Analyse the mechanism of choroidal neovascularization (CNV) induced by oxidized LDL receptor and develope the therapy against CNV

1. 在黄斑下手术中，巨噬细胞积聚在脉络膜新生血管膜上。其中一部分被A类清道夫受体（SR-A）抗体免疫染色呈阳性，SR-A对动脉硬化很重要。除老年性黄斑变性外，新生血管膜也含有SR-A阳性巨噬细胞。可能清道夫受体在脉络膜新生血管形成中起重要作用。以野生型和LOX-1基因敲除（LOX-1 KO）小鼠为实验对象，制备激光诱导脉络膜新生血管（CNV）实验模型。暴露后诱导LOX-1 mRNA表达。野生小鼠的前体基质金属蛋白酶（MMPs）蛋白水平升高，MMPs被激活，而在LOX-1 KO小鼠中，前体MMPs的表达和激活被抑制。野生型小鼠在激光照射区域内的新生血管检出率为93%，而LOX-1 KO小鼠的新生血管检出率为43%。这些数据提示LOX-1可能促进脉络膜新生血管的形成。3-羟基-3-甲基戊二酰辅酶A（HMG-CoA）还原酶抑制剂（pitavastatin）具有抑制动脉粥样硬化中高脂血症和斑块形成的作用，为了评估其对CNV的影响，我们在大鼠身上建立了激光诱导CNV的实验模型。匹伐他汀处理大鼠激光照射区荧光渗漏明显减少。经匹伐他汀处理的水稻CNV面积和厚度均较对照显著减小。说明吡伐他汀治疗剂量能有效抑制实验性大鼠CNV。为了筛选人眼内液中所含的蛋白质，应用蛋白质组学分析获得了二维电泳图谱；从白内障手术中切除的体液中提取。采用矩阵-a辅助激光解吸/电离飞行时间质谱法和数据库检索技术对银染蛋白斑点进行了提取和鉴定。白蛋白、异白蛋白、转铁蛋白、载脂蛋白、维生素D结合蛋白、转威胁蛋白和抗胰蛋白酶是人水溶体液中的主要蛋白

1. Macrophage cells accumulated in choroidal neovascular membranes excised surgically during submacular operation. A portion of them was positively immunostained with antibodies against Class-A scavenger receptors (SR-A) that were important for aterosclerosis. Neovascular membranes other than Age-related macular degeneration also contained SR-A positive macrophage cells. It is possibile that scavenger receptors have an important role for the choroidal neovascularization.2. The experimental models of laser-induced choroidal neovascularization (CNV) were prepared in wild type and LOX-1 gene knockout (LOX-1 KO) mice. The expression of LOX-1 mRNA was induced after exposure. The protein level of precursor matrix metalloproteinases (MMPs) was increased and MMPs were activated in wild mice, but in LOX-1 KO mice, the expression of precursor MMPs and activation of MMPs were suppressed. In wild type mice neovascularity within the laser-exposed area was detected with 93 %, but in LOX-1 KO mice it … More was reduced with 43 %. These data suggest that LOX-1 may promote the choroidal neovascularization.3. To evaluate the effect of the administration of the 3-hydroxy-3-methylglutaryl co-enzyme A(HMG-CoA) reductase inhibitors (pitavastatin) on CNV, that was known to suppress hyperlipidemia and plaque formation in atherosclerosis, the experimental models of laser-induced CNV was created in rats. Pitavastatin-treated rats had significantly less fluorescence leakage in laser-exposed area. Both the area and the thickness of CNV in pitavastatin-treated rais were significantly reduced compared with control. These indicate that the therapeutic dose of pitavastatin effectively suppressed experimental CNV in rats.4. In order to screen the protein cotained in the human intraocular fluid, the proteomics analysis was applied to get the two-dimensional electrophoresis profile; from the aqueous humors excised surgically during cataract operations. The silver-staining protein spots were picked up and identified by matrix-a: sisted laser desorption/ionization time-of-flight mass spectrometry and data base searching. Alubmin, alloalubmin, transferrin, apolipoprotein, vitamin D binding protein, transthretin, and antitrypsin were major proteins in the human aqueous humors Less