不可分型流感嗜血杆菌诱导NCI-H292细胞产生MUC5AC的分子机制
发布时间：2019-03-03 19:10
【摘要】：目的:探讨不可分型流感嗜血杆菌(nontypeable Haemophilus influenzae,NTHi)诱导气道上皮细胞表达黏蛋白MUC5AC的影响,并探讨其可能的分子机制。方法:体外培养NCI-H292细胞,用NTHi感染后,采用ELISA检测MUC5AC和基质金属蛋白酶9(MMP-9)的水平;明胶酶谱实验分析MMP-9的酶活性;同时分别采用表皮生长因子受体(EGFR)、磷脂酰肌醇3-激酶(PI3K)、NADPH氧化酶、活性氧簇(ROS)、和MMP-9特异性抑制剂AG1478、LY294002、DPI、NAC和GM6001预处理NCI-H292细胞,检测MUC5AC以及MMP-9的水平。结果:NTHi能以时间依赖性方式诱导NCI-H292细胞产生MMP-9,并上调其酶活性,同时增加Rac1的活性并诱导ROS生成;采用AG1478和LY294002处理后,Rac1活性显著降低;采用DPI或Rac1抑制剂NSC23766处理后,ROS含量明显减少;当NCI-H292细胞用NAC或NSC23766预处理后,可显著下调MMP-9的表达与活性。此外,采用GM6001处理后,MUC5AC的分泌明显降低。结论:NTHi经EGFR/PI3K/Rac1/NADPH氧化酶/ROS/MMP-9通路诱导NCIH292细胞产生MUC5AC。
[Abstract]:Aim: to investigate the effect of Haemophilus influenzae (nontypeable Haemophilus influenzae,NTHi (Haemophilus influenzae) on the expression of mucin MUC5AC in airway epithelial cells and its possible molecular mechanism. Methods: NCI-H292 cells were cultured in vitro. After infected with NTHi, the levels of MUC5AC and MMP-9 were detected by ELISA, and the enzyme activity of MMP-9 was analyzed by gelatin zymogram. NCI-H292 cells were pretreated with epidermal growth factor receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), NADPH oxidase, active oxygen cluster (ROS),) and MMP-9 specific inhibitor AG1478,LY294002,DPI,NAC and GM6001, respectively. The levels of MUC5AC and MMP-9 were detected. Results: NTHi could induce MMP-9, production and up-regulate the enzyme activity of NCI-H292 cells in a time-dependent manner, and at the same time increase the activity of Rac1 and induce the production of ROS. After treatment with AG1478 and LY294002, the activity of Rac1 decreased significantly. When NCI-H292 cells were pretreated with NAC or NSC23766, the expression and activity of MMP-9 were significantly down-regulated when treated with DPI or Rac1 inhibitor NSC23766. In addition, after GM6001 treatment, the secretion of MUC5AC decreased significantly. Conclusion: NTHi induces MUC5AC. production in NCIH292 cells via EGFR/PI3K/Rac1/NADPH oxidase / ROS/MMP-9 pathway.
【作者单位】： 湘南学院基础医学部;郴州市第一人民医院儿童医院;清远市狮子湖医院;
【基金】：国家自然科学基金资助项目(No.31300156) 湖南省教育厅计划项目(No.09C912) 湘南学院“十二五”重点学科(No.xnu125kd019)
【分类号】：R563.9
[Abstract]:Aim: to investigate the effect of Haemophilus influenzae (nontypeable Haemophilus influenzae,NTHi (Haemophilus influenzae) on the expression of mucin MUC5AC in airway epithelial cells and its possible molecular mechanism. Methods: NCI-H292 cells were cultured in vitro. After infected with NTHi, the levels of MUC5AC and MMP-9 were detected by ELISA, and the enzyme activity of MMP-9 was analyzed by gelatin zymogram. NCI-H292 cells were pretreated with epidermal growth factor receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), NADPH oxidase, active oxygen cluster (ROS),) and MMP-9 specific inhibitor AG1478,LY294002,DPI,NAC and GM6001, respectively. The levels of MUC5AC and MMP-9 were detected. Results: NTHi could induce MMP-9, production and up-regulate the enzyme activity of NCI-H292 cells in a time-dependent manner, and at the same time increase the activity of Rac1 and induce the production of ROS. After treatment with AG1478 and LY294002, the activity of Rac1 decreased significantly. When NCI-H292 cells were pretreated with NAC or NSC23766, the expression and activity of MMP-9 were significantly down-regulated when treated with DPI or Rac1 inhibitor NSC23766. In addition, after GM6001 treatment, the secretion of MUC5AC decreased significantly. Conclusion: NTHi induces MUC5AC. production in NCIH292 cells via EGFR/PI3K/Rac1/NADPH oxidase / ROS/MMP-9 pathway.
【作者单位】： 湘南学院基础医学部;郴州市第一人民医院儿童医院;清远市狮子湖医院;
【基金】：国家自然科学基金资助项目(No.31300156) 湖南省教育厅计划项目(No.09C912) 湘南学院“十二五”重点学科(No.xnu125kd019)
【分类号】：R563.9
【共引文献】
相关期刊论文 前10条
1 张姗姗;王彦;李德东;曹永兵;姜远英;;NADH和NADPH代谢和功能的研究进展[J];第二军医大学学报;2011年11期
2 钟贞;邓s，
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