Dissertation
Dissertation > Medicine, health > Ophthalmology > Retina and optic nerve diseases > Retinal disease

Protective Action and Mechanism of GBE and Sulforaphen on the Apoptosis of Retinal Nerve Cells in Mice

Author YangYang
Tutor KongLi
School Dalian Medical University
Course Human Anatomy and Embryology
Keywords Ginaton Sulforaphane Retinal light damage Apoptosis
CLC R774.1
Type Master's thesis
Year 2011
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Objective: light can cause retinal cell apoptosis induced retinal outer nuclear layer damage. In this study, mice were light damage to the retina ginaton (GBE) and sulforaphane (SF) heart chamber injection explore GBE and SF to light induced retinal cell apoptosis whether preventive protective effect and its mechanism. Method: 1. Establish a mouse model of light damage: Bal b / c mice were kept dark for 24 hours, the next day into the light box, were irradiated 24h, 48h and 72h were killed, making eye paraffin sections. HE staining of retinal morphology; carried outer nuclear layer and outer nuclear layer thickness measurement number statistics, analysis of different light depending on the length of the cell damage caused; DNA gradient electrophoresis and TUNEL assay, as apoptosis. 2. Bal b / c mice were randomly divided into six groups namely normal control group, the experimental control group (saline), GBE group (GBE-Ⅰ group of 100 ul / secondary; GBE-Ⅱ group 50μl / time, the concentration of 3.5mg/ml), SF group (SF-Ⅰ group was 60μl / secondary; SF-Ⅱ group 30μl / a concentration of 50mg/ml), n = 5, cardiac injection (intravenous replacement), In addition to the normal control group than the five groups of mice were homemade light box light were sacrificed after 72h, conventional preparation eyeball paraffin sections, HE staining morphology and measure the thickness of the outer nuclear layer; RT-PCR detection of cytochrome C and Bak-1 expression; Caspase activity assay kit Caspase-3 activity. Using SPSS13.0 statistical software, single-factor analysis of variance analysis. Results: 1. Mice light damage model: HE staining morphology observed in normal mice retinas clear hierarchy; illumination in each group was significantly thinner outer nuclear layer, retinal photoreceptor cells, the outer segments arranged apparent disorder, unclear boundaries , irregular nuclei. Apoptosis detection and localization: DNA electrophoresis showed that light each group appear DNA laddering, where the light 72h genomic DNA laddering most significant; TUNEL assay showed that apoptosis is located in the outer nuclear layer. 2.GBE light damage to the retina and SF preventive protective effect: morphological observations, experimental control group retinal cells sparsely arranged, the inner and outer sections arranged Ming disorders, swelling, broken, outer nuclear layer was thinner; administration group and The structure of the retina normal control group were better than the experimental control group. Outer nuclear layer thickness measurement results, the normal control group 49.23 ± 2.32μm, experimental control group 40.36 ± 2.17μm; GBE-Ⅰ group, GBE-Ⅱ group of outer nuclear layer thickness was 42.0770 ± 0.8999μm and 40.9411 ± 0.706μm; SF- Ⅰ group, SF-Ⅱ group of outer nuclear layer thickness was 48.2352 ± 2.0447μm and 46.8004 ± 0.6014μm. Statistical analysis showed that the results, GBE-Ⅰ group, GBE-Ⅱ group, SF-Ⅰ group, SF-Ⅱ group and experimental group were significantly difference (P lt; 0.001). 3.GBE light damage to the retina and SF preventive protection mechanisms: GBE-Ⅰ group, GBE-Ⅱ group, SF-Ⅰ group, SF-Ⅱ group and experimental control group, RT-PCR analysis showed cytochrome C and Bak -1 expression were reduced to varying degrees. GBE-Ⅰ group, GBE-Ⅱ group, SF-Ⅰ group, SF-Ⅱ group mRNA levels compared with control group there was significant difference (P lt; 0.01, P lt; 0.05); Caspase-3 activity assay results: GBE - Ⅰ group, GBE-Ⅱ group, SF-Ⅰ group, SF-Ⅱ group and experimental control group, Caspase-3 activity was reduced. Conclusions: 1. Timed lighting can be used to establish mouse retina light damage model; 2.GBE light damage to the retina and SF have a preventive protection. 3.GBE and SF light damage to the retina were associated with protective mechanisms to prevent retinal cell apoptosis and inhibition of protein downregulation.

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