Maruyama Yuko, Maruyama Kazuichi, Kato Yukinari, Kajiya Kentaro, Moritoh Satoru, Yamamoto Kotaro, Matsumoto Yuko, Sawane Mika, Kerjaschki Dontscho, Nakazawa Toru, Kinoshita Shigeru
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Tohoku University Graduate School of Medicine, Department of Ophthalmology, Sendai, Japan.
Invest Ophthalmol Vis Sci. 2014 Jul 1;55(8):4813-22. doi: 10.1167/iovs.13-13711.
Podoplanin has been shown to be a reliable marker of lymphatic endothelium, but its role in the lymphatic system has not been well investigated. The purpose of this study is to investigate the role of podoplanin in lymphangiogenesis and macrophage functions under inflammatory conditions.
Mouse corneal suture and ear section models were used to induce lymphangiogenesis and macrophage infiltration. Antilymphatic vessel endothelial hyaluronan-1 (Anti-LYVE-1) antibody was used to visualize lymphatic vessels. Thioglycollate-induced macrophages (mps) were collected and cultured with lipopolysaccharide (LPS), IFN-γ, and anti-mouse podoplanin antibody (PMab-1). Podoplanin, NF-κB, and mitogen-activated protein kinase (MAPK) pathway expression were detected by Western blot analysis. The TNF-α secretion was measured by ELISA.
Administration of PMab-1, reduced lymphangiogenesis in the corneal suture and ear wound healing models. Also, PMab-1 suppressed mps infiltration at the site of wound healing. Moreover, administration of PMab-1 led to a significant suppression of the rejection reaction in the corneal transplantation model. Our in vitro experiments showed that PMab-1 suppressed TNF-α secretion from mps under inflamed conditions, especially secretion caused by LPS stimulation. We confirmed the effect of PMab-1 on mps under inflamed conditions with a Western blot experiment, which clearly showed that the phosphorylation signal of the MAPK and NF-κB pathways was suppressed by PMab-1.
Podoplanin neutralization resulted in inhibition of lymphatic growth associated with corneal and ear wound healing as well as mps inflammation. These data suggest that podoplanin is a novel therapeutic target for suppressing lymphangiogenesis and inflammation.
血小板源性生长因子已被证明是淋巴管内皮细胞的可靠标志物，但其在淋巴系统中的作用尚未得到充分研究。本研究的目的是探讨血小板源性生长因子在炎症条件下对淋巴管生成和巨噬细胞功能的作用。
采用小鼠角膜缝线和耳部切片模型诱导淋巴管生成和巨噬细胞浸润。使用抗淋巴管内皮透明质酸-1（Anti-LYVE-1）抗体可视化淋巴管。收集经巯基乙酸诱导的巨噬细胞（mps），并与脂多糖（LPS）、干扰素-γ和抗小鼠血小板源性生长因子抗体（PMab-1）一起培养。通过蛋白质免疫印迹分析检测血小板源性生长因子、核因子-κB和丝裂原活化蛋白激酶（MAPK）途径的表达。通过酶联免疫吸附测定法测量肿瘤坏死因子-α的分泌。
给予PMab-1可减少角膜缝线和耳部伤口愈合模型中的淋巴管生成。此外，PMab-1抑制了伤口愈合部位的mps浸润。此外，给予PMab-1可显著抑制角膜移植模型中的排斥反应。我们的体外实验表明，PMab-1在炎症条件下抑制了mps分泌肿瘤坏死因子-α，尤其是由LPS刺激引起的分泌。我们通过蛋白质免疫印迹实验证实了PMab-1在炎症条件下对mps的作用，该实验清楚地表明，PMab-1抑制了MAPK和核因子-κB途径的磷酸化信号。
血小板源性生长因子中和导致与角膜和耳部伤口愈合以及mps炎症相关的淋巴生长受到抑制。这些数据表明，血小板源性生长因子是抑制淋巴管生成和炎症的新治疗靶点。
