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find Keyword "Src" 2 results
  • 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

    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.

    Release date:2016-09-14 11:23 Export PDF Favorites Scan
  • Src ACTIVATION REQUIRED FOR pERK1/2 ACTIVATION IN FOCAL ADHESIONS IN OSTEOBLASTS INDUCED BY PLATELETDERIVED GROWTH FACTOR

    Objective To study the function of platelet-derived growth factor (PDGF) in inducing phosphorylation extracellular signalregulated kinase 1/2 (pERK1/2) localization in osteoblasts. Methods Primary osteoblasts were isolated and cultured from cranial bone of 10 mice atthe age of 3 days, weighting 6-9 g without limitation in male and female.The sixth passage osteoblasts were incubated in 1% serum for 12 hours and divided into 2 groups: treated with DMSO(control group) or with PP2(experimentalgroup) for 30 minutes. Each group was further divided into 2 subgroups according to with or without PDGF (20ng/ml) stimulation for 10 minutes. pERK1/2 localization was analysized by immunofluorescence staining in osteoblasts pretreated with or without Src inhibitor PP2. The sixth passage osteoblasts were divided into 2 groups treated with DMSO (control group) or with PP2 (experimental group) for 30 minutes. Each group was further divided into two subgroupsaccording to with or without PDGF (20 ng/ml) stimulation for 10 mintues. The ability of osteoblast migration was determined by wound healing assay. The sixth passage osteoblasts were divided into 2 groups treated with DMSO (control group) or 10 μmol/L PP2 (experimental group) for 30 mintues. Each group was further divided into 2 subgroups according to with or without PDGF (20 ng/ml) stimulation.The pERK1/2 was determined by Western blot in osteoblastic cytoskeleton inducedby PDGF. Results Immunofluorescence staining showed pERK1/2 localization in osteoblastic nuclears and focal adhesions after PDGF stimulation. PP2 significantly inhibited ERK1/2 localization in focal adhesions, but not in nuclears. The wound healing assay results showed that PP2 significantly inhibited osteoblast migration induced by PDGF. The result of Western blot demonstrated that pERK1/2 in osteoblastic cytoskeleton was significantly inhibitedSrc activation is required for pERK1/2 translocalization to focal adhesions and osteoblasts migration. 

    Release date:2016-09-01 09:20 Export PDF Favorites Scan
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