Resveratrol alleviates LPS-induced injury in human keratinocyte cell line HaCaT by up-regulation of miR-17
2018
Abstract Background Resveratrol (RSV), an edible polyphenolic phytoalexin, plays an important role in ameliorating inflammation, including skin inflammation after burn injury. However, the specific molecular mechanism underlying its anti-inflammation effect is still unclear. Herein, the effect and the mechanism underlying the protection of HaCaT cells by RSV against inflammation were examined. Methods Lipopolysaccharide (LPS)-induced inflammation and the cytoprotection of RSV were evaluated by detecting viability, apoptosis, expressions of apoptosis-associated proteins and the productions of pro-inflammatory factors by CCK-8 assay, flow cytometer, Western blot, and qRT-PCR. miR-17 expression in RSV-treated HaCaT cells was determined by qRT-PCR. The role of miR-17 in protective effect of RSV was investigated after altering its expression using transfection assay. The main ingredients in PTEN/PI3K/AKT and mTOR pathways were quantified by Western blot. Results LPS-induced HaCaT cell injury was inhibited by RSV administration. RSV promoted viability, inhibited apoptotic cell rate, increased Bcl-2 expression, decreased Bax, cleaved-Caspase-3, and cleaved-Caspase-9 expressions. RSV also inhibited inflammation injury of HaCaT cells by reducing productions of IL-6, IL-8, and TNF-α. miR-17 was up-regulated in LPS and RSV-co-treated cells. The protective effect of RSV might contribute to overexpression of miR-17. In the mechanism study, RSV-miR-17 axis was found to activate PTEN/PI3K/AKT and mTOR pathways in LPS-treated cells. Conclusion RSV alleviated LPS-induced injury in human keratinocyte cell line HaCaT through activations of PTEN/PI3K/AKT and mTOR pathways, which were modulated by miR-17.
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