School of Chinese Medicine, Guangdong Pharmaceutical University, Guangzhou, Guangzhou 510006, China.
School of Pharmacy (The Center for Drug Research and Development), Guangdong Pharmaceutical University, Guangzhou, Guangzhou 510006, China.
J Immunol Res. 2022 Oct 19;2022:5882136. doi: 10.1155/2022/5882136. eCollection 2022.
Targeting dendritic cells (DCs) metabolism-related pathways and in-situ activation of DCs have become a new trend in DC-based immunotherapy. Studies have shown that polysaccharide can promote DCs function. This study is aimed at exploring the mechanism of LBP affecting DCs function from the perspective of metabolomics. MTT method was used to detect the activity of DC2.4 cells. ELISA kit method was used to detect the contents of IL-6, IL-12, and TNF- in the supernatant of cells. Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to detect general changes in DC2.4 cell metabolism. And then multidistance covariates and bioinformatics, partial least squares-discriminant analysis (PLS-DA) were used to analyze differential metabolites. Finally, metabolic pathway analysis was performed by MetaboAnalyst v5.0. The results showed that LBP had no significant inhibitory effect on the activity of DC2.4 cells at the experimental dose of 50-200 g/ml. LBP (100 g/ml) could significantly stimulate DC2.4 cells to secrete IL-6, TNF-, and IL-12. Moreover, 20 differential metabolites could be identified, including betaine, hypoxanthine, L-carnitine, 5'-methylthioadenosine, orotic acid, sphingomyelin, and L-glutamine. These metabolites were involved 28 metabolic pathways and the top 5 metabolic pathways were aspartate metabolism, pyrimidine metabolism, phenylacetate metabolism, methionine metabolism, and fatty acid metabolism. These results suggest that the effect of LBP on DCs function is related to the regulation of cell metabolism.
靶向树突状细胞 (DC) 代谢相关途径和原位激活 DC 已成为基于 DC 的免疫治疗的新趋势。研究表明，多糖可促进 DC 功能。本研究旨在从代谢组学角度探讨 LPS 影响 DC 功能的机制。MTT 法检测 DC2.4 细胞活性。ELISA 试剂盒法检测细胞上清液中 IL-6、IL-12 和 TNF-的含量。超高效液相色谱-四极杆飞行时间质谱 (UPLC-Q-TOF/MS) 检测 DC2.4 细胞代谢的一般变化。然后采用多距离协变量和生物信息学、偏最小二乘判别分析（PLS-DA）分析差异代谢物。最后，通过 MetaboAnalyst v5.0 进行代谢途径分析。结果表明，在 50-200μg/ml 的实验剂量下，LPS 对 DC2.4 细胞的活性没有明显的抑制作用。LPS（100μg/ml）可显著刺激 DC2.4 细胞分泌 IL-6、TNF-和 IL-12。此外，可鉴定出 20 种差异代谢物，包括甜菜碱、次黄嘌呤、L-肉碱、5'-甲基硫代腺苷、乳清酸、神经鞘磷脂和 L-谷氨酰胺。这些代谢物涉及 28 条代谢途径，前 5 条代谢途径为天冬氨酸代谢、嘧啶代谢、苯乙酸代谢、蛋氨酸代谢和脂肪酸代谢。这些结果表明，LPS 对 DC 功能的影响与细胞代谢的调节有关。
