The Express Of NF-κB And The Molecular Mechanism Of Cholecystokinin-8 On Anti-Inflammation Of Acute Spinal Cord Injury In Rars

Inflammatory responses are a major component of secondary injury and play a central role in mediating the pathogenesis of acute and chronic spinal cord injury (SCI). The nuclear factor-kappa B (NF- K B) is a primary transcription factor for the transcription activation of a variety of cytokinin and genes regulating inflammatory. The activation of NF- K B made inflammation genes over expression including tumor necrosis factor-alpha (TNF-a) and Interleukin-lbeta(IL-l). But the expression of NF-KB and inhibitory subunit of NF- K. B(I K B) in SCI is not very clear, exespecially the expression of I K B in spinal cells .Glial fibrillary acidic protein (GFAP) is an important role of SCI .The increase of the number and expression of GFAP-positive cells was correlated to the inflammatory responses and functional recovery of the spinal cord. We used a contusion model in rats to initiate the early biochemical and molecular changes with acute inflammation after SCI and examined the expression of activated NF- K BN I K B and GFAP with immunohistochemistry western blot analyses and electrophoretic mobility shift assays (EMS A). In order to search the new medicine of relieving inflammatory responses we injected different dose of CCK-8 (Cholecystokinin-octapeptide) to rats before injury and examined the expression of NF- K B^ I K B, GFAP.1 Expression of activated NF- K B after acute traumatic SCI.The purpose is observe the expression of NF- K B at cytochylema and nucleolus of SD rat in ASCI. All the animal were divided into control group(l 8) and injury group(72). 1% nembutal was injected in anaesthesia. We applied OHBA SIKI spinal cord beat equipment and finished contusionmodel with animal of injury group. Only laminectomy of Tg 9 were finised to control group. All the animal were killed at different time point which included UK 6rK 12rK 24h. 72h and 5d. Immunohistochemistryx western blot analysis^ EMS A experiments were finished . The expression of NF-K B at every time point was recorded .At Ih after ASCI, There was angiorrhexis at area of contusion of spinal cord. Lots of agminated akaryocyte and plasma could be found at dorsal horn. At 6h after ASCI, nerve cells and astrocyte showed obvious edema. At 12h after SCI, a well defined hemorrhagic zone was observed within the central gray matter of the spinal cord. Within the epicenter of the contusion, gray matter structures appeared necrotic, with polymorphonuclear leukocytes (PMNL) invading the injured parenchyma. Necrotic neurons contained pyknotic nuclei surrounded by an eosinophilic cytoplasm. Severely damaged white matter tracts appeared swollen and edematous. In gray areas bordering the contusion, selective neuronal necrosis was observed within parenchyma containing swollen astrocytic cell bodies. In addition, petechial hemorrhages were observed throughout the gray matter and white matter tracts remote from the contusion. At 24 and after contusion injury, Photomicrographs show necrosis, infiltration of leukocytes, and white matter vacuolization. Severe vacuolization of the white matter was observed after SCI. At 72 hr after SCI, large numbers of macrophages were observed in the white matter. Macrophages and PMNLs were present at the gray/white interface. By 3 d after spinal cord trauma, the lesion remained well defined, and evidence of hemorrhage was observed. These foamy macrophages were seen within necrotic gray and white matter tracts. PMNLs also were dispersed throughout the necrotic areas . By 5 d after spinal cord trauma within areas adjacent to the injury, petechial hemorrhages were detected, and blood vessels demonstrated red blood cell stasis. Blood vessels also contained luminal leukocytes.In order to discuss the express and rule of NF- K B in the spinal cord, we carried out Immunohistochemistry research. In control group(90.1+3.1), Little immunoreactivity of NF- K B can be found. DAB staining was absent when the primary antibody was omitted or replaced with a control antibody of an identical isotype. At 1 h postinjury, NF- K B immunorea...