Dahlgren M E, Davies P, Bonney R J
Biochim Biophys Acta. 1980 Jul 3;630(3):338-51. doi: 10.1016/0304-4165(80)90282-2.
Cultured thioglycollate-elicited mouse peritoneal macrophages secrete an enzyme which hydrolyzes [3H]elastin prepared by NaB3H4 reduction of bovine ligamentum nuchae elastin. Over a 24 h culture period, 3.5 . 10(6) thioglycollate-elicited cells secrete sufficient enzyme to solubilize 200--350 micrograms [3H]elastin in 18h. Secretion at this rate continues for at least 6 days in culture. Secretion of the enzyme is stimulated 3-fold by exposure of the cultured cells to 10(-7) M phorbol myristate acetate, whereas the parent alcohol 4 alpha-phorbol is inactive in this respect. Enzyme activity is linearly related to the amount of conditioned medium assayed and is linear over incubation times up to 30 h. Unlabelled elastin competitively inhibits the solubilization of [3H]elastin. The solubilization rate is doubled if the substrate is pretreated with sodium dodecyl sulfate, but the rate of solubilization of this pretreated substrate increases with time. Resident peritoneal macrophages secrete barely detectable amounts of elastase, but phorbol myristate acetate (10(-7) M) stimulates its secretion in amounts comparable to those secreted by phorbol myristate acetate-stimulated thioglycollate-elicited cells. Dexamethasone (10(-9) M) inhibits phorbol myristate acetate-induced secretion by 50%, but 10(-6) M indomethacin is without effect. The secreted enzyme has the characteristics of a metalloproteinase.
培养的经巯基乙酸盐诱导的小鼠腹腔巨噬细胞分泌一种酶，该酶可水解通过用硼氢化钠还原牛项韧带弹性蛋白制备的[³H]弹性蛋白。在24小时的培养期内，3.5×10⁶个经巯基乙酸盐诱导的细胞分泌的酶足以在18小时内溶解200 - 350微克的[³H]弹性蛋白。以这种速率分泌至少持续6天。将培养的细胞暴露于10⁻⁷M佛波醇肉豆蔻酸酯乙酸酯可使该酶的分泌增加3倍，而母体醇4α - 佛波醇在这方面无活性。酶活性与所检测的条件培养基的量呈线性相关，并且在长达30小时的孵育时间内呈线性。未标记的弹性蛋白竞争性抑制[³H]弹性蛋白的溶解。如果底物用十二烷基硫酸钠预处理，溶解速率会加倍，但这种预处理底物的溶解速率会随时间增加。驻留腹腔巨噬细胞分泌的弹性蛋白酶量几乎检测不到，但佛波醇肉豆蔻酸酯乙酸酯（10⁻⁷M）刺激其分泌的量与经佛波醇肉豆蔻酸酯乙酸酯刺激的巯基乙酸盐诱导细胞分泌的量相当。地塞米松（10⁻⁹M）可抑制佛波醇肉豆蔻酸酯乙酸酯诱导的分泌50%，但10⁻⁶M吲哚美辛无作用。分泌的酶具有金属蛋白酶的特性。
