Xiuchun Wang1, Yali Zhang2. 1. Department of Burn and Plastic Surgery, Jining No.1 People's Hospital, Jining, 272011, Shandong, China. 2. Department of Burn and Plastic Surgery, Jining No.1 People's Hospital, Jining, 272011, Shandong, China. Electronic address: zhangyali0059@sina.com.
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.
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.
Authors: Si Wen; Jiechen Zhang; Bin Yang; Peter M Elias; Mao-Qiang Man Journal: Evid Based Complement Alternat Med Date: 2020-04-14 Impact factor: 2.629