【摘要】：目的:探討不可分型流感嗜血桿菌(nontypeable Haemophilus influenzae,NTHi)誘導(dǎo)氣道上皮細(xì)胞表達(dá)黏蛋白MUC5AC的影響,并探討其可能的分子機(jī)制。方法:體外培養(yǎng)NCI-H292細(xì)胞,用NTHi感染后,采用ELISA檢測(cè)MUC5AC和基質(zhì)金屬蛋白酶9(MMP-9)的水平;明膠酶譜實(shí)驗(yàn)分析MMP-9的酶活性;同時(shí)分別采用表皮生長因子受體(EGFR)、磷脂酰肌醇3-激酶(PI3K)、NADPH氧化酶、活性氧簇(ROS)、和MMP-9特異性抑制劑AG1478、LY294002、DPI、NAC和GM6001預(yù)處理NCI-H292細(xì)胞,檢測(cè)MUC5AC以及MMP-9的水平。結(jié)果:NTHi能以時(shí)間依賴性方式誘導(dǎo)NCI-H292細(xì)胞產(chǎn)生MMP-9,并上調(diào)其酶活性,同時(shí)增加Rac1的活性并誘導(dǎo)ROS生成;采用AG1478和LY294002處理后,Rac1活性顯著降低;采用DPI或Rac1抑制劑NSC23766處理后,ROS含量明顯減少;當(dāng)NCI-H292細(xì)胞用NAC或NSC23766預(yù)處理后,可顯著下調(diào)MMP-9的表達(dá)與活性。此外,采用GM6001處理后,MUC5AC的分泌明顯降低。結(jié)論:NTHi經(jīng)EGFR/PI3K/Rac1/NADPH氧化酶/ROS/MMP-9通路誘導(dǎo)NCIH292細(xì)胞產(chǎn)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.
【作者單位】： 湘南學(xué)院基礎(chǔ)醫(yī)學(xué)部;郴州市第一人民醫(yī)院兒童醫(yī)院;清遠(yuǎn)市獅子湖醫(yī)院;
【基金】：國家自然科學(xué)基金資助項(xiàng)目(No.31300156) 湖南省教育廳計(jì)劃項(xiàng)目(No.09C912) 湘南學(xué)院“十二五”重點(diǎn)學(xué)科(No.xnu125kd019)
【分類號(hào)】：R563.9

【共引文獻(xiàn)】

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