Exploration Of The IBA1, IL-10 And IL-1? CNS Dynamic Expression And Pathogenesis Of EAE Model Of C57BL/6 Mice

Objective:Establish MOG35-55 of C57 BL / 6 mice in experimental autoimmune encephalomyelitis(experimental autoimmune encephalomyelitis, EAE) model, detected early onset(day 7), the peak incidence(day 15) and recovery(Article dynamic changes 28 days) when brain tissue associated with immunomodulatory microglia(IBA1), each period proinflammatory cytokines(such as IL-1?) and anti-inflammatory cytokines(such as IL-10) expression, neurological dysfunction ratings and weight count change. The experiment was discussed and microglia immune-related brain and spinal cord of EAE role within the organization; observe the dynamic changes of EAE brain histopathology, immunology various levels of incidence period; further explore the pathogenesis of EAE, multiple sclerosis, etc. central nervous system demyelinating disease pathogenesis and seek new treatments provide further experimental evidence. Methods:1. 8 to 30 weeks old(body weight 15-20g) C57 BL / 6 female mice were randomly divided into normal control group and EAE group, 21 mice were established EAE model, the day 0 subcutaneously 200 ug mixture of MOG( MOG35-55 and CFA in a vacuum of 1: 1 blend mixture), respectively, at day 0 and day 2 intraperitoneal injection of 200 ng of PTX, normal group injected with 9 metering PBS. In normal mice EAE group immunized and neurological score and take measurements of body weight per day.2. respectively, in the first five days after immunization, 15 days and 28 days of onset phase of their brains were made of continuous paraffin sections and HE staining, Luxol Fast Blue-Eosin staining observed inflammatory cells and myelin depigmentation situation.3.immunohistochemical assay brain spinal cord microglial cell marker(IBA1), the expression of immune factor interleukin 10(IL-10), interleukin-factor(IL-1?) in. Results:1. MOG35-55 model polypeptide induced C57 BL / 6 mice in general and neurological function.MOG-induced mice were immunized average 5.02 ± 1.54 Tian irritability, weight loss, the end of the tension disappears, followed by hind limb weakness, hind limb paralysis and other symptoms. The average clinical score was 2.16 ± 1.42 minutes, 16 days, after which sustained the most severe symptoms of symptoms, no significant remission of symptoms after 28 days decreased slightly, but still mild neurological deficit. Experimental control group and the control group, a significant weight loss and abnormal behavior were not found.2. histopathology(1).HE staining showed: the brain spinal cord parenchyma early group of small blood vessels mainly small veins. See inside the subarachnoid space around a little amount of inflammatory cell infiltration, and often the formation of inflammatory foci small area. Typical small lesions appear inflammatory cells at the peak of the group, showing the brain and spinal cord parenchyma and edge, subarachnoid and small vascular congestion of the choroid plexus of the lateral ventricle, surrounded by a large number of inflammatory cell infiltration of perivascular based vein, the main infiltration of mononuclear cells to form a "cuff" shape change. Mainly lymphocyte infiltration, as small and deeply stained nuclei, the extension of blood vessels. In the hippocampus with varying degrees of nerve cell loss.(2). IFB staining: white matter regions of varying sizes appear blue sheet demyelination, or degeneration. Anterior spinal cord and inner lateral and white matter demyelination were flaky, pale blue stained myelin area, eosin stained red. Myelin sheath loose, disintegration, break, drop out, demyelinating plaque edges uneven structure is not clear. Early discovery group and the peak of the big group of white matter demyelination plaque area, the late group lesions involving cortical gray matter. Conclusions:1.the peak incidence of EAE alleviate the course of which the number of brain tissue inflammation and demyelination degree cells dynamic evolution.2.with the progress of the disease EAE, activation of microglia IBA1, branched forms of these branches by retracting toward ultraamylopectin or amoeboid form transition, microglia showed showed different in the pathogenesis of EAE in the role of disease progression and recovery play an important role.3.disease remission, showing a large number of anti-inflammatory cytokine IL-10 expression, with both pro-inflammatory effects of IL-1? expression is relatively increased, indicating brain and spinal cord tissue IL-1? and IL-10 and microglia synergistic relationship between participation in neuroprotection.