El Mathari Brahim, Sene Abdoulaye, Charles-Messance Hugo, Vacca Ophélie, Guillonneau Xavier, Grepin Claudine, Sennlaub Florian, Sahel José-Alain, Rendon Alvaro, Tadayoni Ramin
Institut de la Vision/INSERM/UPMC, Univ Paris 06/CNRS/CHNO des Quinze-Vingts, Paris, France, Sanofi Fovea-Ophthalmology, Paris, France.
Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, Saint Louis, MO, USA.
Hum Mol Genet. 2015 Jul 15;24(14):3939-47. doi: 10.1093/hmg/ddv132. Epub 2015 Apr 21.
We have previously shown that the deletion of the dystrophin Dp71 gene induces a highly permeable blood-retinal barrier (BRB). Given that BRB breakdown is involved in retinal inflammation and the pathophysiology of many blinding eye diseases, here we investigated whether the absence of Dp71 brings out retinal vascular inflammation and vessel loss by using specific Dp71-null mice. The expression of vascular endothelial growth factor (VEGF), quantified by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay methods, was higher in the retina of Dp71-null mice than in wild-type mice. In contrast, no differences were observed in VEGFR-2 and tumor necrosis factor-α expression. Moreover, mRNA expression of water channel, aquaporin 4 (AQP4) was increased after Dp71 deletion. The Dp71 deletion was also associated with the overexpression of intercellular adhesion molecule 1, which is expressed on endothelial cells surface to recruit leukocytes. Consistent with these findings, the total number of adherent leukocytes per retina, assessed after perfusion with fluorescein isothiocyanate-conjugated concanavalin A, was increased in the absence of Dp71. Finally, a significant increase in capillary degeneration quantified after retinal trypsin digestion was observed in mice lacking Dp71. These data illustrate for the first time that the deletion of Dp71 was associated with retinal vascular inflammation, vascular lesions with increased leukocyte adhesion and capillary degeneration. Thus, dystrophin Dp71 could play a critical role in retinal vascular inflammation disease, and therefore represent a potential therapeutic target.
我们之前已经表明，缺失肌营养不良蛋白Dp71基因会诱导血视网膜屏障（BRB）高度通透。鉴于BRB破坏与视网膜炎症以及许多致盲眼病的病理生理学有关，在此我们使用特定的Dp71基因敲除小鼠，研究了Dp71缺失是否会引发视网膜血管炎症和血管丧失。通过定量聚合酶链反应和酶联免疫吸附测定方法定量的血管内皮生长因子（VEGF）的表达，在Dp71基因敲除小鼠的视网膜中高于野生型小鼠。相比之下，在VEGFR-2和肿瘤坏死因子-α表达方面未观察到差异。此外，水通道蛋白4（AQP4）的水通道mRNA表达在Dp71缺失后增加。Dp71缺失还与细胞间黏附分子1的过表达有关，该分子在内皮细胞表面表达以募集白细胞。与这些发现一致，在用异硫氰酸荧光素偶联的伴刀豆球蛋白A灌注后评估的每个视网膜中黏附白细胞的总数，在缺乏Dp71时增加。最后，在缺乏Dp71的小鼠中，视网膜胰蛋白酶消化后定量的毛细血管变性显著增加。这些数据首次表明，Dp71的缺失与视网膜血管炎症、白细胞黏附增加的血管病变和毛细血管变性有关。因此，肌营养不良蛋白Dp71可能在视网膜血管炎症疾病中起关键作用，因此代表一个潜在的治疗靶点。
