circSMARCA5通过miR-181-5p/LAMP-2轴调控急性胰腺炎细胞炎症损伤的机制研究

Mechanism of circSMARCA5-mediated regulation of inflammatory injury in acute pancreatitis through the miR-181-5p/LAMP-2 axis

摘要:

背景与目的 急性胰腺炎（AP）是一种以胰腺腺泡细胞损伤和炎症反应为特征的消化系统急症。环状RNA（circRNA）在炎症相关疾病中的调控作用受到广泛关注，但circSMARCA5在AP中的作用及机制尚不明确。本研究探讨circSMARCA5对AP细胞损伤的影响及其与miR-181-5p/LAMP-2轴的调控关系。方法 采用雨蛙肽刺激AR42J细胞建立体外AP模型。采用qRT-PCR检测circSMARCA5、miR-181-5p及LAMP-2表达；MTT法检测细胞活力；ELISA检测TNF-α、IL-1β和IL-6水平；双荧光素酶报告基因实验验证circSMARCA5与miR-181-5p、miR-181-5p与LAMP-2之间的靶向关系；通过过表达circSMARCA5、miR-181-5p和LAMP-2进行功能回复实验。结果 雨蛙肽处理后AR42J细胞中circSMARCA5表达显著降低，而miR-181-5p表达明显升高（均P<0.05）。过表达circSMARCA5可显著提高细胞活力，降低TNF-α、IL-1β和IL-6水平（均P<0.05）。双荧光素酶报告基因实验表明，circSMARCA5可直接结合miR-181-5p，LAMP-2为miR-181-5p的靶基因。miR-181-5p过表达可抑制LAMP-2表达，降低细胞活力并促进炎症因子释放，而circSMARCA5或LAMP-2过表达均可部分逆转上述作用。结论 circSMARCA5在AP细胞模型中表达下调，其可通过竞争性吸附miR-181-5p上调LAMP-2表达，减轻炎症反应并改善细胞损伤。circSMARCA5/miR-181-5p/LAMP-2轴可能成为AP治疗的新型分子靶点。

Abstract:

Background and Aims Acute pancreatitis (AP) is an acute gastrointestinal emergency characterized by damage to pancreatic acinar cells and an inflammatory response. Circular RNAs (circRNAs) have been implicated in the regulation of various inflammatory disorders; however, the role of circSMARCA5 in AP remains unclear. This study aimed to investigate the function of circSMARCA5 and its underlying mechanism involving the miR-181-5p/LAMP-2 axis in AP.Methods An in vitro AP model was established by treating AR42J cells with cerulein. The expression levels of circSMARCA5, miR-181-5p, and LAMP-2 were determined by qRT-PCR. Cell viability was assessed using the MTT assay, and the levels of TNF-α, IL-1β, and IL-6 were measured by ELISA. Dual-luciferase reporter assays were performed to verify the interactions between circSMARCA5 and miR-181-5p as well as between miR-181-5p and LAMP-2. Gain-of-function and rescue experiments were conducted to evaluate their biological effects.Results CircSMARCA5 expression was significantly decreased, whereas miR-181-5p expression was markedly increased in cerulein-treated AR42J cells (all P<0.05). Overexpression of circSMARCA5 significantly enhanced cell viability and reduced the secretion of TNF-α, IL-1β, and IL-6 (all P<0.05). Dual-luciferase reporter assays confirmed that circSMARCA5 directly bound to miR-181-5p, while LAMP-2 was identified as a downstream target of miR-181-5p. Overexpression of miR-181-5p suppressed LAMP-2 expression, decreased cell viability, and aggravated inflammatory responses. These effects were partially reversed by circSMARCA5 or LAMP-2 overexpression.Conclusion CircSMARCA5 alleviates inflammatory injury in AP by acting as a competing endogenous RNA for miR-181-5p and thereby upregulating LAMP-2 expression. The circSMARCA5/miR-181-5p/LAMP-2 regulatory axis may represent a potential therapeutic target for AP.