Department of Ophthalmology, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan.
Lab Invest. 2011 Sep;91(9):1277-90. doi: 10.1038/labinvest.2011.101. Epub 2011 Jun 27.
Intraocular neovascularization is the leading cause of severe visual loss and anti-vascular endothelial growth factor (VEGF) therapy is currently performed for choroidal neovascularization (CNV). Despite its potent anti-angiogenic effect, there are concerns about its long-term safety. Non-steroidal anti-inflammatory drugs (NSAIDs) are common therapeutic agents used for treating inflammatory diseases, and their anti-stress effects are attracting attention now. We studied the effects of topical NSAIDs on CNV, focusing on anti-stress proteins. Cultured retinal pigment epithelium (RPE) cells were treated with NSAIDs: bromfenac, indomethacin, or vehicle control. Transcription factor NF-E2-related factor 2 (Nrf2) and its downstream anti-oxidant protein heme oxygenase (HO)-1 were assessed using western blot and immunohistochemistry. As a result, NSAIDs induced translocation of Nrf2 into the nucleus and the robust expression of HO-1 in a dose- and time-dependent manner. Flow cytometric analysis revealed that bromfenac inhibited H(2)O(2)-induced apoptosis in cultured RPE cells. Next, we studied the effects of topical bromfenac on laser-induced CNV model in rat. The expressions of Nrf2 and HO-1, infiltrations of ED-1-positive macrophages at CNV lesions and size were analyzed. VEGF in the ocular fluid of these rats was also measured using enzyme-linked immunosorbent assay. Rats administered an inhibitor of HO-1 stannic mesoporphyrin (SnMP) were also studied. The results showed that topical bromfenac led to translocation of Nrf2 and induction of HO-1 in CNV lesions and that the number of infiltrating macrophages at the CNV lesion decreased. The sizes of CNV lesions were significantly smaller in bromfenac-treated rats than control CNV, and the effects were diminished by SnMP. VEGF increased in the ocular fluid after laser treatment and was inhibited by bromfenac and SnMP canceling these effects. NSAIDs inhibit CNV through the novel anti-stress protein HO-1-dependent pathway, indicating its potential therapeutic value for various intraocular angiogenic diseases including CNV.
眼内新生血管是导致严重视力丧失的主要原因，目前抗血管内皮生长因子 (VEGF) 治疗用于脉络膜新生血管 (CNV)。尽管其具有强大的抗血管生成作用，但人们对其长期安全性仍存在担忧。非甾体抗炎药 (NSAIDs) 是用于治疗炎症性疾病的常用治疗药物，其抗应激作用现在受到关注。我们研究了局部 NSAIDs 对 CNV 的影响，重点关注抗应激蛋白。用 NSAIDs（溴芬酸、吲哚美辛或载体对照）处理培养的视网膜色素上皮 (RPE) 细胞。使用 Western blot 和免疫组织化学评估转录因子 NF-E2 相关因子 2 (Nrf2) 和其下游抗氧化蛋白血红素加氧酶 (HO)-1。结果，NSAIDs 诱导 Nrf2 向核内转移，并以剂量和时间依赖的方式强烈表达 HO-1。流式细胞术分析显示溴芬酸抑制了培养的 RPE 细胞中 H2O2 诱导的凋亡。接下来，我们研究了局部溴芬酸对大鼠激光诱导的 CNV 模型的影响。分析了 CNV 病变中 Nrf2 和 HO-1 的表达、ED-1 阳性巨噬细胞的浸润程度和大小。还使用酶联免疫吸附试验测量了这些大鼠眼内液中的 VEGF。还研究了给予 HO-1 抑制剂锡原卟啉 (SnMP) 的大鼠。结果表明，局部溴芬酸导致 Nrf2 易位并诱导 CNV 病变中 HO-1 的表达，CNV 病变中浸润的巨噬细胞数量减少。与对照 CNV 相比，溴芬酸治疗的大鼠 CNV 病变明显较小，而 SnMP 则减弱了这种作用。激光治疗后眼内液中的 VEGF 增加，被溴芬酸和 SnMP 抑制，取消了这些作用。NSAIDs 通过新型抗应激蛋白 HO-1 依赖性途径抑制 CNV，表明其对包括 CNV 在内的各种眼内血管生成性疾病具有潜在的治疗价值。
