F TMP on spinal cord I/R injury are related to scavenging free radicals and suppressing apoptotic pathways. In this study, the authors investigated the effect of TMP on the neurologic function, biochemical and histopathological changes and purchase Stattic studied its impact on expression of proand anti-apoptotic proteins as well as the numbers of apoptotic cells following spinal cord I/R injury in rabbits.the onset of reperfusion. Control animals underwent standard aortic occlusion and intravenous injection of 0.9 sodium chloride under conditions identical to the TMP injection. Sham operated animals subjected to operative dissections without aortic occlusion. Each group of animals was divided into four experimental subgroups: group A for Biochemical analysis (n = 3), group B for hematoxylin and eosin staining (H E), Terminal Deoxynucleotidyltransferase-Mediated dUTP Nick End-Labeling (TUNEL) staining and immunohistochemistry (n = 3), group C for electron microscopy (n = 2), group D for Western blot assay (n = 4). The rabbit model of spinal cord I/R injury was established according to Savas’discription [14]. Briefly, after sterile preparation, a 10-cm midline incision was performed. Following anticoagulation with 400 unit’s heparin, the abdominal aorta was cross-clamped at the level just inferior to the origin of the left renal artery and at the level of aortic bifurcation for 30 min. Reperfusion was initiated by removal of the occlusion and lasted 48 h. The abdomen was then closed.Neurologic evaluation Neurological function was observed at the 24th and 48th hour after reperfusion according to Johnson’s score[15].0: Hind-limb paralysis;MethodsAll experimental protocols were approved by our Institutional Committee on Animal Research, and were carried out in accordance with the National Institutes of Health guidelines for animal use and care (National Institutes of Health publication no. 96- 23, revised 1996). Experiments were performed on 36 adult male New Zealand White rabbits (provided by Experimental Animal Center of the Xi’an Jiaotong University) weighing 2.5 to 3.0 kg. The animals were initially anaesthetised with pentobarbital sodium (30 mg/kg IV, sigma, USA, NO: 20030709), followed by a half-dose as required during surgical procedure. No animals received hemodynamic or ventilatory support. PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26866270 The left ear vein was cannulated with a 24-gauge catheter for intravenous drug administration. The right femoral artery was catheterized for blood pressure and heart rate monitoring (Spacelab, USA, model 90206A). Arterial blood was sampled for determination of blood gases (AVL-2, Switzerland) and blood glucose (One Touch II, USA). The rectal body temperature was maintained close to 38 with the aid of a heating pad during the study.Experimental groups and Animal models Rabbits were randomly assigned to 3 groups (n = 12 each). In the TMP group, TMP (30 mg/kg) (Changzhou Pharmacological Co., China, NO: 99091401) was injected via ear vein 30 min before aortic clamping and at1: Severe paraparesis; 2: Functionalmovement, no hop; 3: Ataxia, disconjugate hop; 4: Minimal ataxia; 5: Normal function. Two individuals without knowledge of the treatment graded neurological function independently.Histological study The animals were euthanized by intravenous administration of a high concentration of pentobarbital at the 48th hour and the spinal cords were quickly removed. The spinal cords were immersed in 4 paraformaldehyde in 0.1 mol/l phosphate buffer and stored at 4 for.
