Objective To observe the effect of microRNA (miR)-142-3p on lipopolysaccharide (LPS)-induced inflammatory response in human nasal mucosal epithelial cells (HNEPC) and its targeting relationship with TNF-α.
Methods HNEPC were selected, and an inflammatory response model was established by LPS induction. The miR-142-3p up-regulated, miR-142-3p down-regulated and empty plasmid were transfected into cells, which were divided into control group, LPS group, miR-142-3p up-regulated group, miR-142-3p down-regulated group and empty plasmid group; TNF-α down-regulated and down-regulated control plasmids and miR-142-3p up-regulated plasmids were co-transfected into cells, respectively, and were labeled as miR-142-3p up-regulated +TNF-α down-regulated group, empty plasmid+TNF-α down-regulated group, miR-142-3p up-regulated group and empty plasmid group. Methyl Thiazolyl Tetrazolium (MTT) was used to detect cell proliferation; the real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect the mRNA levels of miR-142-3p, interleukin (IL)-6, IL-10, interferon-γ (IFN-γ) and tumor necrosis factor-alpha (TNF-α); the contents of IL-6, IL-10, IFN-γ and TNF-α in the supernatant were detected by enzyme-linked immunosorbent assay; the targeting relationship between miR-142-3p and TNF-α was analyzed by TargetScan website and luciferase report experiment.
Results The cell proliferation capacity of the LPS group was significantly higher than that of the control group, was significantly higher in the miR-142-3p up-regulated group than that of the empty plasmid group, and was significantly lower in the miR-142-3p down-regulated group than that of the empty plasmid group (P < 0.05). The expression IL-6, IL-10, IFN-γ and TNF-α proteins levels and their mRNA levels in the LPS group were significantly higher than those in the control group, the expression levels of cell-related inflammatory factors in miR-142-3p up-regulated group were significantly higher than those in the empty plasmid group, and the expression level of cell-related inflammatory factors in the miR-142-3p down-regulated group was significantly lower than that in the empty plasmid group (P < 0.05). The mRNA expression levels of IL-6, IL-10, IFN-γ and TNF-α and the protein expression levels of cell supernatant in the TNF-α down-regulated group were significantly lower than those in the down-regulated control group (P < 0.05). MiR-142-3p had a good targeting relationship with TNF-α.
Conclusion The miR-142-3p is highly expressed in nasal mucosa tissues of patients with chronic rhinosinusitis with nasal polyps. Overexpression of miR-142-3p can promote LPS-induced HNEPC inflammatory response, which may be related to up-regulation of gene expression.
DownLoad: