Department of Critical Care Medicine, Yijishan Hospital, The First Affiliated Hospital of Wannan Medical College, Wuhu, Anhui, China.
Department of Burn and Plastic Surgery, Yijishan Hospital, The First Affiliated Hospital of Wannan Medical College, Wuhu, Anhui, China.
Cell Biol Int. 2021 Sep;45(9):1935-1944. doi: 10.1002/cbin.11633. Epub 2021 Jun 15.
Intestinal barrier dysfunction often occurs in various acute or chronic pathological conditions and has been identified as an important clinical problem. Herein, we explored the biological role and molecular mechanism of Polo-like kinase 1 (PLK1) and differentiation antagonizing non-protein coding RNA (DANCR) in intestinal barrier dysfunction caused by sepsis. RT-qPCR analysis was used to examine PLK1, miR-1306-5p, and DANCR expression in NCM460 cells after LPS treatment. TUNEL assay and Western blot analysis were performed to explore PLK1 function in cell apoptosis and intestinal barrier in vitro. Hematoxylin and eosin staining, Western blot analysis, and TUNEL assay were used to investigate DANCR function in the intestinal barrier and cell apoptosis in vivo. The interaction between miR-1306-5p and PLK1 (or DANCR) was validated by luciferase reporter assay. As a result, PLK1 overexpression decreased cell apoptosis and promoted intestinal barrier function. Moreover, DANCR was validated as a sponge of miR-1306-5p to target PLK1. In addition, we found that DANCR overexpression decreased intestinal mucosal permeability and colon mucosa epithelial cell apoptosis in vivo. Conclusively, DANCR improved intestinal barrier dysfunction and alleviated epithelial injury by targeting the miR-1306-5p/PLK1 axis in sepsis.
肠道屏障功能障碍常发生于各种急性或慢性病理状态，已被确定为一个重要的临床问题。在此，我们探讨了丝氨酸/苏氨酸激酶 Polo 样激酶 1（PLK1）和差异表达反义非编码 RNA（DANCR）在脓毒症引起的肠道屏障功能障碍中的生物学作用和分子机制。用 LPS 处理 NCM460 细胞后，通过 RT-qPCR 分析检测 PLK1、miR-1306-5p 和 DANCR 的表达。通过 TUNEL assay 和 Western blot 分析，在体外研究 PLK1 对细胞凋亡和肠道屏障的功能。通过苏木精和伊红染色、Western blot 分析和 TUNEL assay ，研究体内 DANCR 在肠道屏障和细胞凋亡中的功能。通过荧光素酶报告实验验证 miR-1306-5p 与 PLK1（或 DANCR）之间的相互作用。结果表明，PLK1 的过表达降低了细胞凋亡，促进了肠道屏障功能。此外，DANCR 被证实为 miR-1306-5p 的海绵，可靶向 PLK1。此外，我们发现 DANCR 的过表达降低了脓毒症患者体内的肠黏膜通透性和结肠黏膜上皮细胞凋亡。总之，DANCR 通过靶向 miR-1306-5p/PLK1 轴改善了脓毒症中的肠道屏障功能障碍和上皮损伤。
