Microglial Cells And Their Secretion Of Acute High Intraocular Pressure With Retinal Tissue Of Mice

Objective: Glaucoma is a group of optic nerve atrophy and visual field defect as a common feature of the disease, which is the result of a variety of factors. Studies have shown that patients with glaucoma lateral geniculate body, visual cortex and other central visual pathway also exists a corresponding injury, suggesting that glaucoma may be a neurological degenerative disease. The traditional mechanical doctrine and ischemic doctrine can not fully explain the mechanism of glaucoma neural degeneration. In recent years, from the perspective of Immunology, pathogenesis and treatment of glaucoma has become a hot.There is increasing evidence that microglia participated in some of the pathological process of neurodegenerative diseases, microglia are inherent in the immune activity of the retina cells ,in glaucoma retinal ganglion cells play an important role in immune regulation . When the glaucoma damage is increased, the microglia cells and their secretion of matrix metalloproteinases (MMPS) involved in the process of neuronal apoptosis. MMPS is the main protease class of microglia cells secrete, with a variety of extracellular remodeling in normal and pathological processes involved. In which MMP-2, 9 can degrade a variety of extracellular components, ultimately lead to neuronal degeneration and necrosis. In this study, we made high intraocular pressure of rat model and observe microglia cells and their secretion of small matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9) for immunohistochemical staining, after analysis of their expression, it seeks to microglia cells and their secretion of MMP-2, MMP-9 in the genesis and development of glaucoma, the role of lesion and its mechanism explored with a view to the future treatment and prevention of provide experimental evidence of glaucoma. Methods: SD rats of clean grade healthy 30, weighing 200-300g, either sex, experimental animals were randomly divided into two groups, one group (normal group) 6, two groups (high-tension group) 24, 2group was based on perfusion at different times, were randomly divided into 2 hours, 48 hours, 72 hours three groups, each group 8 and eventually abandoned to test the sudden death and substandard eye, each group and the normal control group retain six of the eyes. two Preparation of high intraocular pressure group finished model [1], the 10% chloral hydrate anesthesia rats were killed, complete remove the eye and the intraorbital nerve, micro scissors, micro forceps removal of the conjunctiva, muscle integrity out the optic nerve, PBS ( phosphate buffer) for cleaning, taking less than 0.5cm×0.5cm×0.1cm tissue block, 1 / 3 after 4% paraformaldehyde fixative fixed for HE staining, 2 / 3 by the above-mentioned fixative fixed Across the immune Group Analysis of MMP-2, MMP-9 and microglia cell in the retina of the expression levels, using multi-color cell image analysis management system (Image-Pro Plus 6.0) on the image of the positive part of for average optical density analysis. Experimental data obtained using SPSS13.0 statistical software analysis, comparison between groups using analysis of variance, if significant differences between the two groups with q test.Results: The high intraocular pressure group and the control mice compared with:Morphology:⑴high intraocular pressure group and the normal group, each time the thickness of layers no significant change compared with the normal group.⑵2 hr retinal ganglion cells of the same size, outline the rules, nuclear clear-cell borders clear; 48-hour group of retinal ganglion cells of the same size, slightly irregular contours, nuclear deeply stained, cell borders clear; 72-hour group In addition to the above changes, the outer nuclear layer can see vacuolated changes.Retinal ganglion cell layer of microglia cells and MMP-2, MMP-9's immune chemical analysis:(1) microglia cells in the retina of the expression: The normal group and high intraocular pressure group retinal nerve fiber layer, inner and outer plexiform layer and outer nuclear layer have a small amount of expression, inner nuclear layer of the positive cells in the distribution of the dominant . microglia cells in the normal group the average optical density values was 0.1293±0.0021, high intraocular pressure 2 hours, 48 hours group, 72-hour group, the average optical density values were 0.1672±0.0042,0.3393±0.0077,0.4824±0.0038. Between the two groups: All groups were significantly different, there are statistically significant (P <0.01). high intraocular pressure group and the normal group: With the extension of high intraocular pressure of time, the microglia cells in the retinal ganglion cell layer of expression gradually increased.(2) MMP-2 expression in the retina:MMP-2 expression in normal group is little, high intraocular pressure group of positive MMP-2 particles are stained yellow or brown . the outer plexiform layer and outer nuclear layer made the mainly scattered in the retina,other layers had no significant expression . MMP-2 in the retina of the normal group, the average optical density value was 0.1857±0.0035 high intraocular pressure 2 hours, 48 hours group, 72-hour group, the average optical density values were0.2848±0.0034,0.6464±0.6696,0.8854±0.8069. Between the two groups: All groups were significantly different, there are statistically significant (P <0.01). high intraocular pressure group and the normal group: With the extension of high intraocular pressure of time,MMP-2 in retinal ganglion cell layer of the expression gradually increased.(3) MMP-9 expression in the retina:MMP-9 expression in normal group is little , high intraocular pressure group of positive MMP-9 particles are stained yellow or close to brown yellow, the outer plexiform layer and outer nuclear layer made the mainly scattered in the retina .other layers had no significant expression. MMP-9 in the retina of the normal group, the average optical density value was0.1473±0.037, high intraocular pressure 2 hours, 48 hours group, 72-hour group, the average optical density values were 0.1647±0.0035 ,0.6692±0.0075,0.8771±0.1333. Between the two groups: All groups were significantly different, there are statistically significant (P <0.01). high intraocular pressure group and the normal group: With the extension of high intraocular pressure of time,MMP-9 in retinal ganglion cell layer of the expression gradually increased.Conclusion: 1 High intraocular pressure in rat retinal ganglion cell layer, contour irregularity, nuclear deep-dyed, showing that vacuole-like change.2 High intraocular pressure in rat retinal ganglion cell layer, microglia cells when compared with normal-tension increased.3 High intraocular pressure in rat retinal ganglion cell layer, matrix metalloproteinase -2, when compared with normal-tension were increases.4 High intraocular pressure in rat retinal ganglion cell layer, matrix metalloproteinase -2, when compared with normal-tension were increases5 Microglia cells and MMP-2, MMP-9 involved in the high intraocular pressure in rat retinal ganglion cell layer of the damage repair process.6 Microglia cells and MMP-2, MMP-9 in the late high intraocular pressure in rat retinal ganglion cell layer can enhance the high expression of high intraocular pressure on the optic nerve damage.