Knepper Paul A, Mayanil Chandra S K, Goossens William, Wertz Robert D, Holgren Cory, Ritch Robert, Allingham R Rand
Laboratory for Oculo-Cerebrospinal Investigation, Division of Neurosurgery, Children's Memorial Medical Center, Northwestern University Medical School, Chicago, Illinois 60614, USA.
Invest Ophthalmol Vis Sci. 2002 Jan;43(1):133-9.
To determine whether the cell adhesion molecule CD44, the principal receptor of hyaluronan, is altered in the aqueous humor and the anterior segment of patients with primary open-angle glaucoma (POAG).
The trabecular meshwork (TM), iris, ciliary body, and sclera of POAG and age-matched control eyes preserved in ethanol were microdissected and subjected to 1% Triton X-100 solubilization at 4 degrees C. Western blot analysis was performed using monoclonal antibodies that recognize either CD44H (hematopoietic; extracellular domain) or CD44S (soluble ectodomain). The concentration of soluble CD44S in aqueous and microdissected tissues was measured by enzyme-linked immunosorbent assay (ELISA).
ELISA of soluble CD44S of aqueous from eyes of patients with POAG indicated that the concentration of soluble CD44S is increased in comparison with that of aqueous from normal eyes (P < 0.0003). Western blot analysis and densitometry of POAG iris and ciliary body revealed a statistically significant increase in the Triton X-100 extraction of CD44H. The predominant increases were in the 180-kDa (P < 0.001) and the 85-kDa (P < 0.001) forms. ELISA of soluble CD44S indicated that the concentration is statistically decreased in iris (P < 0.05), ciliary body (P < 0.001), and TM (P < 0.005) of POAG eyes.
Increased amounts of soluble CD44S in POAG aqueous and Triton X-100-solubilized CD44H characterized POAG in the iris and ciliary body. These soluble CD44 isoforms may influence the activity of the transmembrane CD44H by acting as inhibitors of CD44H and, thereby, adversely influence the cell survival of TM and retinal ganglion cells in POAG.
确定透明质酸的主要受体——细胞黏附分子CD44在原发性开角型青光眼(POAG)患者的房水和眼前节中是否发生改变。
将保存在乙醇中的POAG患者及年龄匹配的对照眼的小梁网(TM)、虹膜、睫状体和巩膜进行显微切割,并在4℃下用1% Triton X-100进行溶解。使用识别CD44H(造血型;细胞外结构域)或CD44S(可溶性胞外结构域)的单克隆抗体进行蛋白质印迹分析。通过酶联免疫吸附测定(ELISA)测量房水和显微切割组织中可溶性CD44S的浓度。
对POAG患者眼房水中可溶性CD44S的ELISA检测表明,与正常眼房水相比,可溶性CD44S的浓度升高(P < 0.0003)。对POAG虹膜和睫状体的蛋白质印迹分析和光密度测定显示,CD44H的Triton X-100提取物有统计学意义的增加。主要增加的是180 kDa(P < 0.001)和85 kDa(P < 0.001)形式。可溶性CD44S的ELISA检测表明,POAG眼的虹膜(P < 0.05)、睫状体(P < 0.001)和TM(P < 0.005)中该浓度有统计学意义的降低。
POAG房水中可溶性CD44S的量增加以及Triton X-100溶解的CD44H是POAG虹膜和睫状体的特征。这些可溶性CD44异构体可能通过作为CD44H的抑制剂影响跨膜CD44H的活性,从而对POAG中小梁网和视网膜神经节细胞的细胞存活产生不利影响。