Mitochondrial proteomics in experimental autoimmune uveitis oxidative stress.
Mitochondrial proteomics in experimental autoimmune uveitis oxidative stress.
复制标题
DOI：
10.1167/iovs.08-2842
复制
发表时间：
2009-12
影响因子：
4.4
通讯作者：
Rao NA
中科院分区：
文献类型：
--
作者：
Saraswathy S;Rao NA
光感受器线粒体氧化应激是实验性自身免疫性葡萄膜炎的最初病理事件。在这项研究中，作者确定了实验性自身免疫性葡萄膜炎（EAU）早期阶段视网膜线粒体蛋白水平响应氧化应激的变化。制备EAU早期视网膜线粒体组分，并进行二维差异凝胶电泳（2D-DIGE）。切下显示差异表达的蛋白质点，并进行基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）用于肽鉴定。这些蛋白质的水平也通过Western印迹分析证实。这些蛋白质的mRNA表达通过实时PCR证实。TUNEL染色检测细胞凋亡。2D-DIGE分析显示13个蛋白质的差异表达。其中10个蛋白质过表达，包括锰-SOD、αA晶体蛋白、β晶体蛋白，4个蛋白质下调，包括三磷酸腺苷（ATP）合酶、苹果酸脱氢酶和钙视网膜蛋白。Western blot分析证实αA晶体蛋白、βB2晶体蛋白、MnSOD和乌头酸酶水平升高，ATP合酶水平降低。qPCR还证实αA晶体蛋白、βB2晶体蛋白、MnSOD和Hsp 70的表达增加。在这个阶段没有细胞凋亡。EAU早期视网膜中存在视网膜特异性氧化应激相关蛋白沿着ATP合酶的下调，这为应激相关视网膜损伤提供了早期证据。线粒体中高水平的αA和βB2晶体蛋白的存在可能防止早期EAU期间的细胞死亡。
Photoreceptor mitochondrial oxidative stress is the initial pathologic event in experimental autoimmune uveitis. In this study, the authors determined alterations in retinal mitochondrial protein levels in response to oxidative stress during the early phase of experimental autoimmune uveitis (EAU). Retinal mitochondrial fractions during early EAU were prepared and subjected to two-dimensional difference in gel electrophoresis (2D-DIGE). Protein spots showing differential expression were excised and subjected to matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) for peptide identification. Levels of these proteins were also confirmed by Western blot analysis. mRNA expression of these proteins was confirmed by real-time PCR. TUNEL staining was performed to detect apoptosis. 2D-DIGE analysis revealed differential expression of 13 proteins. Ten proteins were overexpressed, including manganese-SOD, αA crystallin, β crystallin, and four proteins were downregulated, including adenosine triphosphate (ATP) synthase, malate dehydrogenase, and calretinin. Increased levels of αA crystallin, βB2 crystallin, MnSOD, and aconitase and decreased levels of ATP synthase were confirmed by Western blot analysis. qPCR also confirmed the increased expression of αA crystallin, βB2 crystallin, MnSOD, and Hsp70. Apoptosis was absent during this phase. The presence of mitochondrial-specific oxidative stress-related proteins in the early EAU retina along with the downregulation of ATP synthase provides early evidence of stress-related retinal damage. The presence of high levels of αA and βB2 crystallin in the mitochondria may prevent cell death during early EAU.
登录
查看更多内容
影响因子：
4.8
作者：
Bornhoevd, Carsten;Vogel, Frank;Reichert, Andreas S.
通讯作者：
Reichert, Andreas S.
影响因子：
3.4
作者：
Duncan, MK;BanerjeeBasu, S;Piatigorsky, J
通讯作者：
Piatigorsky, J
影响因子：
3.4
作者：
Ruiz-Romero, Cristina;Carreira, Vanessa;Blanco, Francisco J.
通讯作者：
Blanco, Francisco J.
影响因子：
3.8
作者：
Lee, Yie Hou;Chung, Maxey C. M.;Boelsterli, Urs A.
通讯作者：
Boelsterli, Urs A.
影响因子：
4.4
作者：
Lifshitz, J;Sullivan, PG;McIntosh, TK
通讯作者：
McIntosh, TK