Trypsinogen activation in rat pancreatic acinar cells hyperstimulated by caerulein
Trypsinogen activation in rat pancreatic acinar cells hyperstimulated by caerulein
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DOI：
10.1016/s0925-4439(97)00082-3
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发表时间：
1997-12-31
影响因子：
6.2
通讯作者：
Lippert, H
中科院分区：
文献类型：
--
作者：
Halangk, W;Stürzebecher, J;Lippert, H
不适当的胰蛋白酶原激活被讨论为促分泌素诱导的急性胰腺炎模型中的早期细胞内事件。然而，胰蛋白酶原激活的机制尚未得到很好的表征。在目前的工作中，使用双-(CBZ-精氨酰基)-罗丹明 110 [(CBZ-Arg)(2)-Rho 110] 作为胰蛋白酶的细胞渗透底物，并独立地通过胰蛋白酶原激活肽 (TAP) 的形成，研究了完整腺泡细胞中的胰蛋白酶原激活。用 10nM 雨蛙素预孵育可将 Rho 110 底物裂解增加三倍以上。这种蛋白水解活性对苯甲脒 (BA) 型丝氨酸蛋白酶抑制剂完全敏感。酶活性的出现与 TAP 的形成同时发生。高分子大豆胰蛋白酶抑制剂缺乏作用表明细胞内底物裂解。组织蛋白酶 B 抑制剂 CA-074 既不阻止雨蛙素诱导的 TAP 形成，也不阻止 (CBZ-Arg)(2) Rho 110 裂解活性。 BA 抑制 Rho 110 底物裂解并显着减少 TAP 形成。这些结果表明，雨伞素过度刺激的腺泡细胞中胰蛋白酶原的激活可能独立于组织蛋白酶 B 的活性而发生。相反，BA 的作用表明丝氨酸蛋白酶在胰蛋白酶原激活中的作用。 (C) 1997 Elsevier Science B.V.
Inappropriate trypsinogen activation is discussed as an early intracellular event in the secretagogue-induced model of acute pancreatitis. However, the mechanisms by which trypsinogen is activated are not well characterized. In the present work, trypsinogen activation was studied in intact acinar cells using bis-(CBZ-arginyl)-Rhodamine 110 [(CBZ-Arg)(2)-Rho 110] as a cell-permeant substrate for trypsin and also independently via the formation of trypsinogen activation peptide (TAP). Preincubation with 10nM caerulein increased the Rho 110-substrate cleavage more than threefold. This proteolytic activity was fully sensitive to a benzamidine (BA)-type serine protease inhibitor. The appearance of enzymatic activity was paralleled by the formation of TAP. The lack of effect of the high-molecular soybean trypsin inhibitor indicates an intracellular substrate cleavage. The cathepsin B inhibitor CA-074 prevented neither the caerulein-induced formation of TAP nor the (CBZ-Arg)(2) Rho 110-cleaving activity. BA inhibited the Rho 110-substrate cleavage and significantly reduced the TAP formation. These results show that trypsinogen activation in caerulein-hyperstimulated acinar cells may occur independently of the activity of cathepsin B. On the contrary, the effect of BA suggests the role of a serine protease in trypsinogen activation. (C) 1997 Elsevier Science B.V.