Overexpression of HtrA1 and Exposure to Mainstream Cigarette Smoke Leads to Choroidal Neovascularization and Subretinal Deposits in Aged Mice
Overexpression of HtrA1 and Exposure to Mainstream Cigarette Smoke Leads to Choroidal Neovascularization and Subretinal Deposits in Aged Mice
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DOI：
10.1167/iovs.14-14453
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发表时间：
2014-10-01
影响因子：
4.4
通讯作者：
Iwata, Takeshi
中科院分区：
文献类型：
--
作者：
Nakayama, Mao;Iejima, Daisuke;Iwata, Takeshi
目的.我们使用转基因（Tg）小鼠确定ARMS 2和HtrA 1在脉络膜和视网膜中的功能，并评估主流香烟烟雾对这些小鼠的影响。鸡肌动蛋白启动子（CAG）用于驱动小鼠HtrA 1、人ARMS 2和ARMS 2（A69 S）在小鼠整个身体中表达一年。使用Spectralis HRA+光学相干断层扫描仪（OCT）进行眼底观察。将眼睛切片，用苏木精和伊红（H&E）染色，并用免疫组织化学分析。使用主流吸烟室（INH 06-CIGR 02 A，MIPS）将小鼠暴露于香烟烟雾，30分钟/天，5天/周，持续12周。12周后进行眼底观察和病理学分析。约18.2%的12个月大的HtrA 1 Tg小鼠表现出脉络膜新生血管（CNV）的OCT和阳性免疫染色与抗CD 31和抗纤连蛋白抗体。此外，弹性货车Gieson（EVG）染色显示HtrA 1 Tg小鼠的Bruch膜损伤。在ARMS 2和ARMS 2（A69 S）Tg小鼠中未观察到视网膜变化。暴露于主流香烟烟雾共12周导致野生型（Wt）小鼠的CNV率为7.7%，HtrA 1 Tg小鼠为20%，但对ARMS 2 Tg小鼠没有影响。此外，在暴露于主流香烟烟雾的HtrA 1 Tg小鼠中，在感光细胞和RPE之间观察到异常沉积物。HtrA 1过表达和主流香烟烟雾可独立导致CNV。HtrA 1基因是湿性AMD的一个强危险因素，但并非所有的HtrA 1 Tg小鼠都发生了CNV，这表明CNV的发生取决于多种危险因素。
PURPOSE. We determined the function of ARMS2 and HtrA1 in the choroid and retina using transgenic (Tg) mice and evaluated the effects of mainstream cigarette smoke on these mice.METHODS. The chicken actin promoter (CAG) was used to drive mouse HtrA1, human ARMS2, and ARMS2 (A69S) expression in the entire body of a mouse for one year. Fundus observations were performed with a Spectralis HRA+ optical coherence tomograph (OCT). Eyes were sectioned, stained with hematoxylin and eosin (H&E), and analyzed with immunohistochemistry. Mice were exposed to cigarette smoke for 30 min/d, 5 d/wk for 12 weeks using a mainstream smoking chamber (INH06-CIGR02A, MIPS). After 12 weeks, fundus observations and pathological analyses were performed.RESULTS. Approximately 18.2% of 12-month-old HtrA1 Tg mice exhibited choroidal neovascularization (CNV) by OCT and positive immunostaining with anti-CD31 and anti-fibronectin antibodies. Furthermore, elastic van Gieson (EVG) staining showed Bruch's membrane damage in HtrA1 Tg mice. No retinal changes were observed in ARMS2 and ARMS2 (A69S) Tg mice. A total of 12 weeks of exposure to mainstream cigarette smoke led to CNV rates of 7.7% for wild type (Wt) mice and 20% for HtrA1 Tg mice, but had no effect on ARMS2 Tg mice. In addition, abnormal deposits were observed between photoreceptor cells and the RPE in an HtrA1 Tg mouse exposed to mainstream cigarette smoke.CONCLUSIONS. The HtrA1 overexpression and mainstream cigarette smoke can independently lead to CNV. The HtrA1 gene is a strong risk factor for wet AMD, but not all of the HtrA1 Tg mice developed CNV, suggesting that CNV development depends on multiple risk factors.