Zhu Shuang, Liu Hua, Sha Haibo, Qi Ling, Gao Dian-Shuai, Zhang Wenbo
Research Center for Neurology, Xuzhou Medical University, Xuzhou, Jiangsu, China; Department of Ophthalmology and Visual Sciences, The University of Texas Medical Branch, Galveston, TX, USA.
Center for Biomedical Engineering, The University of Texas Medical Branch, Galveston, TX, USA.
Exp Eye Res. 2017 Feb;155:1-14. doi: 10.1016/j.exer.2017.01.002. Epub 2017 Jan 5.
Inflammation plays a key role in the pathogenesis of many retinal degenerative diseases related with photoreceptor dysfunction/degeneration. However the involvement of photoreceptor cells in inflammatory reactions is largely unknown as they are not considered as inflammatory cells. In this study, we assessed whether photoreceptor cells can produce CCL2 and CXCL10, two important players in inflammation during endoplasmic reticulum (ER) stress. After photoreceptor 661 W cells were treated with ER stress inducer thapsigargin (TG), induction of ER stress increased CXCL10 and CCL2 expression at both mRNA and protein levels, which was significantly blocked by an ER stress blocker 4-phenylbutyrate. ER stress contains three pathways: PERK, ATF6 and IRE1α. Knockdown of PERK attenuated TG-induced CXCL10 and CCL2 mRNA expression, associated with significant decreases in phosphorylation of NF-κB RelA and STAT3. In contrast to PERK, knockdown of XBP1, which is activated by IRE1α-mediated splicing, robustly enhanced TG-induced CXCL10 and CCL2 expression and phosphorylation of NF-κB RelA and STAT3. Blockade of NF-κB or STAT3 markedly diminished TG-induced CXCL10 and CCL2 expression. The specific roles of PERK and XBP1 in CXCL10 and CCL2 expression were further investigated by treating photoreceptor cells with advanced glycation end products (AGE) and high glucose (HG), two of the major contributors to diabetic complications. Similarly, AGE and HG induced CXCL10 and CCL2 expression in which PERK was a positive regulator while XBP1 was a negative regulator. These studies suggest that photoreceptors may be involved in retinal inflammation by expressing chemokines CXCL10 and CCL2. PERK and IRE1α/XBP1 in the unfolded protein response differentially regulate the expression of CXCL10 and CCL2 likely through modulation of ER stress-induced NF-κB RelA and STAT3 activation.
炎症在许多与光感受器功能障碍/退化相关的视网膜退行性疾病的发病机制中起关键作用。然而,光感受器细胞在炎症反应中的参与情况在很大程度上尚不清楚,因为它们不被视为炎症细胞。在本研究中,我们评估了光感受器细胞是否能产生CCL2和CXCL10,这两种在内质网(ER)应激期间炎症中的重要参与者。在用ER应激诱导剂毒胡萝卜素(TG)处理光感受器661W细胞后,ER应激的诱导在mRNA和蛋白质水平上均增加了CXCL10和CCL2的表达,这被ER应激阻断剂4-苯基丁酸显著阻断。ER应激包含三条途径:PERK、ATF6和IRE1α。敲低PERK可减弱TG诱导的CXCL10和CCL2 mRNA表达,同时NF-κB RelA和STAT3的磷酸化显著降低。与PERK相反,敲低由IRE1α介导的剪接激活的XBP1,可强烈增强TG诱导的CXCL10和CCL2表达以及NF-κB RelA和STAT3的磷酸化。阻断NF-κB或STAT3可显著减少TG诱导的CXCL10和CCL2表达。通过用晚期糖基化终产物(AGE)和高糖(HG)处理光感受器细胞,进一步研究了PERK和XBP1在CXCL10和CCL2表达中的具体作用,AGE和HG是糖尿病并发症的两个主要促成因素。同样,AGE和HG诱导了CXCL10和CCL2表达,其中PERK是正调节因子,而XBP1是负调节因子。这些研究表明,光感受器可能通过表达趋化因子CXCL10和CCL2参与视网膜炎症。未折叠蛋白反应中的PERK和IRE1α/XBP1可能通过调节ER应激诱导的NF-κB RelA和STAT3激活来差异调节CXCL10和CCL2的表达。