Roxvall L, Sennerby L, Johansson B R, Heideman M
Department of Surgery, Sahlgrens Hospital, Gothenburg, Sweden.
J Surg Res. 1990 Dec;49(6):504-13. doi: 10.1016/0022-4804(90)90175-2.
Trypsin-induced acute inflammation was studied in hamster cheek pouch using intravital microscopy, correlative histology, and electron microscopy. Vascular permeability changes were monitored with intravital fluoroscopy, after intravenous injection of FITC-dextran (Mw 150,000), by counting the number of FITC-dextran leakages around the vessels. The number of extravasated polymorphonuclear leukocytes (PMNLs) was calculated by a histological technique. A dose-dependent increase in the number of FITC-dextran leakages, as well as the number of accumulated PMNLs, was found when trypsin was locally deposited in concentrations of 0.25-2.5 microM (15 microliters during 5 min). Local deposition of autologous serum treated with trypsin at final concentrations of 0.25-2.5 microM caused an increase in vascular permeability as equally pronounced as that of pure trypsin, but only a moderate PMNL accumulation which was not dose dependent. Trypsin at a 25 microM concentration resulted in numerous microbleedings and cessation of flow. The electron microscopy demonstrated inflammatory events (PMNL adhesion, diapedesis, and interstitial infiltration) in all treatment groups but they were more pronounced after trypsin exposure. Trypsin did not cause disintegration, cellular lysis, or increased mast cell degranulation. The permeability changes induced by trypsin (2.5 microM) and trypsinated serum (2.5 microM) were significantly suppressed by the addition of the chelating agent potassium-EDTA to the reaction mixture, indicating a calcium- or magnesium-dependent mechanism. Pretreatment of the animals with cobra venom factor (CVF), by which the plasma C3 concentration was reduced to less than 10%, inhibited the vascular leakages almost completely. The trypsin-induced accumulation of PMNLs was significantly reduced by potassium-EDTA as well as by pretreatment with CVF (P less than 0.01). These findings indicate a central role of complement activation in trypsin-induced acute inflammation in the hamster cheek pouch.
利用活体显微镜检查、相关组织学和电子显微镜技术，在仓鼠颊囊研究了胰蛋白酶诱导的急性炎症。静脉注射异硫氰酸荧光素标记的葡聚糖（分子量150,000）后，通过计数血管周围异硫氰酸荧光素标记的葡聚糖渗漏数量，采用活体荧光透视法监测血管通透性变化。采用组织学技术计算渗出的多形核白细胞（PMNL）数量。当胰蛋白酶以0.25 - 2.5微摩尔/升的浓度（5分钟内15微升）局部沉积时，发现异硫氰酸荧光素标记的葡聚糖渗漏数量以及积聚的PMNL数量呈剂量依赖性增加。用胰蛋白酶处理至终浓度为0.25 - 2.5微摩尔/升的自体血清局部沉积，导致血管通透性增加，其程度与纯胰蛋白酶相同，但仅引起中度的PMNL积聚，且不依赖剂量。25微摩尔/升浓度的胰蛋白酶导致大量微出血和血流停止。电子显微镜显示所有治疗组均有炎症事件（PMNL黏附、渗出和间质浸润），但胰蛋白酶暴露后更为明显。胰蛋白酶未引起细胞崩解、细胞溶解或肥大细胞脱颗粒增加。向反应混合物中添加螯合剂乙二胺四乙酸钾可显著抑制胰蛋白酶（2.5微摩尔/升）和经胰蛋白酶处理的血清（2.5微摩尔/升）诱导的通透性变化，表明存在钙或镁依赖性机制。用眼镜蛇毒因子（CVF）预处理动物，使血浆C3浓度降至10%以下，几乎完全抑制了血管渗漏。乙二胺四乙酸钾以及用CVF预处理均显著降低了胰蛋白酶诱导的PMNL积聚（P < 0.01）。这些发现表明补体激活在仓鼠颊囊胰蛋白酶诱导的急性炎症中起核心作用。
