Dept. of Ophthalmology and Visual Sciences, Univ. of Nebraska Medical Center, Omaha, 68198-5840, USA.
Am J Physiol Cell Physiol. 2011 Oct;301(4):C954-67. doi: 10.1152/ajpcell.00061.2011. Epub 2011 Jun 15.
The multifunctional cytoprotective protein peroxiredoxin 6 (Prdx6) maintains cellular homeostasis and membrane integrity by regulating expression of intracellular reactive oxygen species (ROS) and phospholipid turnover. Using cells derived from targeted inactivation of Prdx6 gene or its depletion by RNA interference or aging, we showed that Prdx6 deficiency in cells evoked unfolded protein response (UPR), evidenced by increased expression or activation of proapoptotic factors, CHOP, ATF4, PERK, IRE-α and eIF2-α and by increased caspases 3 and 12 processing. Those cells displayed enhanced and sustained expression of endoplasmic reticulum (ER) stress-related chaperon proteins, Bip/glucose-regulated protein 78, calnexin, and calreticulin. Under cellular stress induced by hypoxia (1% O(2) or CoCl(2) treatment) or tunicamycin, Prdx6-deficient cells exhibited aberrant activation of ER stress-responsive genes/protein with higher expression of ROS, and died with apoptosis. Wild-type cells exposed to tunicamycin or hypoxia remained relatively insensitive with lower expression of ROS and ER-responsive genes than did Prdx6-deficient cells, but upregulation of ER stress responsive proteins or chaperones mimicked the UPR response of Prdx6-deficient or aging cells. Expression of Prdx6 blocked ER stress-induced deleterious signaling by optimizing physiologically aberrant expression of ER stress responsive genes/proteins in Prdx6-deficient cells or cells facing stressors, and rescued the cells from apoptosis. These findings demonstrate that impaired homeostasis and progression of pathogenesis in Prdx6-deficient lens epithelial cells or in aging cells should be blocked by a supply of Prdx6. The results provide a new molecular basis for understanding the etiology of several age-associated degenerative disorders, and potentially for developing antioxidant Prdx6-based therapeutics.
多功能细胞保护蛋白过氧化物酶 6(Prdx6)通过调节细胞内活性氧(ROS)和磷脂周转率来维持细胞内稳态和膜完整性。使用靶向敲除 Prdx6 基因或通过 RNA 干扰使其耗尽或衰老的细胞,我们表明 Prdx6 缺乏会引发未折叠蛋白反应(UPR),证据是促凋亡因子 CHOP、ATF4、PERK、IRE-α和 eIF2-α的表达或激活增加,以及 caspase 3 和 12 的加工增加。这些细胞显示出增强和持续表达内质网(ER)应激相关伴侣蛋白,Bip/葡萄糖调节蛋白 78、钙连蛋白和钙网蛋白。在缺氧(1% O(2)或 CoCl(2)处理)或衣霉素诱导的细胞应激下,Prdx6 缺陷细胞表现出 ER 应激反应基因/蛋白的异常激活,伴随着 ROS 表达增加,并通过凋亡死亡。暴露于衣霉素或缺氧的野生型细胞相对不敏感,ROS 和 ER 反应基因的表达低于 Prdx6 缺陷细胞,但 ER 应激反应蛋白或伴侣的上调模拟了 Prdx6 缺陷或衰老细胞的 UPR 反应。Prdx6 的表达通过优化 Prdx6 缺陷细胞或面临应激的细胞中 ER 应激反应基因/蛋白的生理异常表达,阻断 ER 应激诱导的有害信号,从而拯救细胞免于凋亡。这些发现表明,Prdx6 缺陷的晶状体上皮细胞或衰老细胞中稳态受损和发病机制进展应通过供应 Prdx6 来阻断。该结果为理解几种与年龄相关的退行性疾病的病因提供了新的分子基础,并为开发基于抗氧化剂 Prdx6 的治疗方法提供了潜力。