Transgenic Expression of PRSS1R122H Sensitizes Mice to Pancreatitis
Transgenic Expression of PRSS1R122H Sensitizes Mice to Pancreatitis
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PRSS1R122 H转基因表达对小鼠胰腺炎的致敏作用
DOI：
10.1053/j.gastro.2019.08.01
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发表时间：
2020-03-01
期刊：
影响因子：
29.4
通讯作者：
Logsdon, Craig D.
中科院分区：
文献类型：
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作者：
Huang, Haojie;Swidnicka-Siergiejko, Agnieszka Katarzyna;Logsdon, Craig D.
背景与目的：胰蛋白酶原基因(PRSS1)突变可导致人类遗传性胰腺炎。然而，目前还不清楚PRSS1的突变形式如何促进疾病的发展。我们研究了突变形式的人PRSS1在小鼠体内的表达效果。方法：在全长胰腺弹性蛋白酶基因启动子的控制下，我们在胰腺腺泡细胞中特异性地表达了编码R122H的突变形式的PRSS1(R122H)。不表达这些转基因的小鼠作为对照。小鼠注射雨蛙素诱导急性胰腺炎，或注射脂多糖诱导慢性胰腺炎。其他组的小鼠被喂以乙醇或高脂饮食来诱发胰腺炎。收集胰腺组织，进行组织学、免疫印迹、实时聚合酶链式反应和免疫组织化学分析。测定胰腺组织匀浆中胰酶活性和胰凝乳酶活性。采集血液，测定血清淀粉酶活性。结果：表达PRSS1或PRSS1(R122H)转基因小鼠的胰腺有局灶性炎症；这些病变在表达PRSS1(R122H)的小鼠中更为突出。与对照组相比，表达PRSS1或PRSS1(R122H)的小鼠胰腺中热休克蛋白70和核因子(红系衍生2)样2的水平增加，而糜蛋白酶C的水平降低。注射雨蛙素后，PRSS1或PRSS1(R122H)的表达增加增加胰腺组织的局灶性损伤，并增加急性胰腺炎的严重程度。与表达PRSS1(R122H)的小鼠或对照组小鼠相比，注射脂多糖加剧了PRSS1(R122H)小鼠的炎症。与表达PRSS1(R122H)的小鼠相比，表达PRSS1(R122H)的小鼠在酒精喂养或高脂饮食后患上更严重的胰腺炎。与表达PRSS1(R122H)的小鼠相比，表达PRSS1(R122H)的小鼠的胰腺有更多的DNA损伤、细胞凋亡和胶原沉积，并增加了胰酶活性和炎性细胞的渗透。结论：与表达PRSS1的小鼠或对照小鼠相比，在小鼠中表达编码PRSS1(R122H)的转基因可以促进炎症反应并增加胰腺炎的严重程度。这些小鼠可作为人类遗传性胰腺炎的模型，并可用于研究内毒素、乙醇或高脂饮食诱导胰腺炎的机制。
BACKGROUND & AIMS: Mutations in the trypsinogen gene (PRSS1) cause human hereditary pancreatitis. However, it is not clear how mutant forms of PRSS1 contribute to disease development. We studied the effects of expressing mutant forms of human PRSS1 in mice. METHODS: We expressed forms of PRSS1 with and without the mutation encoding R122H (PRSS1(R122H)) specifically in pancreatic acinar cells under control of a full-length pancreatic elastase gene promoter. Mice that did not express these transgenes were used as controls. Mice were given injections of caerulein to induce acute pancreatitis or injections of lipopolysaccharide to induce chronic pancreatitis. Other groups of mice were fed ethanol or placed on a high-fat diet to induce pancreatitis. Pancreata were collected and analyzed by histology, immunoblots, real-time polymerase chain reaction, and immunohistochemistry. Trypsin enzymatic activity and chymotrypsin enzymatic activity were measured in pancreatic homogenates. Blood was collected and serum amylase activity was measured. RESULTS: Pancreata from mice expressing transgenes encoding PRSS1 or PRSS1(R122H) had focal areas of inflammation; these lesions were more prominent in mice that express PRSS1(R122H). Pancreata from mice that express PRSS1 or PRSS1(R122H) had increased levels of heat shock protein 70 and nuclear factor (erythroid-derived 2)-like 2, and reduced levels of chymotrypsin C compared with control mice. Increased expression of PRSS1 or PRSS1(R122H) increased focal damage in pancreatic tissues and increased the severity of acute pancreatitis after caerulein injection. Administration of lipopolysaccharide exacerbated inflammation in mice that express PRSS1(R122H) compared to mice that express PRSS1 or control mice. Mice that express PRSS1(R122H) developed more severe pancreatitis after ethanol feeding or a high-fat diet than mice that express PRSS1 or control mice. Pancreata from mice that express PRSS1(R122H) had more DNA damage, apoptosis, and collagen deposition and increased trypsin activity and infiltration by inflammatory cells than mice that express PRSS1 or control mice. CONCLUSIONS: Expression of a transgene encoding PRSS1(R122H) in mice promoted inflammation and increased the severity of pancreatitis compared with mice that express PRSS1 or control mice. These mice might be used as a model for human hereditary pancreatitis and can be studied to determine mechanisms of induction of pancreatitis by lipopolysaccharide, ethanol, or a high-fat diet.