Bi Tang1, Ling Xuan1, Mingming Tang1, Hongju Wang1, Jing Zhou1, Jinjun Liu1, Shili Wu1, Miaonan Li1, Xiaojing Wang2, Heng Zhang3. 1. Department of cardiovascular Medicine, The First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, Anhui Province, PR China. 2. Clinical and Basic Provincial Laboratory of Respiratory System Diseases of Anhui Province, The First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, Anhui Province, PR China. 3. Department of cardiovascular Medicine, The First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, Anhui Province, PR China. Electronic address: heng_zhangfh@sina.com.
Abstract
BACKGROUND: miR-93 is recently recognized to perform anti-inflammatory action in the pathological process of cardiomyocytes dysfunction. However, it remains unclear whether miR-93-3p involves in lipopolysaccharide (LPS)-induced inflammation and apoptosis in H9c2 cells. The present study aimed to investigate the functions of miR-93-3p and its target, toll-like receptor 4 (TLR4), in LPS-stimulated cardiomyocytes. MATERIAL AND METHODS: Cell viability was analyzed by CCK-8 assay. AnnexinV-FITC/PI staining and lactate dehydrogenase (LDH) assay were used to evaluate the cell death. The mRNA and protein levels were assayed by RT-qPCR and western blotting, respectively. The targeted gene was predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. RESULTS: LDH stimulation resulted in the suppression of cell viability and the increase in apoptosis rate, inflammatory cytokines and LDH levels, while inhibition of TLR4 with TAK-242 or overexpression of miR-93-3p dramatically blocked LPS-induced inflammation and apoptosis in cardiomyocytes. Intriguingly, bioinformatics analysis and experimental data suggested that TLR4 was a direct target of miR-93-3p, which could inhibit TLR4 expression by transfected with miR-93-3p mimics or elevate the expression of TLR4 by transfected with miR-93-3p inhibitors. Overexpression of TLR4 carried out an opposite effect to miR-93-3p and positively regulated LPS-induced inflammation and apoptosis in cardiomyocytes. CONCLUSION: miR-93-3p showed the protective effects against LPS-induced inflammation and apoptosis in cardiomyocytes by inhibiting TLR4 expression.
BACKGROUND:miR-93 is recently recognized to perform anti-inflammatory action in the pathological process of cardiomyocytes dysfunction. However, it remains unclear whether miR-93-3p involves in lipopolysaccharide (LPS)-induced inflammation and apoptosis in H9c2 cells. The present study aimed to investigate the functions of miR-93-3p and its target, toll-like receptor 4 (TLR4), in LPS-stimulated cardiomyocytes. MATERIAL AND METHODS: Cell viability was analyzed by CCK-8 assay. AnnexinV-FITC/PI staining and lactate dehydrogenase (LDH) assay were used to evaluate the cell death. The mRNA and protein levels were assayed by RT-qPCR and western blotting, respectively. The targeted gene was predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. RESULTS: LDH stimulation resulted in the suppression of cell viability and the increase in apoptosis rate, inflammatory cytokines and LDH levels, while inhibition of TLR4 with TAK-242 or overexpression of miR-93-3p dramatically blocked LPS-induced inflammation and apoptosis in cardiomyocytes. Intriguingly, bioinformatics analysis and experimental data suggested that TLR4 was a direct target of miR-93-3p, which could inhibit TLR4 expression by transfected with miR-93-3p mimics or elevate the expression of TLR4 by transfected with miR-93-3p inhibitors. Overexpression of TLR4 carried out an opposite effect to miR-93-3p and positively regulated LPS-induced inflammation and apoptosis in cardiomyocytes. CONCLUSION:miR-93-3p showed the protective effects against LPS-induced inflammation and apoptosis in cardiomyocytes by inhibiting TLR4 expression.
Authors: Max Hübner; Nicholas Moellhoff; David Effinger; Christian Ludwig Hinske; Simon Hirschberger; Tingting Wu; Martin Bernhard Müller; Gabriele Strauß; Friedrich-Wilhelm Kreth; Simone Kreth Journal: Neurooncol Adv Date: 2020-04-18
Authors: Lu Wang; Jing-Wen Yang; Li-Ting Lin; Jin Huang; Xue-Rui Wang; Xin-Tong Su; Yan Cao; Marc Fisher; Cun-Zhi Liu Journal: Oxid Med Cell Longev Date: 2020-08-11 Impact factor: 6.543