Han Chenyang, Sheng Yongjia, Wang Jin, Zhou Xiaohong, Li Wenyan, Zhang Caiqun, Wu Shasha, Yang Yi, Ruan Shuiliang
The Second Affiliated Hospital of Jiaxing University, 314001, China.
The Second Affiliated Hospital of Jiaxing University, 314001, China.
Int Immunopharmacol. 2023 Apr;117:109992. doi: 10.1016/j.intimp.2023.109992. Epub 2023 Mar 11.
This work aimed to investigate the role of long non-coding RNA (lncRNA) PCSK6-AS1 in inflammatory bowel disease (IBD). The levels of PCSK6-AS1 in human samples were detected, and its target protein HIPK2 was explored by protein mass spectrometry and ground select test (GST) method. Meanwhile, the HIPK2-STAT1 interaction relation was verified by pull-down assay. In the mouse model, Dextran Sulfate Sodium（DSS） was used to induce mouse colitis, then the effect of PCSK6-AS1 on mouse mucosal barrier was detected by immunohistochemical (IHC) staining, hematoxylin and eosin (H&E) staining, and the proportion of T-helper cells 1（Th1) cells was measured by flow cytometry (FCM). For in-vitro experiments, Th0 cells were used as the objects, and the effect of PCSK6-AS1 on Th1 differentiation was explored by FCM and enzyme-linked immunosorbent assay (ELISA). According to our results, the expression of PCSK6-AS1 in colitis tissues increased. PCSK6-AS1 interacted with HIPK2 to promote the expression of the latter, while HIPK2 promoted STAT1 phosphorylation to regulate Th1 differentiation. Th1 differentiation accelerated the mucosal barrier injury and aggravated the progression of colitis. In the Th0 model, PCSK6-AS1 promoted Th1 differentiation. In the animal model, PCSK6-AS1 enhanced Th1 differentiation in the tissues, decreased the tight junction (TJ) protein levels, and improved the mucosal barrier permeability. Suppressing PCSK6-AS1 and the HIPK2 inhibitor tBID decreased Th1 differentiation and tissue inflammation. According to our results, PCSK6-AS1 promotes Th1 cell differentiation via the HIPK2-STAT1 signaling, thus aggravating the chronic colitis-related mucosal barrier damage and tissue inflammation. PCSK6-AS1 has an important role in the occurrence and development of IBD.
本研究旨在探讨长链非编码RNA（lncRNA）PCSK6-AS1在炎症性肠病（IBD）中的作用。检测了人样本中PCSK6-AS1的水平，并通过蛋白质质谱分析和谷胱甘肽S-转移酶（GST）法探索其靶蛋白HIPK2。同时，通过下拉试验验证了HIPK2与STAT1的相互作用关系。在小鼠模型中，使用葡聚糖硫酸钠（DSS）诱导小鼠结肠炎，然后通过免疫组织化学（IHC）染色、苏木精和伊红（H&E）染色检测PCSK6-AS1对小鼠黏膜屏障的影响，并通过流式细胞术（FCM）测量辅助性T细胞1（Th1）的比例。在体外实验中，以Th0细胞为对象，通过FCM和酶联免疫吸附测定（ELISA）探索PCSK6-AS1对Th1分化的影响。根据我们的结果，PCSK6-AS1在结肠炎组织中的表达增加。PCSK6-AS1与HIPK2相互作用以促进后者的表达，而HIPK2促进STAT1磷酸化以调节Th1分化。Th1分化加速了黏膜屏障损伤并加重了结肠炎的进展。在Th0模型中，PCSK6-AS1促进Th1分化。在动物模型中，PCSK6-AS1增强了组织中的Th1分化，降低了紧密连接（TJ）蛋白水平，并提高了黏膜屏障通透性。抑制PCSK6-AS1和HIPK2抑制剂tBID可降低Th1分化和组织炎症。根据我们的结果，PCSK6-AS1通过HIPK2-STAT1信号通路促进Th1细胞分化，从而加重慢性结肠炎相关的黏膜屏障损伤和组织炎症。PCSK6-AS1在IBD的发生和发展中起重要作用。
