Rotatori D S, Kerr N C, Raphael B, McLaughlin B J, Shimizu R, Stern G A, Schultz G S
Department of Surgery, University of Florida College of Medicine, Gainesville.
Invest Ophthalmol Vis Sci. 1994 Jan;35(1):143-9.
To determine if a scrape injury to cat corneal endothelial cells increases the level of mitogenic proteins such as transforming growth factor alpha (TGF alpha) in aqueous humor.
Aqueous humor of cats was collected at 0, 2, 6, and 24 hours after wounding the endothelium by contact with a cannula tip. Aqueous humor samples collected from sham-wounded cats served as controls. Aqueous humor samples were analyzed for levels of protein, for mitogenic activity using incorporation of tritiated thymidine by cultures of bovine corneal endothelial cells, and for immunoreactive TGF alpha protein using a specific radioimmunoassay.
The average protein level in aqueous humor obtained before wounding was low (0.5 mg/ml), increased 26-fold at 2 hours after injury (13 mg/ml), then progressively decreased at 6 hours (8 mg/ml) and 24 hours (2 mg/ml). Levels of mitogenic activity of aqueous humor samples collected 2, 6, and 24 hours after wounding were 2-fold, 2.5-fold, and 0.6-fold higher, respectively, compared to the level of mitogenic activity measured in aqueous humor collected before wounding (0 hours) or in aqueous humor collected from sham-wounded eyes. TGF alpha concentration in aqueous humor collected before endothelial wounding was low (6.8 ng/ml), increased 14-fold 2 hours after wounding (97.4 ng/ml), then progressively decreased at 6 hours (63.3 ng/ml) and 24 hours (35.5 ng/ml) after wounding. TGF alpha concentrations in aqueous humor collected from sham-wounded eyes at 2 hours (9.5 ng/ml) and 6 hours (5.3 ng/ml) were not significantly different from prewound levels. Detergent extracts of bovine corneal endothelial cells contained substantial levels of TGF alpha immunoreactive protein (20 ng/mg protein).
Wounding of cat endothelium causes a rapid increase in mitogenic proteins in aqueous humor including TGF alpha, which may act by an autocrine mechanism to stimulate endothelial wound healing.
确定猫角膜内皮细胞的刮伤是否会增加房水中促有丝分裂蛋白的水平，如转化生长因子α（TGFα）。
通过套管尖端接触损伤内皮后，在0、2、6和24小时收集猫的房水。从假损伤猫收集的房水样本作为对照。分析房水样本中的蛋白质水平，使用牛角膜内皮细胞培养物掺入氚化胸腺嘧啶来检测促有丝分裂活性，并使用特异性放射免疫测定法检测免疫反应性TGFα蛋白。
受伤前获得的房水中平均蛋白质水平较低（0.5mg/ml），受伤后2小时增加26倍（13mg/ml），然后在6小时（8mg/ml）和24小时（2mg/ml）逐渐下降。受伤后2、6和24小时收集的房水样本的促有丝分裂活性水平分别比受伤前（0小时）或假损伤眼收集的房水中测得的促有丝分裂活性水平高2倍、2.5倍和0.6倍。内皮损伤前收集的房水中TGFα浓度较低（6.8ng/ml），受伤后2小时增加14倍（97.4ng/ml），然后在受伤后6小时（63.3ng/ml）和24小时（35.5ng/ml）逐渐下降。假损伤眼在2小时（9.5ng/ml）和6小时（5.3ng/ml）收集的房水中TGFα浓度与伤前水平无显著差异。牛角膜内皮细胞的去污剂提取物含有大量的TGFα免疫反应性蛋白（20ng/mg蛋白）。
猫内皮损伤导致房水中促有丝分裂蛋白迅速增加，包括TGFα，其可能通过自分泌机制刺激内皮伤口愈合。
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