Hamanaka K, Toki N, Tsushima H, Takasugi S
Arch Dermatol Res. 1984;276(6):370-4. doi: 10.1007/BF00413357.
In the present study, attempts were made to clarify the role of an elastase-like enzyme in Arthus reaction by using rabbit-peritoneal polymorphonuclear leukocytes (PMNs). It was first confirmed that the injection of large amounts of PMNs with antibody can increase tissue damage in reversed Arthus reaction. The five various fractions, which were intact PMNs, whole homogenate, granule fraction, nuclei and cell debris, and membrane extract, were prepared from rabbit-peritoneal exudate. Each fraction was intracutaneously injected into the abdominal wall of rabbit. It was of particular interest that the tissue damage induced by membrane extract occurred earlier and was more severe than that caused by the others. Furthermore, membrane extract possessed remarkably higher caseinolytic and elastinolytic activities than the others. The membrane extract was partially purified by Sephadex G-100 column chromatography. The protein concentration of the elastase-like enzyme solution was 3.64 mg/ml, and its specific activity was 195 caseinolytic units and 0.45 elastinolytic units per milligram of protein. The cutaneous reaction by membrane extract was dependent on the concentration of this elastase-like enzyme and could be strongly inhibited by Elastatinal. From these results, the authors suggest that an elastase-like enzyme which originates in PMN granules is the key enzyme that promotes the severity of tissue damage in Arthus reaction.
在本研究中,我们试图通过使用兔腹膜多形核白细胞(PMN)来阐明一种类弹性蛋白酶在阿瑟斯反应中的作用。首先证实,注射大量带有抗体的PMN可增加被动阿瑟斯反应中的组织损伤。从兔腹膜渗出液中制备了五个不同的组分,分别是完整的PMN、全匀浆、颗粒组分、细胞核和细胞碎片以及膜提取物。将每个组分皮内注射到兔的腹壁。特别有趣的是,膜提取物诱导的组织损伤比其他组分更早出现且更严重。此外,膜提取物的酪蛋白分解和弹性蛋白分解活性明显高于其他组分。通过Sephadex G - 100柱色谱对膜提取物进行了部分纯化。类弹性蛋白酶溶液的蛋白质浓度为3.64 mg/ml,其比活性为每毫克蛋白质195个酪蛋白分解单位和0.45个弹性蛋白分解单位。膜提取物引起的皮肤反应取决于这种类弹性蛋白酶的浓度,并且可被弹性蛋白酶抑制剂强烈抑制。根据这些结果,作者认为源自PMN颗粒的类弹性蛋白酶是促进阿瑟斯反应中组织损伤严重程度的关键酶。