Early trypsin activation develops independently of autophagy in caerulein-induced pancreatitis in mice.
Early trypsin activation develops independently of autophagy in caerulein-induced pancreatitis in mice.
复制标题
DOI：
10.1007/s00018-019-03254-7
复制
发表时间：
2020-05
期刊：
影响因子：
--
通讯作者：
Mayerle J
中科院分区：
文献类型：
--
作者：
Malla SR;Krueger B;Wartmann T;Sendler M;Mahajan UM;Weiss FU;Thiel FG;De Boni C;Gorelick FS;Halangk W;Aghdassi AA;Reinheckel T;Gukovskaya AS;Lerch MM;Mayerle J
胰腺内胰蛋白酶原过早激活被广泛认为是急性胰腺炎的始发事件。先前的研究也表明分泌囊泡、内体、溶酶体或自噬体/自噬溶酶体是胰蛋白酶原激活的主要位点，细胞损伤性蛋白水解级联反应源于此。为了确定初始胰蛋白酶原激活的亚细胞区室，我们对转基因轻链 3 (LC3)-GFP 自噬报告小鼠中雨蛙素诱导的胰腺炎的前 12 小时进行了时间分辨率分析。胰腺内胰蛋白酶活性在 60 分钟内增加，血清淀粉酶在 2 小时内增加，但荧光自噬体仅在胰腺炎发生 4 小时后形成，同时蛋白质印迹上从胞浆 LC3-I 转变为膜 LC3-II。 60 分钟时，较重的亚细胞级分中的活化胰蛋白酶与组织蛋白酶 B 共同分布，但不与自噬标记物 LC3 或自噬蛋白 16 (ATG16) 共同分布。对来自 LC3-GFP 小鼠的原发性胰腺腺泡进行超最大雨蛙素刺激表明，在接触雨蛙素的前 15 分钟内，胰蛋白酶原激活与自噬溶酶体的形成无关。共定位研究（使用 GFP-LC3 自噬体与 Ile-Pro-Arg-AMC 胰蛋白酶活性以及溶酶体相关膜蛋白 2 [LAMP-2] 与胰蛋白酶原激活肽 [TAP] 的免疫金标记）表明仅在较晚的时间点自噬溶酶体中存在活性胰蛋白酶。总之，在雨蛙素诱导的胰腺炎的起始阶段，过早的蛋白酶激活独立于自噬溶酶体的形成和分泌途径产生的囊泡而发生。然而，自噬可能在这种疾病的后期调节整体细胞内胰蛋白酶活性。
Premature intrapancreatic trypsinogen activation is widely regarded as an initiating event for acute pancreatitis. Previous studies have alternatively implicated secretory vesicles, endosomes, lysosomes, or autophagosomes/autophagolysosomes as the primary site of trypsinogen activation, from which a cell-damaging proteolytic cascade originates. To identify the subcellular compartment of initial trypsinogen activation we performed a time-resolution analysis of the first 12 h of caerulein-induced pancreatitis in transgenic light chain 3 (LC3)-GFP autophagy reporter mice. Intrapancreatic trypsin activity increased within 60 min and serum amylase within 2 h, but fluorescent autophagosome formation only by 4 h of pancreatitis in parallel with a shift from cytosolic LC3-I to membranous LC3-II on Western blots. At 60 min, activated trypsin in heavier subcellular fractions was co-distributed with cathepsin B, but not with the autophagy markers LC3 or autophagy protein 16 (ATG16). Supramaximal caerulein stimulation of primary pancreatic acini derived from LC3-GFP mice revealed that trypsinogen activation is independent of autophagolysosome formation already during the first 15 min of exposure to caerulein. Co-localization studies (with GFP-LC3 autophagosomes versus Ile–Pro–Arg–AMC trypsin activity and immunogold-labelling of lysosomal-associated membrane protein 2 [LAMP-2] versus trypsinogen activation peptide [TAP]) indicated active trypsin in autophagolysosomes only at the later timepoints. In conclusion, during the initiating phase of caerulein-induced pancreatitis, premature protease activation develops independently of autophagolysosome formation and in vesicles arising from the secretory pathway. However, autophagy is likely to regulate overall intracellular trypsin activity during the later stages of this disease.
登录
查看更多内容
影响因子：
15.9
作者：
Halangk, W;Lerch, MM;Deussing, J
通讯作者：
Deussing, J
影响因子：
4.8
作者：
Aghdassi, Ali A.;John, Daniel S.;Lerch, Markus M.
通讯作者：
Lerch, Markus M.
DOI：
10.1152/ajpgi.00315.2001
发表时间：
2002-02-01
影响因子：
4.5
作者：
Halangk, W;Krüger, B;Lerch, MM
通讯作者：
Lerch, MM
DOI：
10.1113/jp275879
发表时间：
2018-07
期刊：
The Journal of physiology
影响因子：
--
作者：
Chvanov M;De Faveri F;Moore D;Sherwood MW;Awais M;Voronina S;Sutton R;Criddle DN;Haynes L;Tepikin AV
通讯作者：
Tepikin AV
影响因子：
4.8
作者：
Grasso, Daniel;Ropolo, Alejandro;Vaccaro, Maria I.
通讯作者：
Vaccaro, Maria I.