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  • MiR-1246 Promotes LPS-Induced Inflammatory Injury in Chondrogenic Cells ATDC5 by Targeting HNF4γ.

MiR-1246 Promotes LPS-Induced Inflammatory Injury in Chondrogenic Cells ATDC5 by Targeting HNF4γ.

Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology (2017-10-23)
Da-Peng Wu, Jun-Lei Zhang, Jing-Yu Wang, Ming-Xing Cui, Jin-Ling Jia, Xiang-Hua Liu, Qiu-Dong Liang
ABSTRACT

Osteoarthritis (OA) is a common inflammatory joint disease. miRNAs are associated with OA and functionally implicated in the pathogenesis of the disease. In the present study, we investigated the role of miR-1246 in the lipopolysaccharide (LPS)-induced inflammatory injury of ATDC5 cells. ATDC5 cells were cultured and treated with LPS in a series of concentration (0, 1, 5, and 10 µg/ml) for 5 h. The cells were transfected with miR-1246-mimic, inhibitor, si-HNF4γ or negative control, then were assessed for cell viability using CCK8 assay, apoptosis by flow-cytometry and expressions of miR-1246 and pro-inflammatory cytokines by qRT-PCR and western blot analysis. Cell viability was significantly reduced and cell apoptosis was added in ATDC5 cells injured with LPS at the dosage of 5 and 10 µg/ml. Relative mRNA expressions of pro-inflammatory cytokines (IL-1β, IL-6, IL-8 and TNF-α) were significantly increased. miR-1246 was up-regulated in ATDC5 cells treated with LPS. Moreover, miR-1246 overexpression aggravated LPS-induced decrease in cell viability, increase in apoptosis and overproduction of pro-inflammatory factors. mRNA and protein expressions of HNF4γ were significantly suppressed in cells transfected with miR-124-mimic. Further, miR-1246 knockdown alleviated LPS-induced inflammatory injury by up-regulating the expression of HNF4γ and activation of PI3K/AKT and JAK/STAT pathways. Suppression of miR-1246 alleviated LPS-induced inflammatory injury in chondrogenic ADTC5 cells by up-regulation of HNF4γ and activation of PI3K/AKT and JAK/STAT pathways. The findings of this study will provide a novel viewpoint regarding miR-1246 target for clinical.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-phospho-JAK1 (pTyr1022) antibody produced in rabbit, affinity isolated antibody
Sigma-Aldrich
Glyceraldehyde-3-phosphate Dehydrogenase from rabbit muscle, lyophilized powder, ≥75 units/mg protein