Biochemical and biophysical research communications · 2025
The paradoxical effects of 7-Ketocholesterol and TO901317 on LPS-induced inflammation in human retinal pigment epithelium cells.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Yu Peng, Qiu Yiguo, Mai Yuxin, Shijie Zheng, Yang Lin, Lei Bo
Аннотация
PURPOSE: Liver X receptors (LXRs) are key regulators of cholesterol homeostasis and important modulators of inflammation. Synthetic LXR agonists exert anti-inflammatory effects, whereas endogenous LXR agonist oxysterol has pro-inflammatory effects. This study investigated the paradoxical effects of LXR agonists on inflammation and explored the underlying mechanisms using transcriptomic and proteomic analyses.
METHODS: ARPE-19 cells were incubated with various concentrations of the LXR agonist 7-Ketocholesterol (7-KC) for 36 h. Cell viability and apoptosis were measured by CCK-8 and Annexin-V FITC assays, respectively. Cell proliferation was assessed using EdU assay. RPE cells were pretreated with 7-KC and TO90 for 12 h, followed by stimulation with 5 μg/mL LPS for 24 h. The mRNA levels of IL6, IL-8, LXRα/β, ABCA1, and ABCG1 were measured. Transcriptomic and proteomic profiles were characterized using Illumina HiSeq 2500 and Q Exactive mass spectrometry.
RESULTS: Incubation with high concentrations of 7-KC markedly decreased cell viability and increased apoptosis. Cell proliferation was not observed after treatment with various concentrations of 7-KC for 36 h 7-KC enhanced the mRNA expression of IL-6, IL-8, LXRα, LXRβ, ABCA1, and ABCG1 in a dose-dependent manner. Therefore, 7.5 μM 7-KC was identified as the optimum concentration. Both 7-KC and TO90 activated the LXR target genes ABCA1 and ABCG1. However, 7-KC increased, while TO90 suppressed, IL-6 and IL-8 mRNA levels. Transcriptomic analysis indicated that 7-KC activated toll-like receptor and MAPK signal pathways, whereas TO90 suppressed these pathways. Combined analysis of transcriptomic and proteomic data revealed that 7-KC regulated lipid metabolism and enhanced expression of inflammation-related signaling pathways, including toll-like receptors, MAPK, mTOR, HIF-1, and VEGF pathways. In contrast, TO90 did not directly affect these pathways.
CONCLUSIONS: Both 7-KC and TO90 activated their respective receptor target genes. However, 7-KC amplified LPS-induced inflammation, whereas TO90 inhibited the inflammatory response. These paradoxical effects may result from differential regulation by 7-KC and TO90 on toll-like receptor, HIF-1, VEGF, mTOR, and MAPK signaling pathways, and lipid metabolism-related genes.
METHODS: ARPE-19 cells were incubated with various concentrations of the LXR agonist 7-Ketocholesterol (7-KC) for 36 h. Cell viability and apoptosis were measured by CCK-8 and Annexin-V FITC assays, respectively. Cell proliferation was assessed using EdU assay. RPE cells were pretreated with 7-KC and TO90 for 12 h, followed by stimulation with 5 μg/mL LPS for 24 h. The mRNA levels of IL6, IL-8, LXRα/β, ABCA1, and ABCG1 were measured. Transcriptomic and proteomic profiles were characterized using Illumina HiSeq 2500 and Q Exactive mass spectrometry.
RESULTS: Incubation with high concentrations of 7-KC markedly decreased cell viability and increased apoptosis. Cell proliferation was not observed after treatment with various concentrations of 7-KC for 36 h 7-KC enhanced the mRNA expression of IL-6, IL-8, LXRα, LXRβ, ABCA1, and ABCG1 in a dose-dependent manner. Therefore, 7.5 μM 7-KC was identified as the optimum concentration. Both 7-KC and TO90 activated the LXR target genes ABCA1 and ABCG1. However, 7-KC increased, while TO90 suppressed, IL-6 and IL-8 mRNA levels. Transcriptomic analysis indicated that 7-KC activated toll-like receptor and MAPK signal pathways, whereas TO90 suppressed these pathways. Combined analysis of transcriptomic and proteomic data revealed that 7-KC regulated lipid metabolism and enhanced expression of inflammation-related signaling pathways, including toll-like receptors, MAPK, mTOR, HIF-1, and VEGF pathways. In contrast, TO90 did not directly affect these pathways.
CONCLUSIONS: Both 7-KC and TO90 activated their respective receptor target genes. However, 7-KC amplified LPS-induced inflammation, whereas TO90 inhibited the inflammatory response. These paradoxical effects may result from differential regulation by 7-KC and TO90 on toll-like receptor, HIF-1, VEGF, mTOR, and MAPK signaling pathways, and lipid metabolism-related genes.