Jian Shang1, Lei Wang2, Lili Tan2, Ren Pan3, Dan Wu3, Yanfei Xia4, Peng Xu5. 1. Department of Anathesiology, People's Hospital of Anji, China. 2. Department of Anathesiology,Gansu Provincial Maternity and Child Care Hospital, China. 3. Department of Anesthesiology, Zhejiang Hospital, China. 4. Department of Anesthesiology, Zhejiang Hospital, China. Electronic address: yanfeixia_yanfei@163.com. 5. Department of Anesthesiology, Zhejiang Hospital, China. Electronic address: pengxu_pex@163.com.
Abstract
BACKGROUND: Acute respiratory distress syndrome (ARDS) is multiple inflammatory injury lung disease. MiR-27a-3p alleviates lung injury, whether miR-27a-3p could affect the lung inflammation is not clear. Therefore, we established the lipopolysaccharides (LPS)-induced alveolar epithelial cell model to simulate ARDS inflammation in vitro to investigate the effect of miR-27a-3p in ARDS. METHODS: After LPS-induced alveolar epithelial cell model was established and FOXO3 was proved to be targeted by miR-27a-3p, the miR-27a-3p mimic, inhibitor, or FOXO3-overexpression plasmids were transfected into the cells. The effects of miR-27a-3p and FOXO3 on cell viability and apoptosis were then evaluated. The levels of apoptosis-/inflammation-related factors, miR-27a-3p, and FOXO3 were further analyzed. Also, the activities of reactive oxygen species (ROS) and nicotinamide adenine dinucleotide phosphate (NAPDH) in cells were examined. RESULTS: MiR-27a-3p was down-regulated in LPS-induced alveolar epithelial cells. The decreased-cell viability of the LPS-induced cells was increased by miR-27a-3p mimic while inhibited by FOXO3. The enhanced-apoptosis, and up-regulated Bax and C caspase-3 were reduced by miR-27a-3p mimic while inhibited by FOXO3; the down-regulated Bcl-2 of the LPS-induced cells was increased by miR-27a-3p mimic while inhibited by FOXO3. The up-regulated IL-6, IL-8, ROS, and NAPDH in the LPS-induced cells were reduced by miR-27a-3p mimic while inhibited by FOXO3. Besides, FOXO3 reversed the effect of miR-27a-3p mimic on the LPS-induced cells. CONCLUSION: MiR-27a-3p targeted FOXO3 to mitigated inflammation and apoptosis of LPS-induced alveolar epithelial cells via suppressing NAPDH/ROS activation.
BACKGROUND:Acute respiratory distress syndrome (ARDS) is multiple inflammatory injury lung disease. MiR-27a-3p alleviates lung injury, whether miR-27a-3p could affect the lung inflammation is not clear. Therefore, we established the lipopolysaccharides (LPS)-induced alveolar epithelial cell model to simulate ARDS inflammation in vitro to investigate the effect of miR-27a-3p in ARDS. METHODS: After LPS-induced alveolar epithelial cell model was established and FOXO3 was proved to be targeted by miR-27a-3p, the miR-27a-3p mimic, inhibitor, or FOXO3-overexpression plasmids were transfected into the cells. The effects of miR-27a-3p and FOXO3 on cell viability and apoptosis were then evaluated. The levels of apoptosis-/inflammation-related factors, miR-27a-3p, and FOXO3 were further analyzed. Also, the activities of reactive oxygen species (ROS) and nicotinamide adenine dinucleotide phosphate (NAPDH) in cells were examined. RESULTS:MiR-27a-3p was down-regulated in LPS-induced alveolar epithelial cells. The decreased-cell viability of the LPS-induced cells was increased by miR-27a-3p mimic while inhibited by FOXO3. The enhanced-apoptosis, and up-regulated Bax and C caspase-3 were reduced by miR-27a-3p mimic while inhibited by FOXO3; the down-regulated Bcl-2 of the LPS-induced cells was increased by miR-27a-3p mimic while inhibited by FOXO3. The up-regulated IL-6, IL-8, ROS, and NAPDH in the LPS-induced cells were reduced by miR-27a-3p mimic while inhibited by FOXO3. Besides, FOXO3 reversed the effect of miR-27a-3p mimic on the LPS-induced cells. CONCLUSION:MiR-27a-3p targeted FOXO3 to mitigated inflammation and apoptosis of LPS-induced alveolar epithelial cells via suppressing NAPDH/ROS activation.