Chai Jiasui, Dong Jiejie, Hou Weixiao, Ning Jiayong, Liu Yan, Chen Yan, Zhang Tong, Guo Xingren, Yao Peng
Department of Hepatobiliary Surgery, Yuncheng Central Hospital Affiliated to Shanxi Medical University, No. 3690, Hedong East Street, Yanhu District, Yuncheng, 044000, Shanxi Province, China.
Dig Dis Sci. 2025 Jul 2. doi: 10.1007/s10620-025-09186-6.
Increasing evidence has revealed that aberrantly expressed miRNAs play important roles in stimulating pathogenesis and maintaining disease progression. However, the potential miRNAs involved in acute pancreatitis (AP) are still unclear. The present study aimed to explore the clinical significance of hsa-miR-4695-5p and its role behind AP progression.
Serum samples from 86 patients with AP (mild: 19 cases, moderate: 25 cases; severe: 42 cases) and 33 healthy cases were collected, and hsa-miR-4695-5p levels were detected by qRT-PCR analysis. Receiver operating characteristic (ROC) curves were employed to assess the diagnostic performance of hsa-miR-4695-5p. Caerulein-treated HPDE6-C7 cells were served as an in vitro cellular model for AP, cell viability, pro-inflammatory cytokines (TNF-α, IL-6 and IL-33) and oxidative stress were detected after silencing or overexpression of hsa-miR-4695-5p by CCK-8, ELISA, MDA and SOD kits. The influence of hsa-miR-4695-5p on macrophage polarization was verified through cell co-cultivation system by the detection of M1 and M2 macrophage markers. The targeting relationships between hsa-miR-4695-5p and sestrin 2 (SESN2) were confirmed by bioinformatics, luciferase reporter and RNA pull-down assays. In addition, a mouse AP model was used to confirm hsa-miR-4695-5p's role in AP.
We found that hsa-miR-4695-5p was upregulated in patients with AP relative to normal cases, and higher hsa-miR-4695-5p expression was observed in severe AP cases relative to mild and moderate AP cases. The combination of hsa-miR-4695-5p and serum amylase showed better diagnostic efficacy for severe AP than hsa-miR-4695-5p alone (AUC of 0.964 and 0.847). Interfering hsa-miR-4695-5p improved cell viability and increased SOD activity in caerulein-treated HPDE6-C7 cells, decreased MDA, TNF-α, IL-6 and IL-33 levels, and reduced the macrophage polarization toward M1. Reinforcing hsa-miR-4695-5p caused the opposite effects. SESN2 levels were downregulated in patients with severe AP and caerulein-treated HPDE6-C7 cells, and SESN2 was a direct target of hsa-miR-4695-5p. Moreover, SESN2 knockdown reversed the effects of interfering hsa-miR-4695-5p in caerulein-treated HPDE6-C7 cells. In vivo injection of hsa-miR-4695-5p antagomir attenuated pathological injury in AP mice.
Upregulation of serum hsa-miR-4695-5p could be a novel biomarker for diagnosis of severe AP, and hsa-miR-4695-5p may promote pathological injury in severe AP via targeting SESN2 expression.
越来越多的证据表明，异常表达的微小RNA（miRNA）在促进疾病发生和维持疾病进展中起重要作用。然而，参与急性胰腺炎（AP）的潜在miRNA仍不清楚。本研究旨在探讨hsa-miR-4695-5p的临床意义及其在AP进展中的作用机制。
收集86例AP患者（轻度：19例，中度：25例；重度：42例）和33例健康对照者的血清样本，采用qRT-PCR分析检测hsa-miR-4695-5p水平。采用受试者工作特征（ROC）曲线评估hsa-miR-4695-5p的诊断效能。用雨蛙肽处理的人胰腺导管上皮细胞（HPDE6-C7）作为AP的体外细胞模型，通过CCK-8、ELISA、丙二醛（MDA）和超氧化物歧化酶（SOD）试剂盒检测hsa-miR-4695-5p沉默或过表达后细胞活力、促炎细胞因子（肿瘤坏死因子-α、白细胞介素-6和白细胞介素-33）及氧化应激水平。通过细胞共培养系统检测M1和M2巨噬细胞标志物，验证hsa-miR-4695-5p对巨噬细胞极化的影响。通过生物信息学、荧光素酶报告基因和RNA下拉实验证实hsa-miR-4695-5p与 sestrin 2（SESN2）之间的靶向关系。此外，用小鼠AP模型验证hsa-miR-4695-5p在AP中的作用。
我们发现，与正常对照相比，AP患者血清中hsa-miR-4695-5p上调，且重度AP患者的hsa-miR-4695-5p表达高于轻度和中度AP患者。hsa-miR-4695-5p与血清淀粉酶联合检测对重度AP的诊断效能优于单独检测hsa-miR-4695-5p（曲线下面积分别为0.964和0.847）。干扰hsa-miR-4695-5p可提高雨蛙肽处理的HPDE6-C7细胞的活力，增加SOD活性，降低MDA、肿瘤坏死因子-α、白细胞介素-6和白细胞介素-33水平，并减少巨噬细胞向M1型极化。增强hsa-miR-4695-5p则产生相反的效果。重度AP患者和雨蛙肽处理的HPDE6-C7细胞中SESN2水平下调，且SESN2是hsa-miR-4695-5p的直接靶点。此外，敲低SESN2可逆转干扰hsa-miR-4695-5p对雨蛙肽处理的HPDE6-C7细胞的影响。体内注射hsa-miR-4695-5p拮抗剂可减轻AP小鼠的病理损伤。
血清hsa-miR-4695-5p上调可能是诊断重度AP的新型生物标志物，hsa-miR-4695-5p可能通过靶向SESN2表达促进重度AP的病理损伤。
