中国呼吸与危重监护杂志

中国呼吸与危重监护杂志

ROS/Src /JNK 信号通路在香烟诱导气道上皮细胞黏液高分泌中的作用

查看全文

目的 探讨在香烟诱导气道上皮细胞黏蛋白MUC5AC 表达过程中Src / JNK 信号通路的作用。方法 香烟抽提物预处理的人气道上皮A549 细胞, 分别以活性氧( ROS) 清除剂DMTU、JNK 特异性抑制剂SP600125 及Src 激酶抑制剂PP2 干预。测定各组细胞中ROS 的含量, 采用RT-PCR、ELISA 法观察细胞黏蛋白( MUC) 5AC转录水平及培养上清液中MUC5AC 蛋白水平的改变,以Western blot 法检测细胞表皮生长因子受体( EGFR) 的蛋白表达。结果 细胞暴露于不同浓度的香烟抽提物中, ROS的量呈浓度递增; ROS清除剂DMTU 显著降低香烟所致的Src 磷酸化; Src 特异性抑制剂PP2 处理组中的JNK 磷酸化水平与对照组比较有明显下降, MUC5AC 的表达水平也明显降低; JNK特异性抑制剂SP600125 组中MUC5AC 的蛋白表达和基因转录水平较对照组均明显降低。结论 ROS-Src-JNK信号通路可能参与A549 上皮细胞中MUC5AC 的表达调控。

Objective To explore the role of ROS/ Src / JNK signaling pathway in cigarette smoke extract( CSE) -induced mucin ( MUC) 5AC production in A549 airway epithelial cells. Methods The A549 airway epithelial cells were cultured in medium with CSE, then treated with ROS scavenger DMTU, c-Jun Nterminal kinase( JNK) specific inhibitor SP600125, and Src kinase inhibitor PP2, respectively. The relative content of reactive oxygen species( ROS) were assayed by special kit. The levels of MUC5AC in culture medium, epidermal growth factor receptor( EGFR) , activated EGFR and MUC5AC mRNA in culture cells were detected with ELISA,Western Blot and RT-PCR, respectively. Results A dose-dependent increasing of ROS production in cells exposed to dilutions of cigarette smoke solution was detected. DMTU inhibited cigarette smoke-induced Src phosphorylation( P lt; 0. 05) . SP600125 reduced the expression of MUC5AC ( P lt; 0. 05) compared with the normal group. The activation of JNK was suppressed by Src specific inhibitor PP2( P lt; 0. 05) . Conclusion ROS/ Src / JNK signal cascade may play a particular role in MUC5AC expression of A549 cells induced by cigarette smoke.

关键词: 黏蛋白; 香烟; 活性氧; Src; JNK

Key words: Mucin; Cigarette smoke; Reactive oxygen species; Src; JNK

引用本文: 田川,李琪,周向东. ROS/Src /JNK 信号通路在香烟诱导气道上皮细胞黏液高分泌中的作用. 中国呼吸与危重监护杂志, 2009, 09(2): 127-130. doi: 复制

登录后 ,请手动点击刷新查看全文内容。 没有账号,
1. by Hydrogen Peroxide in Endothelial Cells Involves Src-dependent Epidermal Growth Factor Receptor Transactivation. J Biol Chem,2001, 276: 16045-16050.
2. Psarras S, Caramori G, Contoli M, et al. Oxidants in asthma and in chronic obstructive pulmonary disease( COPD) . Curr Pharm Des,2005, 11: 2053-2062.
3. Shao MX, Nakanaga T, Nadel JA. Cigarette smoke induces MUC5AC mucin overproduction via tumor necrosis factor-alpha-converting enzyme in human airway epithelial( NCI-H292 ) cells. Am J Physiol Lung Cell Mol Physiol, 2004, 287: L420-L427.
4. Takeyama K, Jung B, Shim JJ, et al. Activation of epidermal growth factor receptors is responsible for mucin synthesis induced by cigarette smoke. Am J Physiol Lung Cell Mol Physiol, 2001, 280:L165-L172.
5. 周向东, 童瑾, 兰箭. 慢性阻塞性肺疾病患者气道黏蛋白分子表型的研究. 中华结核和呼吸杂志, 2002, 25: 437.
6. YoshizumM, Abe JI, Haendeler J, et al. Src and Cas Mediate JNK Activation but Not ERK1 /2 and p38 Kinases by Reactive Oxygen Species. J Biol Chem, 2000, 275: 11706-11712.
7. Nakamura Y, Romberger DJ, Tate L, et al. Cigarette smoke inhibits lung fibroblast proliferation and chemotaxis. Am J Respir Crit Care Med, 1995, 151: 1497-1503.
8. Ma C, Bower KA, Lin H, et al. The role of epidermal growth factor receptor in ethanol-mediated inhibition of activator protein-1 transactivaion. Biochem Pharmacol, 2005, 69 : 1785 -1794 .
9. Pinkus R, Weiner LM, Daniel V. Role of oxidants and antioxidants in the induction of AP-1, NF-κB, and gutathione-s transferase gene expression. J Biol Chem, 1996, 271: 13422-13429.
10. Chen K, Vita JA, Berk BC, et al. c-Jun N-terminal Kinase Activation
11. Meurer S, Pioch S, Gross S. Reactive Oxygen Species Induce Tyrosine Phosphorylation of and Src Kinase Recruitment to NOsensitive Guanylyl Cyclase. J Biol Chen, 2005, 280: 33149-33156.
12. Li JD, Feng W, Gallup M. et al. Activation of NF-κB via a Src-dependent Ras-MAPK-pp90rsk pathway is required for Pseudomonas aeruginosa-induced mucin overproduction in epithelial cells. Proc Natl Acad Sci USA, 1998, 95: 5718-5723.