNEL-positive cells counted per section. There was no difference in the number of TUNEL positive cells at 24 hours after thoracic aortic ischemia reperfusion (TAR; n = 4). However, there was marked increase in the number of positive cells of the LR-treated group compared with EP after 48 hours reperfusion (n = 4; P < .001). Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

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