Tenascin-C promotes angiogenesis in fibrovascular membranes in eyes with proliferative diabetic retinopathy
Tenascin-C promotes angiogenesis in fibrovascular membranes in eyes with proliferative diabetic retinopathy
复制标题
DOI：
--
复制
发表时间：
2016-04
期刊：
影响因子：
2.2
通讯作者：
Yoshiyuki Kobayashi;S. Yoshida;Yedi Zhou;Takahito Nakama;K. Ishikawa;Mitsuru Arima;S. Nakao;Y. Sassa;A. Takeda;T. Hisatomi;Y. Ikeda;A. Matsuda;K. Sonoda;T. Ishibashi
中科院分区：
文献类型：
--
作者：
Yoshiyuki Kobayashi;S. Yoshida;Yedi Zhou;Takahito Nakama;K. Ishikawa;Mitsuru Arima;S. Nakao;Y. Sassa;A. Takeda;T. Hisatomi;Y. Ikeda;A. Matsuda;K. Sonoda;T. Ishibashi
我们之前已经证明tenascin-C在增殖性糖尿病视网膜病变（PDR）患者的纤维血管膜（fvm）中高度表达。然而，其在fvm发病机制中的作用尚未确定。本研究旨在探讨tenascin-C在fvm形成和血管生成中的作用。方法采用夹心酶联免疫吸附法测定PDR患者玻璃体标本中tenascin-C的水平，并以黄斑孔为对照。采用免疫组化方法检测PDR患者fvm中腱素c、α-平滑肌肌动蛋白（SMA）、CD34、胶质纤维酸性蛋白（GFAP）和整合素α v的位置。我们还测量了tenascin-C mRNA和蛋白在体外受白细胞介素(IL)-13刺激的血管平滑肌细胞（VSMCs）中的表达。在培养的人视网膜内皮细胞（HRECs）中测定tenascin-C对细胞增殖、迁移和管形成的影响。结果PDR患者玻璃体中tenascin-C的平均水平明显高于黄斑裂孔患者（p<0.001）。PDR患者fvm的双免疫荧光分析显示，tenascin-C与α-SMA、CD34和整合素αV共染色，而与GFAP不共染色。此外，IL-13处理增加了VSMCs中tenascin-C的表达和分泌，并呈剂量依赖性。Tenascin-C暴露促进HRECs的增殖、迁移和管形成。Tenascin-C中和抗体明显阻断了暴露于vsmc - il -13条件培养基中的HRECs形成管。结论tenascin-C可从VSMCs分泌，促进PDR相关fvm血管生成。
Purpose We previously demonstrated that tenascin-C was highly expressed in the fibrovascular membranes (FVMs) of patients with proliferative diabetic retinopathy (PDR). However, its role in the pathogenesis of FVMs has not been determined. The purpose of this study was to investigate what role tenascin-C plays in the formation and angiogenesis of FVMs. Methods The level of tenascin-C was determined by sandwich enzyme-linked immunosorbent assay in the vitreous samples collected from patients with PDR and with a macular hole as control. The locations of tenascin-C, α- smooth muscle actin (SMA), CD34, glial fibrillary acidic protein (GFAP), and integrin αV in the FVMs from PDR patients were determined by immunohistochemistry. We also measured the in vitro expression of the mRNA and protein of tenascin-C in vascular smooth muscle cells (VSMCs) stimulated by interleukin (IL)-13. The effects of tenascin-C on cell proliferation, migration, and tube formation were determined in human retinal endothelial cells (HRECs) in culture. Results The mean vitreous levels of tenascin-C were significantly higher in patients with PDR than in patients with a macular hole (p<0.001). Double immunofluorescence analyses of FVMs from PDR patients showed that tenascin-C co-stained FVMs with α-SMA, CD34, and integrin αV but not with GFAP. In addition, IL-13 treatment increased both the expression and secretion of tenascin-C by VSMCs in a dose-dependent manner. Tenascin-C exposure promoted proliferation, migration, and tube formation in HRECs. Tenascin-C neutralizing antibody significantly blocked the tube formation by HRECs exposed to VSMC-IL-13-conditioned medium. Conclusions Our findings suggest that tenascin-C is secreted from VSMCs and promotes angiogenesis in the FVMs associated with PDR.