This research aims to investigate the effect of gemcitabine (GEM) on various activities and functions of macrophages. Phagocytosis, cell autophagy and reactive oxygen species (ROS) were analysed by laser scanning confocal microscope. The cell cycle status and major histocompatibility complex II (MHC-II) expression were examined by flow cytometry. Inflammatory cytokine secretion such as tumour necrosis factor α (TNF-α) and interleukin 6 (IL-6) was detected by Elisa assay. The expression of proteins was analysed by western blot method. The results revealed that GEM-induced immune inhibition of M1-type RAW264.7 macrophages activated by interferon-γ (IFN-γ) and lipopolysaccharide (LPS). We also found that GEM inhibited autophagy, as evidenced by the reduced formation of autophagosome-like vacuoles and autophagosomes. Further study showed that incubation of activated macrophages with the autophagy inhibitor 3-MA induced immune suppression. In contrast, treatment with the autophagy inducer trehalose (Tre) restored phagocytosis, TNF-α and IL-6 secretion, and MHC-II expression in GEM-induced immune-inhibited macrophages. GEM reduced immune effect of M1-type RAW264.7 macrophages via inhibiting TNF-α, IL-6 and MHC-II expression. Furthermore, activation of autophagy by Tre reversed GEM-induced immune inhibition of RAW264.7 macrophages.

本研究旨在探讨吉西他滨 (GEM) 对巨噬细胞各种活动和功能的影响。通过激光扫描共聚焦显微镜分析吞噬作用、细胞自噬和活性氧（ROS）。通过流式细胞术检查细胞周期状态和主要组织相容性复合物 II (MHC-II) 表达。Elisa法检测肿瘤坏死因子α（TNF-α）和白细胞介素6（IL-6）等炎症细胞因子的分泌。通过蛋白质印迹法分析蛋白质的表达。结果表明，GEM 诱导的对由干扰素-γ (IFN-γ) 和脂多糖 (LPS) 激活的 M1 型 RAW264.7 巨噬细胞的免疫抑制作用。我们还发现 GEM 抑制自噬，这可以通过减少自噬体样空泡和自噬体的形成来证明。进一步的研究表明，活化的巨噬细胞与自噬抑制剂 3-MA 一起孵育可诱导免疫抑制。相比之下，用自噬诱导剂海藻糖 (Tre) 治疗可恢复 GEM 诱导的免疫抑制巨噬细胞中的吞噬作用、TNF-α 和 IL-6 分泌以及 MHC-II 表达。GEM 通过抑制 TNF-α、IL-6 和 MHC-II 的表达来降低 M1 型 RAW264.7 巨噬细胞的免疫作用。此外，Tre 激活自噬可逆转 GEM 诱导的 RAW264.7 巨噬细胞的免疫抑制。7 巨噬细胞通过抑制 TNF-α、IL-6 和 MHC-II 的表达。此外，Tre 激活自噬可逆转 GEM 诱导的 RAW264.7 巨噬细胞的免疫抑制。7 巨噬细胞通过抑制 TNF-α、IL-6 和 MHC-II 的表达。此外，Tre 激活自噬可逆转 GEM 诱导的 RAW264.7 巨噬细胞的免疫抑制。