1Department of Cardiovascular Medicine, Wuhan No. 6 Hospital, Hospital Affiliated to Jianghan University, No. 168, Xianggan Road, Wuhan, 430016 People's Republic of China.
2Department of Pharmacy, Wuhan No. 6 Hospital, Hospital Affiliated to Jianghan University, No. 168, Xianggan Road, Wuhan, 430016 People's Republic of China.
Cell Mol Biol Lett. 2019 Feb 19;24:5. doi: 10.1186/s11658-018-0125-x. eCollection 2019.
Tetrahydroxy stilbene glucoside (TSG) has been reported to exert a cytoprotective effect against various toxicants. However, the function and mechanism of TSG in palmitic acid (PA)-induced inflammation and apoptosis in cardiomyocytes are still unknown. The present study was designed to investigate the post-transcriptional mechanism in TSG-treated cardiomyocytes' inflammation and apoptosis induced by PA.
The mRNA and protein levels were assayed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting, respectively. The targeted genes were predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. Cell proliferation was analyzed by CCK-8 assay. Annexin V-fluorescein isothiocyanate/polyimide (annexin V-FITC/PI) staining was used to evaluate apoptosis using flow cytometry.
TSG restricted the detrimental effects, including the activated inflammatory response and apoptosis, of PA in cardiomyocytes, as well as the up-regulation of miR-129-3p and down-regulation of p-Smad3 expression. In addition, bioinformatics and experimental analysis suggested that Smad3 was a direct target of miR-129-3p, which could inhibit or enhance the expression of p-Smad by transfection with miR-129-3p mimics or inhibitors, respectively. Furthermore, our results demonstrated that overexpression of Smad3 reversed the inhibition of inflammation and apoptosis by overexpression of miR-129-3p in PA-stimulated cardiomyocytes.
TSG targeted to miR-129-3p/Smad3 signaling inhibited PA-induced inflammation and apoptosis in cardiomyocytes.
四羟基二苯乙烯葡萄糖苷（TSG）已被报道对各种毒物具有细胞保护作用。然而，TSG 在软脂酸（PA）诱导的心肌细胞炎症和凋亡中的作用及其机制尚不清楚。本研究旨在探讨 TSG 处理的心肌细胞中 PA 诱导的炎症和凋亡的转录后机制。
采用逆转录定量聚合酶链反应（RT-qPCR）和蛋白质印迹法分别检测 mRNA 和蛋白水平。通过生物信息学算法预测靶向基因，并通过双荧光素酶报告基因检测进行验证。通过 CCK-8 法分析细胞增殖。采用 Annexin V-荧光素异硫氰酸酯/聚酰亚胺（annexin V-FITC/PI）染色，通过流式细胞术评估细胞凋亡。
TSG 抑制了 PA 对心肌细胞的有害作用，包括激活的炎症反应和细胞凋亡，以及 miR-129-3p 的上调和 p-Smad3 表达的下调。此外，生物信息学和实验分析表明，Smad3 是 miR-129-3p 的直接靶点，通过转染 miR-129-3p 模拟物或抑制剂，分别可以抑制或增强 p-Smad 的表达。此外，我们的结果表明，过表达 Smad3 可以逆转 miR-129-3p 过表达对 PA 刺激的心肌细胞中炎症和凋亡的抑制作用。
TSG 通过靶向 miR-129-3p/Smad3 信号通路抑制 PA 诱导的心肌细胞炎症和凋亡。
