A 44 mm craniotomy was performed lateral (right side) to the mid-sagittal suture, centered at: AP = 0, ML = 2
A 44 mm craniotomy was performed lateral (right side) to the mid-sagittal suture, centered at: AP = 0, ML = 2.5 from bregma. cause of death and acquired disability in young children [1]. Despite having a high degree of neuroplasticity, young children tend to have poorer outcomes after TBI than adults [2]. Children can also exhibit differential responses to pharmacological interventions including altered bioavailability, metabolism, and drug response [3,4]. It is thus critical that both the effect of TBI and potential therapeutics be investigated in an age-appropriate model [5]. The n-3 long-chain polyunsaturated fatty acids (LC-PUFA) eicosapentaenoic acid (EPA, 20:6n-3) and docosapentaenoic acid (DHA, 22:6n-3), the main constituents in fish oil, are biologically active with many neuroprotective properties. When consumed in the diet or via supplementation, these n-3 fatty acids are incorporated into the phospholipids that form cell membranes where they can alter the physicochemical and membrane-signaling properties of the cell [6,7]. DHA, EPA and their metabolites also have anti-excitotoxic [8], antioxidant [9], anti-apoptotic [10,11], and anti-inflammatory properties [12]. During development, N-3 fatty acids, particularly DHA, accumulate in BRL-54443 the brain during late gestation and early post-neonatal life in humans and rats, a time at which children have a high risk for sustaining traumatic brain injuries [1315]. Additionally, EPA and DHA can be metabolized BRL-54443 into several families of molecules including NPD1, docosanoids, resolvins,etc., which have been shown be neuroprotective through their anti-inflammatory and inflammation-resolving activities [1618]. LC-PUFA can also directly or indirectly modulate gene expression through activation or suppression of cell signaling pathways and transcription factors (e.g., PI3K/Akt, NF-B, PPAR and RXR) [1921]. DHA and EPA, the primary active constituents in fish oil, readily cross the blood-brain barrier [22]. Studies of various neural injury models including TBI and spinal cord injuries in adult animals indicate that fish oil, or DHA or EPA alone produce beneficial BRL-54443 effects [2325]. Furthermore, in a case report, high dose fish oil supplementation (19.2 g/day) was associated with substantial clinical improvement in a young patient with severe, potentially lethal, head trauma [26]. However, the effects of LC-PUFA or fish oil treatment have not been investigated in juvenile brain injury. Accordingly, this study investigated the use of oral fish oil treatment in a juvenile rat model on sensorimotor and biochemical outcomes of TBI, including blood-brain barrier disruption and BRL-54443 levels ofCcl2, Gfap, andMmp9mRNAs, representative key mediators involved in immune cell recruitment, astrocytosis, and blood-brain barrier disruption after TBI. We will show that fish oil resulted in improved functional outcome, at least in part, by limiting disruption of the blood-brain barrier through a mechanism that includes attenuation of TBI-induced expression ofMmp9. == MATERIALS AND METHODS == All experiments were conducted in accordance with the NIH Guide for the Care and Use of Laboratory Animals and were approved by the University of Kansas Medical Center Institutional Animal Care and Use Committee. == Animals, husbandry, and treatment == Long-Evans rats were housed in a temperature- and humidity-controlled facility with a 1410 hour light-dark cycle (on at 06:00 BRL-54443 h) withad libitumaccess to water and chow (Teklad Global diet 2016, Harlan Laboratories, Inc., Indianapolis, IN). The chow contained 4% fat by weight and was formulated with soybean oil resulting in a fatty acid composition of 0.5% 16:0, 0.1% 18:0, 0.7% 18:1n-9, 2% 18:2n-6, 0.1% 18:3n-3. Breeding stock (females 7585 days; male proven breeders; Harlan Laboratories, Inc. Indianapolis, IN) were obtained a minimum of 5 days prior to the beginning of the experiment and Ntrk1 were handled regularly. Litters were culled to eight pups with preference for males on postnatal day (PND) one. Male pups (n = 412/group, depending on endpoint, and each from a different litter) received either a controlled cortical impact (CCI) injury or sham surgery on.