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首页> 外文期刊>Otolaryngology--head and neck surgery: official journal of American Academy of Otolaryngology-Head and Neck Surgery >Analysis of protease activity in human otitis media.
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Analysis of protease activity in human otitis media.

机译：分析人类中耳炎中蛋白酶的活性。

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摘要

Chronic otitis media is a common problem associated with a nonintact tympanic membrane frequently involving Staphylococcus aureus and Pseudomonas aeruginosa. The virulence of Pseudomonas bacteria is related to the production of two matrix metalloproteinases, elastase and alkaline protease. Serine proteases, such as neutrophil elastase, are produced by the host inflammatory response. These proteases are thought to contribute to tissue destruction and assist bacterial invasion during infection. This preliminary study was done to identify protease activity in otorrhea samples from patients with otitis media and a nonintact tympanic membrane and to examine the ability of selective protease inhibitors to decrease protease activity. Ilomostat (galardin) is a synthetic, specific inhibitor of matrix metalloproteinases including P. aeruginosa elastase and alkaline protease, whereas alpha1-antitrypsin inhibits serine proteases including neutrophil elastase. Samples were collected and cultured from 20 patients with otorrhea resulting from tympanic membrane perforations or pressure-equalization tubes. A protease assay that used azocasein as the substrate was used to quantify protease activity, with and without addition of selective protease inhibitors. Cultures revealed P. aeruginosa alone in 7 samples, P. aeruginosa plus other organisms in 10, and S. aureus alone in 3. Protease activity was detected in 15 (75%) of the samples. A statistically significant (p < 0.05) decrease in protease activity was seen with the addition of alpha1-antitrypsin or Ilomostat plus alpha1-antitrypsin, but not with Ilomostat alone. Analyzing the 10 samples with the highest protease activity, a statistically significant decrease in activity was seen with Ilomostat or alpha1-antitrypsin alone and with both Ilomostat and alpha1-antitrypsin together. Bacteriologic type, source of sample, age and gender of the subject, and duration of infection were not significantly related to protease activity. This is the first study to quantify protease activity and inhibition by selective protease inhibitors in human otorrhea. Protease inhibitors effectively decrease protease activity in most cases and in addition to standard antibiotic therapy might prove beneficial in the treatment of otitis media with a nonintact tympanic membrane. This study supports future clinical investigations into the role of proteases and inhibition of protease activity in the treatment of otitis media.
机译：慢性中耳炎是与鼓膜不完整相关的常见问题，经常涉及金黄色葡萄球菌和铜绿假单胞菌。假单胞菌细菌的毒性与两种基质金属蛋白酶，弹性蛋白酶和碱性蛋白酶的产生有关。丝氨酸蛋白酶，例如嗜中性粒细胞弹性蛋白酶，是由宿主的炎症反应产生的。这些蛋白酶被认为有助于组织破坏并在感染过程中帮助细菌入侵。进行了这项初步研究，以鉴定中耳炎和鼓膜不完整患者的耳漏样品中的蛋白酶活性，并研究选择性蛋白酶抑制剂降低蛋白酶活性的能力。伊洛司他（加拉丁）是一种合成的，特异性的基质金属蛋白酶抑制剂，包括铜绿假单胞菌弹性蛋白酶和碱性蛋白酶，而α1-抗胰蛋白酶抑制丝氨酸蛋白酶，包括嗜中性粒细胞弹性蛋白酶。从鼓膜穿孔或压力平衡管引起的20例耳漏患者中收集样品并进行培养。使用偶氮酪蛋白作为底物的蛋白酶测定用于定量蛋白酶活性，添加和不添加选择性蛋白酶抑制剂。培养显示7个样品中只有铜绿假单胞菌，10个样品中只有铜绿假单胞菌和其他生物，而3个样品中只有金黄色葡萄球菌。在15个样品（75％）中检测到蛋白酶活性。加入α1-抗胰蛋白酶或伊洛司他加上α1-抗胰蛋白酶后，发现蛋白酶活性有统计学上显着的下降（p <0.05），但单独使用伊洛伐他汀则没有。分析10个具有最高蛋白酶活性的样品，单独使用Ilomostat或alpha1-antitrypsin以及同时使用Ilomostat和alpha1-antitrypsin时，发现活性有统计学上的显着下降。细菌学类型，样品来源，受试者的年龄和性别以及感染持续时间与蛋白酶活性没有显着相关。这是第一个量化蛋白酶活性和人类耳漏中选择性蛋白酶抑制剂抑制作用的研究。在大多数情况下，蛋白酶抑制剂可有效降低蛋白酶的活性，除标准抗生素治疗外，蛋白酶抑制剂还可证明对治疗鼓膜完整的中耳炎有益。这项研究支持对蛋白酶在中耳炎治疗中的作用和蛋白酶活性抑制的未来临床研究。

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