Aggregatibacter actinomycetemcomitans Invasion Induces Interleukin-1β Production Through Reactive Oxygen Species and Cathepsin B
Aggregatibacter actinomycetemcomitans Invasion Induces Interleukin-1β Production Through Reactive Oxygen Species and Cathepsin B
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DOI：
10.1089/jir.2014.0127
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发表时间：
2015-06-01
影响因子：
2.3
通讯作者：
Nishihara, Tatsuji
中科院分区：
文献类型：
--
作者：
Okinaga, Toshinori;Ariyoshi, Wataru;Nishihara, Tatsuji
白细胞介素-1（IL-1）细胞因子IL-1、IL-1和IL-18在包括牙周炎在内的多种疾病的炎症反应中起着至关重要的作用。在这项研究中，牙周病细菌病原体，放线菌伴聚合杆菌，诱导巨噬细胞的细胞死亡和细胞因子的释放。A.用溴化（3-[4，5-二甲基噻唑-2-基]-2，5-二苯基四唑鎓测定法测定伴放线菌的侵袭。A.用实时荧光定量逆转录聚合酶链反应、蛋白质印迹和酶联免疫吸附试验检测放线菌共生体侵入的巨噬细胞。caspase-1抑制剂处理和caspase-1基因沉默对A.伴放线菌入侵模式识别受体NLRP 3在A.伴放线菌侵入巨噬细胞。然而，NLRP 3敲低对A.伴放线菌侵染RAW 264细胞。此外，A.伴放线菌侵袭RAW 264细胞后，细胞内活性氧（ROS）的产生和组织蛋白酶B的释放均受到抑制。组织蛋白酶B抑制剂CA 074-Me和活性氧抑制剂N-乙酰-L-半胱氨酸可抑制A.伴放线菌综上所述，这些结果表明A.伴随放线菌通过ROS和组织蛋白酶B的产生诱导RAW 264细胞产生IL-1，但不通过NLRP 3/caspase-1途径。
Interleukin-1 (IL-1) cytokines, IL-1, IL-1, and IL-18 play a crucial role in inflammatory responses in a variety of diseases including periodontitis. In this study, the periodontopathic bacterial pathogen, Aggregatibacter actinomycetemcomitans, induced cell death and cytokine release in macrophages. Cell viability was reduced by A. actinomycetemcomitans invasion using (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide assay. The production of IL-1 in A. actinomycetemcomitans-invaded macrophage cells was detected by real-time reverse transcriptase-polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay. Treatment with a caspase-1 inhibitor and silencing of the caspase-1 gene had no effect on IL-1 secretion induced by A. actinomycetemcomitans invasion. Pattern recognition receptor, NLRP3 was upregulated in A. actinomycetemcomitans-invaded macrophages. However, NLRP3 knockdown had no effect on the secretion of IL-1 in A. actinomycetemcomitans-invaded RAW 264 cells. In addition, A. actinomycetemcomitans invasion induced the generation of reactive oxygen species (ROS) and the release of cathepsin B in RAW 264 cells. Interestingly, CA074-Me, a cathepsin B inhibitor, and N-Acetyl-l-cysteine, a ROS inhibitor, prevented the production of IL-1 induced by A. actinomycetemcomitans. Taken together, these results suggest A. actinomycetemcomitans induce IL-1 production in RAW 264 cells through the production of ROS and cathepsin B, but not through the NLRP3/caspase-1 pathway.