Ishfaq Muhammad, Wu Zhiyong, Wang Jian, Li Rui, Chen Chunli, Li Jichang
Heilongjiang Key Laboratory for Animal Disease Control and Pharmaceutical Development, College of Veterinary Medicine, Northeast Agricultural University, 600 Changjiang Road, Xiangfang District, Harbin 150030, PR China; Huanggang Normal University, College of Computer Science, Huanggang 438000, PR China.
Heilongjiang Key Laboratory for Animal Disease Control and Pharmaceutical Development, College of Veterinary Medicine, Northeast Agricultural University, 600 Changjiang Road, Xiangfang District, Harbin 150030, PR China.
Int Immunopharmacol. 2021 Dec;101(Pt B):108250. doi: 10.1016/j.intimp.2021.108250. Epub 2021 Oct 14.
Baicalin is a well-known flavonoid compound, possess therapeutic potential against inflammatory diseases. Previous studies reported that Mycoplasma gallisepticum (MG) induced inflammatory response and immune dysregulation inside the host body. However, the underlying molecular mechanisms of baicalin against MG-infected chicken-like macrophages (HD11 cells) are still illusive. Oxidant status and total reactive oxygen species (ROS) were detected by ELISA assays and flow cytometry respectively. Mitochondrial membrane potential (ΔΨ) was evaluated by immunofluorescence microscopy. Transmission electron microscopy was used for ultrastructural analysis. The hallmarks of inflammation and autophagy were determined by western blotting. Oxidative stress and reactive oxygen species (ROS) were significantly enhanced in the MG-infected HD11 cells. MG infection caused disruption in the mitochondrial membrane potential (ΔΨ) compared to the control conditions. Meanwhile, baicalin treatment reduced MG-induced reactive oxygen species (ROS), oxidative stress and alleviated the disruption in ΔΨ. The activities of inflammatory markers were significantly enhanced in the MG-infected HD11 cells. Increased protein expressions of TLR-2-NF-κB pathway, NLRP3-inflammasome and autophagy-related proteins were detected in the MG-infected HD11 cells. Besides, baicalin treatment significantly reduced the protein expressions of TLR-2-NF-κB pathway and NLRP3 inflammasome. While, the autophagy-related proteins were significantly enhanced with baicalin treatment in a dose-dependent manner in the MG-infected HD11 cells. The results showed that baicalin prevented HD11 cells from MG-induced oxidative stress and inflammation via the opposite modulation of TLR-2-NF-κB-mediated NLRP3-inflammasome pathway and autophagy, and baicalin could be a promising candidate for the prevention of inflammatory effects caused by MG-infection in macrophages.
黄芩苷是一种著名的黄酮类化合物，具有治疗炎症性疾病的潜力。先前的研究报道，鸡毒支原体（MG）可诱导宿主体内的炎症反应和免疫失调。然而，黄芩苷对MG感染的鸡源巨噬细胞（HD11细胞）的潜在分子机制仍不清楚。分别通过ELISA检测和流式细胞术检测氧化状态和总活性氧（ROS）。通过免疫荧光显微镜评估线粒体膜电位（ΔΨ）。透射电子显微镜用于超微结构分析。通过蛋白质印迹法确定炎症和自噬的标志。在MG感染的HD11细胞中，氧化应激和活性氧（ROS）显著增强。与对照条件相比，MG感染导致线粒体膜电位（ΔΨ）破坏。同时，黄芩苷处理可降低MG诱导的活性氧（ROS）、氧化应激，并减轻ΔΨ的破坏。在MG感染的HD11细胞中，炎症标志物的活性显著增强。在MG感染的HD11细胞中检测到TLR-2-NF-κB途径、NLRP3炎性小体和自噬相关蛋白的蛋白表达增加。此外，黄芩苷处理显著降低了TLR-2-NF-κB途径和NLRP3炎性小体的蛋白表达。而在MG感染的HD11细胞中，黄芩苷处理以剂量依赖性方式显著增强了自噬相关蛋白。结果表明，黄芩苷通过对TLR-2-NF-κB介导的NLRP3炎性小体途径和自噬的反向调节，防止HD11细胞受到MG诱导的氧化应激和炎症，黄芩苷可能是预防MG感染巨噬细胞引起的炎症效应的有前途的候选药物。
