■血管疾病通常是由巨噬细胞和血管平滑肌细胞(VSMC)之间的相互作用引起的。本研究旨在阐明罗格列酮(RSG)的时间疗法是否可以调节巨噬细胞的分泌节律,从而控制VSMC的表型转换并阐明潜在的分子机制,为血管疾病的治疗提供了一种时间治疗方法。
■RAW264.7细胞和A7r5细胞通过50%FBS处理进行同步。通过脂多糖(LPS)暴露诱导M1型巨噬细胞。此外,将靶向PPARγ的siRNA和质粒转染到巨噬细胞中。评估包括细胞活力,迁移,炎症因子水平,脂质代谢物,时钟基因表达,和相对蛋白表达。
■我们透露,与核心时钟基因Bmal1和CLOCK对齐,在ZT2的RSG给药提前了TNF-α释放节律的阶段,而ZT12政府将其向后移动。在ZT2与TNF-α一起孵育显着促进了VSMC的表型转换。在ZT12孵育时,这种作用减弱,这表明VSMC中clock-MAPK途径的参与。此外,在ZT2的RSG给药提前了PPARγ和Bmal1基因的阶段,而ZT12管理将它们向后转移。此外,PPARγ过表达显著诱导巨噬细胞中甘油三酯(TG)积累。外源性TG上调巨噬细胞中Bmal1和CLOCK基因表达并显著增加TNF-α释放。
■涉及RSG的时间疗法诱导巨噬细胞内TG积累,导致昼夜节律基因的改变。这些变化,反过来,调节有节律的TNF-α释放阶段,并在VSMC表型转换中起调节作用。我们的研究为PPARγ激动剂的时间疗法奠定了理论基础。
UNASSIGNED: Vascular diseases are often caused by the interaction between macrophages and vascular smooth muscle cells (VSMCs). This study aims to elucidate whether
chronotherapy with rosiglitazone (RSG) can regulate the secretion rhythm of macrophages, thereby controlling the phenotypic switch of VSMCs and clarifying the potential molecular mechanisms, providing a chronotherapeutic approach for the treatment of vascular diseases.
UNASSIGNED: RAW264.7 cells and A7r5 cells were synchronized via a 50 % FBS treatment. M1-type macrophages were induced through Lipopolysaccharide (LPS) exposure. Additionally, siRNA and plasmids targeting PPARγ were transfected into macrophages. The assessment encompassed cell viability, migration, inflammatory factor levels, lipid metabolites, clock gene expression, and relative protein expression.
UNASSIGNED: We revealed that, in alignment with core clock genes Bmal1 and CLOCK, RSG administration at ZT2 advanced the phase of TNF-α release rhythm, while ZT12 administration shifted it backward. Incubation with TNF-α at ZT2 significantly promoted the phenotype switch of VSMCs. This effect diminished when incubated at ZT12, implicating the involvement of the clock-MAPK pathway in VSMCs. Furthermore, RSG administration at ZT2 advanced the phases of PPARγ and Bmal1 genes, whereas ZT12 administration shifted them backward. Additionally, PPARγ overexpression significantly induced triglyceride (TG) accumulation in macrophages. Exogenous TG upregulated Bmal1 and CLOCK gene expression in macrophages and significantly increased TNF-α release.
UNASSIGNED: Chronotherapy involving RSG induces TG accumulation within macrophages, resulting in alterations in circadian gene rhythms. These changes, in turn, modulate the phase of rhythmic TNF-α release and play a regulatory role in VSMCs phenotype switch. Our study establishes a theoretical foundation for
chronotherapy of PPARγ agonists.