Department of Gastroenterology, Yizheng Hospital of Nanjing Drum Tower Hospital Group, Yizheng 211900, Jiangsu, China.
Department of General Surgery, Yizheng Hospital of Nanjing Drum Tower Hospital Group, Yizheng 211900, Jiangsu, China.
Int Immunopharmacol. 2022 Oct;111:109123. doi: 10.1016/j.intimp.2022.109123. Epub 2022 Aug 10.
Our previous studies have shown that salidroside (Sal) exerted a protective effect in severe acute pancreatitis (SAP) via inhibiting the inflammatory response. However, the molecular mechanism has not been fully elucidated.
Using SAP rat model and miRNA microarray, the effect of Sal on miRNA expression profiling was determined and then validated their changes by quantitative Real-time PCR (qRT-PCR). Then, SAP cell model, enzyme-linked immunosorbent assay (ELISA) and Cell Counting Kit-8 (CCK-8) assay were used to explore the biological function of miR-217-5p in vitro. Bioinformatics analysis, luciferase reporter assay and miRNA pulldown assay were performed to investigate the underlying mechanism of miR-217-5p in the protection of Sal against SAP.
Compared with SAP group, 21 differentially expressed miRNAs were identified in SAP + Sal group. The target genes of these miRNAs were strongly associated with regulation of transcription, Axon guidance, Pathways in cancer and MAPK signaling pathway. Among these miRNAs, miR-217-5p was the most downregulated miRNA. Sal treatment alleviated cell injury and reduced the production of pro-inflammatory cytokines. Whereas overexpression of miR-217-5p reversed the effects of Sal. We identified YY1 associated factor 2 (YAF2) as a direct target gene of miR-217-5p and Sal treatment could upregulate YAF2 expression via targeting miR-217-5p. Furthermore, knockdown of YAF2 counteracted Sal-induced alleviation of cell injury and inflammation. Moreover, Sal could suppress the activation of p38 MAPK pathway by regulating miR-217-5p/YAF2 axis.
Our findings for the first time highlighted that Sal alleviated pancreatic injury and inhibited inflammation by regulating miR-217-5p/YAF2 axis, which might provide new therapeutic strategies for SAP treatment.
我们之前的研究表明，红景天苷（Sal）通过抑制炎症反应对重症急性胰腺炎（SAP）发挥保护作用。然而，其分子机制尚未完全阐明。
使用 SAP 大鼠模型和 miRNA 微阵列，确定 Sal 对 miRNA 表达谱的影响，并用实时定量 PCR（qRT-PCR）验证其变化。然后，使用 SAP 细胞模型、酶联免疫吸附测定（ELISA）和细胞计数试剂盒-8（CCK-8）检测 miR-217-5p 在体外的生物学功能。通过生物信息学分析、荧光素酶报告基因检测和 miRNA 下拉实验探讨 miR-217-5p 在 Sal 保护 SAP 中的潜在作用机制。
与 SAP 组相比，SAP+Sal 组有 21 个差异表达 miRNA。这些 miRNA 的靶基因与转录调控、轴突导向、癌症途径和 MAPK 信号通路强烈相关。在这些 miRNA 中，miR-217-5p 是下调最明显的 miRNA。Sal 治疗可减轻细胞损伤，减少促炎细胞因子的产生。而过表达 miR-217-5p 则逆转了 Sal 的作用。我们鉴定出 YY1 相关因子 2（YAF2）是 miR-217-5p 的直接靶基因，Sal 治疗可通过靶向 miR-217-5p 上调 YAF2 的表达。此外，沉默 YAF2 可拮抗 Sal 诱导的细胞损伤和炎症减轻。此外，Sal 可通过调节 miR-217-5p/YAF2 轴抑制 p38 MAPK 通路的激活。
本研究首次表明，Sal 通过调节 miR-217-5p/YAF2 轴减轻胰腺损伤并抑制炎症反应，为 SAP 的治疗提供了新的治疗策略。
