College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
Int J Biol Macromol. 2021 Dec 15;193(Pt B):1871-1884. doi: 10.1016/j.ijbiomac.2021.11.019. Epub 2021 Nov 10.
This study is to investigate the effect of microRNA-1338 (miR-1338) on the activity of Kupffer cells (KCs) and its mechanism regulated by ophiopogon polysaccharide liposome (OPL). KCs was treated with different OPL after transfected with miR-1338 mimic and miR-1338 inhibitor. The secretion of NO and iNOS, the expression of catalase (CAT) and peroxidase (POD), the phagocytic activity, the expression of CD14 and MHC II, the apoptosis and the secretion of ROS were measured. In addition, the expressions of key signal factors TLR4, IKKβ, MyD88 and NF-κB in NF-κB signaling pathway were measured by real-time PCR and Western blot (WB). The results showed that OPL could promote the secretion of iNOS, the expression of POD, the phagocytosis, the mRNA expression of TLR4, MyD88, IKKβ and NF-κB, the protein expression of TLR4 and NF-κB, and inhibit the cell apoptosis and ROS secretion after transfected with miR-1338 mimic. After transfected with miR-1338 inhibitor, OPL could promote the secretion of NO and iNOS, the expression of POD, cell migration, phagocytosis, and inhibit cell apoptosis. Meanwhile, the mRNA expression of TLR4, MyD88, IKKβ and NF-κB and the protein expression of TLR4, MyD88 and NF-κB were promoted. These results suggested that OPL could activate TLR4-NF-κB signaling pathway and thereby improve the activity of KCs by regulating miR-1338.
本研究旨在探讨 microRNA-1338（miR-1338）对枯否细胞（KCs）活性的影响及其被麦冬多糖脂质体（OPL）调控的机制。KCs 经 miR-1338 模拟物和 miR-1338 抑制剂转染后，用不同的 OPL 处理。测量 NO 和 iNOS 的分泌、CAT 和 POD 的表达、吞噬活性、CD14 和 MHC II 的表达、细胞凋亡和 ROS 的分泌。此外，通过实时 PCR 和 Western blot（WB）测量 NF-κB 信号通路中关键信号因子 TLR4、IKKβ、MyD88 和 NF-κB 的表达。结果表明，OPL 可促进 iNOS 的分泌、POD 的表达、吞噬作用、TLR4、MyD88、IKKβ和 NF-κB 的 mRNA 表达、TLR4 和 NF-κB 的蛋白表达，抑制 miR-1338 模拟物转染后的细胞凋亡和 ROS 分泌。转染 miR-1338 抑制剂后，OPL 可促进 NO 和 iNOS 的分泌、POD 的表达、细胞迁移、吞噬作用，并抑制细胞凋亡。同时，TLR4、MyD88、IKKβ和 NF-κB 的 mRNA 表达以及 TLR4、MyD88 和 NF-κB 的蛋白表达均得到促进。这些结果表明，OPL 可通过调节 miR-1338 激活 TLR4-NF-κB 信号通路，从而提高 KCs 的活性。
