Lin Wen, Xu Guoxing
Department of Ophthalmology, The First Affiliated Hospital of Fujian Medical University, 20 Chazhong Road, Fuzhou City, 350005, China.
Fujian Institute of Ophthalmology, Fuzhou City, China.
In Vitro Cell Dev Biol Anim. 2018 May;54(5):355-365. doi: 10.1007/s11626-018-0243-9. Epub 2018 Mar 21.
Increasing evidence has demonstrated that excessive blue-light (BL) with high photochemical energy and phototoxicity could induce apoptosis in retinal pigment epithelium (RPE) cells. RPE apoptosis leads to retina damage and further aggravate age-related macular degeneration (ARMD). Because of their neuroprotective, plasticity, and immunomodulatory ability, bone marrow mesenchymal stem cells (BMSCs) are recognized for retinal neuroprotection. RPE cells possess ciliary neurotrophic factor (CNTF) receptor complexes and can respond to CNTF; hence, we investigated the effects of BMSCs over-expressing CNTF on BL-injured RPE cells. BL-injured RPE cells were co-cultured with CNTF-BMSCs and GFP-BMSCs for 24 and 48 h. Superoxide dismutase and malondialdehyde assays were conducted to examine the effects of CNTF-BMSCs on the oxidative stress of RPE cells. VEGF protein secretion by RPE was determined by ELISA, and western blotting analysis was used to determine apoptotic protein expression and autophagic flux. Immunofluorescence was used to demonstrate the relationship between autophagy and apoptosis. We found that CNTF-BMSCs enhanced antioxidant capacity, decreased VEGF secretion, promoted autophagic flux, and inhibited apoptosis in BL-injured RPE cells, compared to GFP-BMSCs. Our findings suggest that CNTF over-expression enhances the protective effects of BMSCs on RPE cells, thus indicating subretinal-transplantation of CNTF-BMSCs may be a promising therapy for BL-injured retina.
越来越多的证据表明，具有高光化学能量和光毒性的过量蓝光（BL）可诱导视网膜色素上皮（RPE）细胞凋亡。RPE细胞凋亡会导致视网膜损伤，并进一步加重年龄相关性黄斑变性（ARMD）。骨髓间充质干细胞（BMSCs）因其神经保护、可塑性和免疫调节能力而被认为对视网膜具有神经保护作用。RPE细胞具有睫状神经营养因子（CNTF）受体复合物，能够对CNTF作出反应；因此，我们研究了过表达CNTF的BMSCs对蓝光损伤的RPE细胞的影响。将蓝光损伤的RPE细胞与CNTF-BMSCs和GFP-BMSCs共培养24小时和48小时。进行超氧化物歧化酶和丙二醛检测，以检查CNTF-BMSCs对RPE细胞氧化应激的影响。通过ELISA测定RPE分泌的VEGF蛋白，并使用蛋白质免疫印迹分析来确定凋亡蛋白表达和自噬通量。采用免疫荧光法来证明自噬与凋亡之间的关系。我们发现，与GFP-BMSCs相比，CNTF-BMSCs增强了抗氧化能力，减少了VEGF分泌，促进了自噬通量，并抑制了蓝光损伤的RPE细胞凋亡。我们的研究结果表明，CNTF的过表达增强了BMSCs对RPE细胞的保护作用，因此表明视网膜下移植CNTF-BMSCs可能是治疗蓝光损伤视网膜的一种有前景的疗法。
