期刊
JOURNAL OF INTERNATIONAL MEDICAL RESEARCH
卷 45, 期 2, 页码 556-569出版社
SAGE PUBLICATIONS LTD
DOI: 10.1177/0300060517692147
关键词
Pseudomonas aeruginosa; -catenin; bacterial killing; autophagy
资金
- Guangdong Medical Science Foundation [B2014447]
- National Natural Science Foundation of China [81401645]
- Zhongshan Medical and Health Major Projects Foundation [2016B1001]
Objective To investigate -catenin-mediated bacterial elimination during Pseudomonas aeruginosa infection of macrophage-like RAW264.7 cells. Methods Cell viability and catenin beta 1 (CTNNB1) expression in RAW264.7 cells following P. aeruginosa infection versus uninfected cells, were detected by cell counting kit-8 assay and -catenin Western blots. RAW264.7 cells with CTNNB1 overexpression were established with -catenin lentivirus using flow cytometry and clonogenic limiting dilution assays. Bacterial killing was measured by plate counts; phagocytosis and nitric oxide (NO) were measured by flow cytometry; and reactive oxygen species (ROS) were measured using Griess reaction. Autophagy was determined by microtubule-associated protein 1 light chain 3 alpha-phosphatidylethanolamine conjugate (LC3-II) protein levels and formation of LC3 puncta, using Western blot and immunofluorescence staining. Results Following P. aeruginosa infection, RAW264.7 cell -catenin levels were reduced in a time- and multiplicity of infection-dependent manner. CTNNB1 overexpression was associated with increased P. aeruginosa elimination, but had no effect on RAW264.7 cell phagocytosis, ROS and NO. CTNNB1 overexpression reduced LC3-II levels and formation of LC3 puncta, suggesting autophagy inhibition. Rapamycin/starvation-induced autophagy resulted in reduced bacterial killing following P. aeruginosa infection. Conclusion -catenin may promote bacterial killing via suppression of P. aeruginosa-induced macrophage autophagy.
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