LncRNA TCONS_00134892/miRNA-21/Trim33调控轴介导的激活的酶原颗粒泛素化在胰腺腺泡细胞选择性自噬中的作用研究

Recent studies have shown that a novel selective form of autophagy arised during acute pancreatitis (AP), which is named Zymophagy, a cellular process to specifically detect and degrade secretory granules containing activated enzymes before they can digest the organ. This process is mediated by ubiquitination modification. In our previous study, based on the bioinformatics and molecular biology experiment, we found that Trim33 (a E3 ubiquitin-protein ligase) is highly expressed during the process of Zymophagy, and lncRNA TCONS_00134892 could promote the expression of Trim33 by binding to miR-21, which in turn promoted the ubiquitination of the activated zymogen granules.In this study, we will confirm the competitive binding of lncRNA TCONS_00134892 and Trim33 to miR-21, and further establish the control relationship of lncRNA TCONS_00134892/miR-21/Trim33 and their regulation effect by using various research methods such as gene transfection, bioinformatics and molecular biology.This project aimed to further study the mechanism of Zymophagy in AP, find new targets for AP treatment, and provide evidence for the clinical transformation.

新近研究发现急性胰腺炎（AP）时胰腺腺泡细胞能通过自噬选择性地降解激活的酶原颗粒，这一现象称为Zymophagy。泛素化修饰介导了Zymophagy对激活的酶原颗粒的选择性。本课题组前期利用系统生物学及分子学实验方法研究发现一种E3泛素-蛋白连接酶Trim33，在Zymophagy激活时高表达；lncRNA TCONS_00134892可能通过竞争结合miR-21而促进Trim33表达，进而促进激活的酶原颗粒的泛素化修饰。本课题将在证实lncRNA TCONS_00134892能与Trim33竞争结合miR-21的基础上，在细胞、动物2个实验水平，借助基因转染技术和各种分子学实验方法，来证实lncRNA TCONS_00134892/miR-21/Trim33自上而下的调控关系及调控效应。本项目深入研究AP时Zymophagy的发生机制，希冀找到AP治疗的新的干预靶点。

急性胰腺炎早期，腺泡细胞内的胰蛋白酶原被激活，细胞通过酶噬清除胰蛋白酶，避免损伤。研究表明，酶噬的底物是泛素化胰酶，但胰酶泛素化的机制和酶噬的调控机制尚不完全清楚。我们的研究结果表明Trim33可以提高细胞活力，减少细胞坏死，降低胰蛋白酶原活化。Trim33是一种关键的E3连接酶，介导胰蛋白酶泛素化。结果表明，过表达Trim33可显著提高VMP1 mRNA和蛋白水平。然而，敲掉Trim33产生了相反的效果，这表明Trim33作为转录介质，通过调节VMP1的表达来影响酶噬。此外，我们还探讨了Trim33分子的转录调控机制。我们的研究表明，lncRNA TCONS_00021785可以竞争性结合miR-21-5p上调Trim33，从而启动酶自噬并减少酶原激活。