Enhanced Expression of miR-34a Enhances Escherichia coli Lipopolysaccharide-Mediated Endometritis by Targeting LGR4 to Activate the NF-κB Pathway

Figure 3

LGR4 suppresses the secretion of inflammatory cytokines in BENDs. (a) BENDs were treated with LPS (0, 0.5, 1.0, 1.5, and 2.0 μg/ml) for 6 h. The LGR4 mRNA levels were analysed. (b) Cells were stimulated with LPS (1.0 μg/ml) at 0, 3, 6, 12, and 24 h. The expression level of LGR4 was measured by qRT-PCR. (c) The effects of LPS (1.0 μg/ml) on the viability of BENDs. Cell viability was determined with a CCK-8 assay kit. (d, e) Western blot analysis of LGR4 protein in BENDs after treatment with si-LGR4 or negative control (si-NC). β-Actin was used as an internal control. (f) qRT-PCR analysis of LGR4 mRNA expression in BENDs after treatment with si-LGR4 or negative control (si-NC). GAPDH was used as a control. (g) Cells were transfected with si-LGR4 or si-NC for 24 h and then stimulated with LPS (1.0 μg/ml) for 6 h. The production of IL-1β, IL-6, and TNF-α was measured with qRT-PCR. GAPDH was used as a control. The data are presented as the of three independent experiments (). and versus the si-NC group; ^# and ^## versus the si-NC and LPS groups (cells transfected with si-NC after stimulation with LPS) (Student’s t-test).

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