Department of Hepatopancreatobiliary Surgery, The Second People's Hospital of Quzhou, Quzhou, Zhejiang, China.
Department of Clinical Laboratory, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China.
PeerJ. 2023 Jun 30;11:e15612. doi: 10.7717/peerj.15612. eCollection 2023.
With the number of patients with acute pancreatitis (AP) increasing year by year, it is pressing to explore new key genes and markers for the treatment of AP. miR-455-3p/solute carrier family 2 member 1 (Slc2a1) obtained through bioinformatics analysis may participate in the progression of AP.
The C57BL/6 mouse model of AP was constructed for subsequent studies. Through bioinformatics analysis, the differentially expressed genes related to AP were screened and hub genes were identified. A caerulein-induced AP animal model was constructed to detect the pathological changes of mouse pancreas by HE staining. The concentrations of amylase and lipase were measured. Primary mouse pancreatic acinar cells were isolated and subjected to microscopy to observe their morphology. The enzymatic activities of trypsin and amylase were detected. The secretion of inflammatory cytokines in mouse were measured with the ELISA kits of TNF-, IL-6 and IL-1 to determine pancreatic acinar cell damage. A binding site between the Slc2a1 3' UTR region and the miR-455-3p sequence was verified by dual-luciferase reporter assay. The expression of miR-455-3p was quantified by qRT-PCR, and Slc2a1 were detected by western blot.
A total of five (Fyn, Gadd45a, Sdc1, Slc2a1, and Src) were identified by bioinformatics analysis, and miR-455-3p/Slc2a1 were further studied. HE staining results showed that the AP models were successfully established by caerulein induction. In mice with AP, the expression of miR-455-3p was reduced, while that of Slc2a1 was increased. In the caerulein-induced cell model, the expression of Slc2a1 was significantly reduced after intervention of miR-455-3p mimics, whereas increased after miR-455-3p inhibitor treatment. miR-455-3p decreased the secretion of inflammatory cytokines in the cell supernatant, reduced the activity of trypsin and amylase, and alleviated the cell damage induced by caerulein. In addition, Slc2a1 3'UTR region was bound by miR-455-3p, and its protein expression was also regulated.
miR-455-3p alleviated caerulein-induced mouse pancreatic acinar cell damage by regulating the expression of Slc2a1.
随着急性胰腺炎（AP）患者数量逐年增加，迫切需要探索 AP 治疗的新的关键基因和标志物。通过生物信息学分析获得的 miR-455-3p/溶质载体家族 2 成员 1（Slc2a1）可能参与 AP 的进展。
构建 C57BL/6 小鼠 AP 模型进行后续研究。通过生物信息学分析筛选出与 AP 相关的差异表达基因，并鉴定出核心基因。通过胆胰管内注射蛙皮素（cerulein）建立 AP 动物模型，通过 HE 染色观察小鼠胰腺的病理变化。检测血清淀粉酶和脂肪酶的浓度。分离原代小鼠胰腺腺泡细胞，通过显微镜观察其形态。检测胰酶和淀粉酶的酶活性。用 TNF-、IL-6 和 IL-1 的 ELISA 试剂盒测量小鼠的炎症细胞因子分泌，以确定胰腺腺泡细胞的损伤情况。通过双荧光素酶报告实验验证 Slc2a1 3'UTR 区域与 miR-455-3p 序列的结合位点。通过 qRT-PCR 定量检测 miR-455-3p 的表达，通过 Western blot 检测 Slc2a1 的表达。
通过生物信息学分析共鉴定出 5 个（Fyn、Gadd45a、Sdc1、Slc2a1 和 Src）核心基因，进一步研究了 miR-455-3p/Slc2a1。HE 染色结果表明，通过胆胰管内注射蛙皮素成功建立了 AP 模型。在 AP 小鼠中，miR-455-3p 的表达降低，而 Slc2a1 的表达升高。在胆胰管内注射蛙皮素诱导的细胞模型中，miR-455-3p 模拟物干预后 Slc2a1 的表达明显降低，而 miR-455-3p 抑制剂处理后则升高。miR-455-3p 降低了细胞上清液中炎症细胞因子的分泌，降低了胰酶和淀粉酶的活性，并减轻了蛙皮素诱导的细胞损伤。此外，Slc2a1 3'UTR 区域与 miR-455-3p 结合，其蛋白表达也受到调节。
miR-455-3p 通过调节 Slc2a1 的表达缓解了胆胰管内注射蛙皮素诱导的小鼠胰腺腺泡细胞损伤。
