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Establishment of Animal Model for Experimental Autoimmune Encephalomyelitis and Matrix Metalloproteinase-9 Expression on EAE Rats

Author: DuLi
Tutor: CaoXia
School: Taishan Medical College
Course: Internal Medicine
Keywords: Experimental allergic encephalomyelitis Multiple sclerosis Wistar rats Blood-brain barrier MMP-9 Inflammatory factors
CLC: R-332
Type: Master's thesis
Year: 2010
Downloads: 19
Quote: 0
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Abstract


Objective 1, the establishment of experimental allergic encephalomyelitis (EAE) model. The pathological changes observed the EAE rat behavior and central nervous system (CNS), detection CNS matrix metalloproteinase-9 (MMP-9) distribution and expression. 3, for EAE, the treatment of multiple sclerosis (MS), to provide a new idea. Female Wistar rats were randomly divided into the model group (EAE group) and control group (CFA group). Given of EAE group of rats limbs footpad intradermal injection induced emulsion guinea pig spinal cord homogenate - complete Freund's adjuvant (GPSCH-CFA) 0.4 ml of the dorsum of the foot of the left hind rat EAE subcutaneous injection of pertussis vaccine (BPV) 0.2ml, preparation model. Given the CFA group was injected with the same amount of saline and complete Freund's adjuvant (NS CFA). All rats were neurological score, measuring changes in body weight, the dynamic observation EAE rat brain and spinal cord tissue pathological changes and the expression of MMP-9. 1 behavior change: of EAE rats in GPSCH-CFA injection after 8-12 days in succession incidence, mainly as the double hind limb weakness, paralysis, the tail muscle tension reduce disappeared rats forelimbs forward crawling, sword part of the body below the ground in the sudden double hindlimb paralysis to drag in face-paw, some rats accompanied incontinence. Peaked 3-5 days after the onset of clinical manifestations, severe the forelimb muscle tension lowering, quadriplegia, moribund state, about seven days after the onset of EAE symptoms began to recover, the disease faster recovery. The EAE rats body weight compared with the CFA group decreased neurological score was significantly higher compared with the CFA group, the highest score of 5. CFA rats were not EAE symptoms normal circumstances with normal animals, activities, normal diet, weight gradually increased. 2, the pathological changes: light microscope, as shown in the brain and spinal cord parenchyma inflammatory cell infiltration, mainly lymphocytes, mainly distributed in small blood vessels around the cuff-like change was typical to white matter and gray at the junction of the more obvious. KB staining showed that the white matter of the small veins around the peak incidence of brain and spinal cord parenchyma large areas of demyelination and the convalescent visible remyelination phenomenon. Bodian staining showed axonal reserved throughout the course of the disease is relatively good. Immunohistochemical staining showed that the the EAE rats incidence of early brain, spinal cord within a large number of MMP-9 positive cells infiltration, typical MMP-9 positive cells cytoplasm tan, mainly lymphocytes, monocytes, 14 days after the onset of positive The cells begin to reduce expression, no significant expression of MMP-9 in January. Electron microscope of EAE rat brain, spinal cord substance of capillary endothelial cells, mitochondrial swelling, more common stenosis atresia, myeloid bodies, nerve cells and glial cells within the endoplasmic reticulum and mitochondrial swelling myelinated nerve fibers axonal microfilaments microtubules sparse sheath the distort or level separation. Control group, no significant histopathological changes. Conclusion 1 guinea pig spinal cord homogenate - complete Freund's adjuvant (GPSCH-CFA) supplemented pertussis vaccine (BPV) induced in Wistar rats, can be successfully prepared EAE model. 2 of EAE rats pathological changes mainly the cerebrospinal parenchyma the small perivascular inflammatory cell infiltration, was typical of the \relatively intact, and the change is consistent with the pathology of human multiple sclerosis (MS). 3, MMP-9 expression in the EAE group rat cerebrospinal parenchyma increased significantly, and is consistent with the severity of the disease, showed that MMP-9 play a role in EAE pathogenesis.

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