Sahay Prity, Rao Aparna, Padhy Debananda, Sarangi Sarada, Das Gopinath, Reddy Mamatha M, Modak Rahul
Hyderabad Eye Research Foundation (HERF), LV Prasad Eye Institute, Patia, Bhubaneswar, Odisha, India 2KIIT School of Biotechnology, Patia, Bhubaneswar, Odisha, India.
Hyderabad Eye Research Foundation (HERF), LV Prasad Eye Institute, Patia, Bhubaneswar, Odisha, India.
Invest Ophthalmol Vis Sci. 2017 May 1;58(6):BIO106-BIO113. doi: 10.1167/iovs.17-21723.
To evaluate the differential expression of tear matrix metalloproteinases (MMP) 2 and 9 in of patients with various forms of glaucoma.
Tear samples were collected with a Schirmer's strip from 148 eyes of 113 patients (medically naïve patients with primary open-angle [POAG] or angle closure glaucoma [PACG] and those with pseudoexfoliation syndrome [PXF] or glaucoma [PXG]). These were compared to patients undergoing cataract surgery (controls) for this cross-sectional study. Functional activities of tear MMP-9 and MMP-2 were analyzed by gelatin zymography. Tenon's capsules (n = 15) were harvested from the inferior quadrant in those undergoing cataract surgery and protein expression of MMP-9 was analyzed by immunohistochemistry (IHC). Hydrogen peroxide (H2O2) stress-induced effects on in vitro activities of MMP-9 in human trabecular meshwork (HTM) cells were analyzed.
The MMP-9 activity in tears was increased significantly in POAG, (n = 27), PACG (n = 24), and PXF (n = 40) eyes compared to controls (n = 35), and was increased significantly in eyes with glaucoma compared to moderate/severe glaucoma (P < 0.001). The MMP-9 expression was significantly lower in PXG (n = 22) eyes. Immunohistochemistry of Tenon's capsule revealed increased expression of MMP-9 in primary glaucoma eyes. Increased MMP-9 activity was seen in in vitro by gelatin zymography and was confirmed by Western and immunofluorescent assay on HTM upon 800 and 1000 μM H2O2-induced stress for 2 to 3 hours with approximately 80% cell death.
Increased tear MMP-9 activity in early glaucoma and pseudoexfoliation syndrome suggesting activation of extracellular matrix (ECM) degradation can be used as a tear-based predictive biomarker. Decreased expression in advanced stages suggests exhaustion of the degradation response.
评估不同类型青光眼患者泪液中基质金属蛋白酶（MMP）2和9的差异表达。
采用泪液试纸从113例患者的148只眼中收集泪液样本（未经治疗的原发性开角型青光眼[POAG]或闭角型青光眼[PACG]患者以及假性剥脱综合征[PXF]或剥脱性青光眼[PXG]患者）。在这项横断面研究中，将这些患者与接受白内障手术的患者（对照组）进行比较。通过明胶酶谱法分析泪液MMP - 9和MMP - 2的功能活性。从接受白内障手术患者的下象限采集15个眼球筋膜囊，通过免疫组织化学（IHC）分析MMP - 9的蛋白表达。分析过氧化氢（H2O2）应激对人小梁网（HTM）细胞中MMP - 9体外活性的影响。
与对照组（n = 35）相比，POAG组（n = 27）、PACG组（n = 二十四）和PXF组（n = 40）眼中泪液中的MMP - 9活性显著增加，与中度/重度青光眼相比，青光眼患者眼中的MMP - 9活性显著增加（P < 0.001）。PXG组（n = 22）眼中的MMP - 9表达显著降低。眼球筋膜囊的免疫组织化学显示原发性青光眼眼中MMP - 9表达增加。通过明胶酶谱法在体外观察到MMP - 9活性增加，并通过Western和免疫荧光分析在800和1000μM H2O2诱导应激2至3小时后对HTM进行确认，此时细胞死亡率约为80%。
早期青光眼和假性剥脱综合征中泪液MMP - 9活性增加，提示细胞外基质（ECM）降解激活，可作为基于泪液的预测生物标志物。晚期表达降低提示降解反应耗竭。
