Critical Care Medicine
Role of autophagy in dexmedetomidine-induced reduction of lipopolysaccharide-caused inflammatory responses in macrophages of mice
Xing Mao, Mengying Yan, Hongguang Chen, Jingcheng Feng, Guolin Wang, Keliang Xie, Yonghao Yu
Published 2018-08-20
Cite as Chin J Anesthesiol, 2018, 38(8): 992-995. DOI: 10.3760/cma.j.issn.0254-1416.2018.08.025
Abstract
ObjectiveTo evaluate the role of autophagy in dexmedetomidine-induced reduction of lipopolysaccharide(LPS)-caused inflammatory responses in macrophages of mice.
MethodsMouse macrophage cell line RAW264.7 cultured in vitro were seeded in 6-well or 96-well plates and divided into 4 groups (n=20 each) when cell confluence reached 60% using a random number table method: control group (group Con), LPS group, LPS plus dexmedetomidine group (group LPS+ DEX), and LPS plus dexmedetomidine plus autophagy inhibitor 3-MA group (group LPS+ DEX+ 3-MA). PBS was added and cells were cultured for 12 h in group Con.LPS at the final concentration of 1 000 ng/ml was added and cells were incubated for 12 h in group LPS.LPS at the final concentration of 1 000 ng/ml was added, and then dexmedetomidine at the final concentration of 1 μmol/L was immediately added, and cells were incubated for 12 h in group LPS+ Dex.In group LPS+ Dex+ 3-MA, 3-MA at the final concentration of 2 mmol/L was added and cells were incubated for 1 h, LPS at the final concentration of 1 000 ng/ml was added, and then dexmedetomidine at the final concentration of 1 μmol/L was immediately added, and cells were incubated for 12 h. Cell viability was detected by CCK-8 assay, and the concentrations of nitrous oxide (NO), tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) in the supernatant were determined by enzyme-linked immunosorbent assay, and the expression of microtubule-associated protein 1 light chain 3 Ⅰ (LC3 Ⅰ), LC3Ⅱ, P62 and Bcelin-1 was detected by Western blot.LC3Ⅱ/LC3Ⅰ ratio was calculated.
ResultsCompared with group Con, the cell viability was significantly decreased, the concentrations of TNF-α, IL-1β and NO and LC3Ⅱ/LC3Ⅰ ratio were increased, and the expression of P62 and Beclin1 was up-regulated in group LPS (P<0.05). Compared with group LPS, the cell viability was significantly increased, the concentrations of TNF-α, IL-1β and NO were decreased, LC3Ⅱ/LC3Ⅰ ratio was increased, the expression of P62 was down-regulated, and the expression of Beclin1 was up-regulated in group LPS+ DEX (P<0.05). Compared with group LPS+ Dex, the cell viability was significantly decreased, the concentrations of TNF-α, IL-1β and NO were increased, LC3Ⅱ/LC3Ⅰ ratio was decreased, the expression of P62 was up-regulated, and the expression of Beclin1 was down-regulated in group LPS+ Dex+ 3-MA(P<0.05).
ConclusionEnhanced autophagy is involved in dexmedetomidine-induced reduction of LPS-caused inflammatory responses in macrophages of mice.
Key words:
Dexmedetomidine; Autophagy; Endotoxins; Macrophages; Inflammation
Contributor Information
Xing Mao
Department of Anesthesiology, Tianjin Institute of Anesthesiology General Hospital of Tianjin Medical University, Tianjin 300052, China
Mengying Yan
Department of Anesthesiology, Tianjin Institute of Anesthesiology General Hospital of Tianjin Medical University, Tianjin 300052, China
Hongguang Chen
Department of Anesthesiology, Tianjin Institute of Anesthesiology General Hospital of Tianjin Medical University, Tianjin 300052, China
Jingcheng Feng
Department of Anesthesiology, Tianjin Institute of Anesthesiology General Hospital of Tianjin Medical University, Tianjin 300052, China
Guolin Wang
Department of Anesthesiology, Tianjin Institute of Anesthesiology General Hospital of Tianjin Medical University, Tianjin 300052, China
Keliang Xie
Department of Anesthesiology, Tianjin Institute of Anesthesiology General Hospital of Tianjin Medical University, Tianjin 300052, China
Yonghao Yu
Department of Anesthesiology, Tianjin Institute of Anesthesiology General Hospital of Tianjin Medical University, Tianjin 300052, China
