Abstract

Background

Severe acute pancreatitis (SAP) is a common acute abdominal disease with high morbidity and mortality. However, the mechanism underlying SAP is still unclear.

Methods

Cerulean and LPS (Cer-LPS) was used to establish a rat model and an in vitro model of SAP. qRT-PCR, western blot and IHC were determined to analyse the expression of mRNA and proteins. IL-1β, TNF-α and IL-6 levels were measured applying ELISA. H&E staining was determined to observe the pathological changes. Apoptosis was tested by AV-PI staining using flow cytometry. CCK8 assay was taken to detect cell viability. Cell migration was assessed by transwell assay. Tube formation assay was conducted to evaluate angiogenesis. Luciferase assay was used to detect relationship of miR-20b-5p and AKT3.

Results

MiR-20b-5p was lowly expressed in SAP models both in vivo and in vitro. Overexpression of miR-20b-5p restrained inflammation and apoptosis in Cer-LPS treated pancreatic acinar cells. Furthermore, miR-20b-5p promoted the angiogenesis of vascular endothelial cells, since the viability, migration and the capability of tube formation were increased by miR-20b-5p. Mechanically, miR-20b-5p directly targeted AKT3 to promote autophagy. Furthermore, miR-20b-5p could prevent the inflammation, apoptosis and enhance angiogenesis via enhancing autophagy, which was verified in vivo.

Conclusion

This study demonstrated miR-20b-5p attenuates SAP through directly targeting AKT3 to regulate autophagy, subsequently inhibit inflammation and apoptosis, and promote angiogenesis. Our findings suggested a novel target of miR-20b-5p for the therapy of SAP.

中文翻译：

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MiR-20b-5p 通过靶向 AKT3 的自噬调节重症急性胰腺炎的炎症、细胞凋亡和血管生成

摘要

背景

重症急性胰腺炎（SAP）是一种常见的急性腹部疾病，发病率和死亡率都很高。然而，SAP 的潜在机制仍不清楚。

方法

Cerulean and LPS（Cer-LPS）用于建立大鼠模型和SAP体外模型。测定qRT-PCR、蛋白质印迹和IHC以分析mRNA和蛋白质的表达。应用ELISA测量IL-1β、TNF-α和IL-6水平。测定H&E染色观察病理变化。使用流式细胞仪通过 AV-PI 染色检测细胞凋亡。采用CCK8测定法检测细胞活力。通过transwell测定评估细胞迁移。进行管形成测定以评估血管生成。荧光素酶测定用于检测miR-20b-5p与AKT3的关系。

结果

MiR-20b-5p在体内和体外的 SAP 模型中均低表达。miR-20b-5p 的过表达抑制了 Cer-LPS 处理的胰腺腺泡细胞的炎症和细胞凋亡。此外，miR-20b-5p 促进血管内皮细胞的血管生成，因为 miR-20b-5p 增加了活力、迁移和管形成能力。机械地，miR-20b-5p 直接靶向 AKT3 以促进自噬。此外，miR-20b-5p可以通过增强自噬来预防炎症、细胞凋亡和促进血管生成，这在体内得到了验证。

结论

本研究表明 miR-20b-5p 通过直接靶向 AKT3 来减弱 SAP，从而调节自噬，随后抑制炎症和细胞凋亡，并促进血管生成。我们的研究结果表明 miR-20b-5p 的新靶点可用于治疗 SAP。