β-Catenin promotes host resistance against Pseudomonas aeruginosa keratitis
β-Catenin promotes host resistance against Pseudomonas aeruginosa keratitis
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DOI：
10.1016/j.jinf.2013.07.025
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发表时间：
2013-12-01
影响因子：
28.2
通讯作者：
Huang, Xi
中科院分区：
文献类型：
--
作者：
Chen, Kang;Yin, Lin;Huang, Xi
目的：探讨β-连环蛋白在铜绿假单胞菌(PA)角膜炎中的作用。方法：采用免疫印迹和免疫组织化学方法检测C57BL/6(B6)角膜及体外培养的小鼠角膜上皮细胞、巨噬细胞样RAW264.7细胞、骨髓源性中性粒细胞和A6(1)角膜上皮细胞中β-连环蛋白的表达。B6小鼠结膜下注射表达β-连环蛋白活性突变体的慢病毒(β-CAT-慢病毒)和相应的对照(CTL-慢病毒)，然后感染PA。结果：PA感染B6角膜、小鼠巨噬细胞样RAW264.7细胞、小鼠骨髓源性中性粒细胞和小鼠A6(1)角膜上皮细胞后，β-catenin蛋白表达降低。在B6角膜中过表达的β-连环蛋白通过减少促炎细胞因子的表达和细菌负荷，显著降低了PA感染后角膜疾病的严重程度。体外实验数据进一步证明，β-连环蛋白的过度表达抑制了促炎细胞因子的产生，但促进了巨噬细胞和中性粒细胞的细菌清除。结论：β-连环蛋白通过减少角膜炎症和细菌负荷而降低了PA角膜炎的严重程度。(C)2013年英国感染协会。爱思唯尔有限公司出版。保留所有权利。
Objective: To explore the role of beta-catenin in Pseudomonas aeruginosa (PA) keratitis.Methods: Western-blot and immunostaining assay were used to determine the beta-catenin protein expression in C57BL/6 (B6) corneas and in in vitro cultured murine cells including macrophage-like RAW264.7 cells, bone marrow-derived neutrophils and A6(1) corneal epithelial cells. B6 mice were subconjunctivally injected with lentivirus expressing active mutant of beta-catenin (beta-cat-lentivirus) vs appropriate control (Ctl-lentivirus), and then infected with PA. Pro-inflammatory cytokine levels were examined using real-time PCR and ELISA, and bacterial burden was assessed using plate count assays both in vivo and in vitro.Results: beta-Catenin protein expression was decreased in B6 corneas, murine macrophage-like RAW264.7 cells, mouse bone marrow-derived neutrophils and mouse A6(1) corneal epithelial cells after PA infection. Over-expression of beta-catenin in B6 corneas significantly reduced the severity of corneal disease after PA infection, by decreasing pro-inflammatory cytokine expression and bacterial burden. In vitro data further demonstrated that over-expression of beta-catenin suppressed pro-inflammatory cytokine production but enhanced bacterial clearance in macrophages and neutrophils.Conclusions: beta-Catenin reduces the severity of PA keratitis by decreasing corneal inflammation and bacterial burden.(C) 2013 The British Infection Association. Published by Elsevier Ltd. All rights reserved.