United States Department of Agriculture, Agricultural Research Service, Southern Regional Research Center, New Orleans, LA 70124, USA.
Biomolecules. 2023 Mar 30;13(4):624. doi: 10.3390/biom13040624.
Gossypol is a complex plant polyphenol reported to be cytotoxic and anti-inflammatory, but little is known about its effect on gene expression in macrophages. The objective of this study was to explore gossypol's toxicity and its effect on gene expression involved in the inflammatory response, glucose transport and insulin signaling pathways in mouse macrophages. Mouse RAW264.7 macrophages were treated with multiple concentrations of gossypol for 2-24 h. Gossypol toxicity was estimated by MTT assay and soluble protein content. qPCR analyzed the expression of anti-inflammatory tristetraprolin family (TTP/ZFP36), proinflammatory cytokine, glucose transporter (GLUT) and insulin signaling genes. Cell viability was greatly reduced by gossypol, accompanied with a dramatic reduction in soluble protein content in the cells. Gossypol treatment resulted in an increase in TTP mRNA level by 6-20-fold and increased ZFP36L1, ZFP36L2 and ZFP36L3 mRNA levels by 26-69-fold. Gossypol increased proinflammatory cytokine TNF, COX2, GM-CSF, INFγ and IL12b mRNA levels up to 39-458-fold. Gossypol treatment upregulated mRNA levels of GLUT1, GLUT3 and GLUT4 genes as well as INSR, AKT1, PIK3R1 and LEPR, but not APP genes. This study demonstrated that gossypol induced macrophage death and reduced soluble protein content, which was accompanied with the massive stimulation of anti-inflammatory TTP family and proinflammatory cytokine gene expression, as well as the elevation of gene expression involved in glucose transport and the insulin signaling pathway in mouse macrophages.
棉酚是一种复杂的植物多酚,据报道具有细胞毒性和抗炎作用,但人们对其在巨噬细胞中基因表达的影响知之甚少。本研究旨在探讨棉酚对小鼠巨噬细胞炎症反应、葡萄糖转运和胰岛素信号通路相关基因表达的毒性作用及其影响。用不同浓度的棉酚处理 RAW264.7 巨噬细胞 2-24 小时。MTT 法和可溶性蛋白含量测定棉酚毒性。qPCR 分析抗炎三丝氨酸蛋白家族(TTP/ZFP36)、促炎细胞因子、葡萄糖转运体(GLUT)和胰岛素信号基因的表达。棉酚使细胞活力显著降低,同时细胞内可溶性蛋白含量明显减少。棉酚处理使 TTP mRNA 水平增加 6-20 倍,ZFP36L1、ZFP36L2 和 ZFP36L3 mRNA 水平增加 26-69 倍。棉酚增加促炎细胞因子 TNF、COX2、GM-CSF、INFγ和 IL12b mRNA 水平达 39-458 倍。棉酚处理上调 GLUT1、GLUT3 和 GLUT4 基因以及 INSR、AKT1、PIK3R1 和 LEPR 的 mRNA 水平,但不影响 APP 基因。本研究表明,棉酚诱导巨噬细胞死亡,降低可溶性蛋白含量,同时大量刺激抗炎 TTP 家族和促炎细胞因子基因表达,以及上调小鼠巨噬细胞葡萄糖转运和胰岛素信号通路相关基因的表达。