Non-coding small RNA expression analysis of rat pancreatic acinar cells(AR42J) treated with sodium taurocholate

Acute pancreatitis (AP) is a common digestive disorder with high morbidity and mortality. At present, the pathogenic mechanisms of AP remain unclear. Pancreatic acinar intracellular trypsinogen activation is considered to be an important cause of AP and is an important event in the early stages of AP. The activation of trypsinogen is a key factor for the pancreas to maintain normal function and that the abnormal activation of trypsinogen in pancreatic acinar cells is an initiating factor for the occurrence of AP. In the past decade, microRNA-related research results suggest that small non-coding RNAs play an important role in AP. Recently, endogenous transfer RNA-derived small RNA (tsRNA), a newly identified non-coding small RNA, is reported to be associated with multiple diseases. tsRNAs can be broadly classified into two main groups: tiRNAs (tRNA halves) and tRFs (tRNA-derived fragments). tiRNAs are produced by specific cleavage in the anticodon loop under various stress conditions (29-50 nucleotides). tRFs are separated to 4 subtypes by their sites of origin in pre-tRNA or mature tRNA, and generally shorter than tiRNAs (16-28 nucleotides). Similar to the function of microRNA, tsRNA can inhibit their functions by binding to target genes. However, the role of tsRNA in regulating AP pathogenesis has not been investigated. In this experiment, sodium taurocholate was used to treat the rat pancreatic acinar cell (AR42J) to establish the AP-related intracellular activation of trypsinogen model. Then we used RNA sequencing to identify the differentially expressed non-coding small RNAs including microRNA and tsRNA in the cell model.急性胰腺炎（Acute pancreatitis, AP）是一种发病率与死亡率均较高的常见消化系统疾病。目前，AP的致病机制尚未完全阐明。胰腺腺泡细胞内胰蛋白酶原激活被认为是AP发生的重要病因，亦是AP病程早期的关键事件。胰蛋白酶原的正常激活是胰腺维持正常生理功能的关键因素，而胰腺腺泡细胞中胰蛋白酶原的异常激活则是AP发生的起始因素。近十年来，微小RNA（microRNA）相关研究成果表明，小型非编码RNA在AP的发生发展中发挥着重要作用。近期，一类新鉴定的非编码小RNA——内源性转运RNA衍生小RNA（tsRNA）被报道与多种疾病密切相关。tsRNA可大致分为两大主要类别：tRNA应激诱导小RNA（tiRNAs，即tRNA半分子）与tRNA衍生片段（tRFs）。其中，tiRNAs是在各类应激条件下于反密码子环处特异性切割产生的，长度为29~50个核苷酸。tRFs则根据其在前体转运RNA或成熟转运RNA上的起源位点分为4个亚型，且整体长度短于tiRNAs，约为16~28个核苷酸。与微小RNA的功能相似，tsRNA可通过结合靶基因来抑制其表达。然而，目前尚未有研究探讨tsRNA在调控AP致病过程中的作用。本实验采用牛磺胆酸钠处理大鼠胰腺腺泡细胞（AR42J），构建AP相关的细胞内胰蛋白酶原激活模型。随后通过RNA测序（RNA sequencing）技术，鉴定该细胞模型中包括微小RNA与tsRNA在内的差异表达非编码小RNA。