β-catenin promotes intracellular bacterial killing via suppression of Pseudomonas aeruginosa-triggered macrophage autophagy.
β-catenin promotes intracellular bacterial killing via suppression of Pseudomonas aeruginosa-triggered macrophage autophagy.
复制标题
β-连环蛋白通过抑制铜绿假单胞菌触发的巨噬细胞自噬来促进细胞内细菌的杀灭。
DOI：
10.1177/0300060517692147
复制
发表时间：
2017-04
期刊：
影响因子：
--
通讯作者：
Wu X
中科院分区：
文献类型：
--
作者：
Fu Q;Chen K;Zhu Q;Wang W;Huang F;Miao L;Wu X
目的探讨β-连环蛋白（β-catenin）在铜绿假单胞菌感染巨噬细胞样细胞RAW 264. 7过程中的作用。通过细胞计数kit-8分析和β-catenin Western印迹检测铜绿假单胞菌感染后RAW 264. 7细胞相对于未感染细胞的细胞活力和catenin beta 1（CTNNB 1）表达。使用流式细胞术和克隆形成有限稀释测定，用β-连环蛋白慢病毒建立CTNNB 1过表达的RAW 264. 7细胞。通过平板计数测量细菌杀伤;通过流式细胞术测量吞噬作用和一氧化氮（NO）;并且使用Griess反应测量活性氧（ROS）。通过微管相关蛋白1轻链3 α-磷脂酰乙醇胺缀合物（LC 3-II）蛋白水平和LC 3斑点的形成，使用Western印迹和免疫荧光染色来确定自噬。铜绿假单胞菌感染后，RAW264.7细胞β-连环蛋白水平以时间和感染复数依赖性方式降低。CTNNB 1过表达与铜绿假单胞菌清除增加相关，但对RAW 264.7细胞吞噬作用、ROS和NO无影响。CTNNB 1过表达降低LC 3-II水平和LC 3斑点的形成，表明自噬抑制。雷帕霉素/饥饿诱导的自噬导致铜绿假单胞菌感染后细菌杀伤减少。β-连环蛋白可能通过抑制铜绿假单胞菌诱导的巨噬细胞自噬来促进细菌杀伤。
To investigate β-catenin-mediated bacterial elimination during Pseudomonas aeruginosa infection of macrophage-like RAW264.7 cells. 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 β-catenin Western blots. RAW264.7 cells with CTNNB1 overexpression were established with β-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. Following P. aeruginosa infection, RAW264.7 cell β-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. β-catenin may promote bacterial killing via suppression of P. aeruginosa-induced macrophage autophagy.
登录
查看更多内容
微生物群对铜绿假单胞菌诱导的角膜炎的抗性。
影响因子：
6.7
作者：
Kugadas A;Christiansen SH;Sankaranarayanan S;Surana NK;Gauguet S;Kunz R;Fichorova R;Vorup-Jensen T;Gadjeva M
通讯作者：
Gadjeva M
Serratia次生代谢产物蛋白质通过产生损害生物分子的活性氧来抑制铜绿假单胞菌生物膜的发育。
影响因子：
5.2
作者：
Kimyon Ö;Das T;Ibugo AI;Kutty SK;Ho KK;Tebben J;Kumar N;Manefield M
通讯作者：
Manefield M
影响因子：
3.7
作者：
Fuerer C;Nusse R
通讯作者：
Nusse R
影响因子：
28.2
作者：
Chen, Kang;Yin, Lin;Huang, Xi
通讯作者：
Huang, Xi
自噬在肺泡巨噬细胞清除铜绿假单胞菌中起着重要作用
影响因子：
4
作者：
Yuan, Kefei;Huang, Canhua;Wu, Min
通讯作者：
Wu, Min