Journal
PATHOLOGY RESEARCH AND PRACTICE
Volume 214, Issue 10, Pages 1686-1693Publisher
ELSEVIER GMBH
DOI: 10.1016/j.prp.2018.08.024
Keywords
Cardiomyocytes; Apoptosis; LPS; miR-93-3p; TLR4
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Funding
- National Natural Science Foundation of China [81170046]
- Key Project of Top-Notch Talent of Discipline (specialty) of the Higher Education Institute of Anhui Province 2016 [gxbjZD2016072]
- Major projects of Natural Science Research in Colleges and Universities of Anhui Province [KJ2018ZD023]
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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.
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