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为探讨蛋白激酶在青石棉诱导的肺成纤维细胞增殖中的作用 ,本研究采用体外细胞培养技术 ,用兔肺泡巨噬细胞 (AM)和人胚肺成纤维细胞 (HEPF)组成体外模型 ,用 MTT法测定 HEPF的增殖活力 ,检测了蛋白激酶 (PKA、PKC和 TPK)的抑制剂和激活剂对青石棉诱导的 HEPF增殖的影响 ,以二氧化钛为阴性对照 ,标准石英为阳性对照。结果发现 ,PKA、PKC和 TPK的抑制剂和激活剂均能使青石棉诱导的 HEPF增殖受到抑制和激活 ,并随剂量的变化而变化 ,呈显著的剂量 -效应关系 (P<0 .0 1) ,且青石棉组被抑制和激活的程度强于各对照组。从 3种蛋白激酶的作用强度分析 ,发现 TPK信号通路在青石棉诱导的 HEPF增殖中的作用最强 ,其次是 PKC信号通路 ,PKA信号通路的作用最弱。提示 PKA、PKC和 TPK信号通路均参与了青石棉处理 AM上清液致 HEPF增殖过程 ,TPK信号通路可能起着主导作用 ,这为寻找石棉致纤维化因子的研究方向提供了参考。
In order to investigate the role of protein kinases in the crocidolite-induced proliferation of lung fibroblasts, in vitro cell culture techniques were used to establish in vitro models of rabbit alveolar macrophages (AM) and human embryonic lung fibroblasts (HEPF) MTT method was used to measure the proliferation activity of HEPF. The effects of inhibitors and activators of protein kinases (PKA, PKC and TPK) on crocidolite-induced HEPF proliferation were examined. Titanium dioxide was used as a negative control and standard quartz was used as a positive control. The results showed that inhibitors and activators of PKA, PKC and TPK both inhibited and activated crocidolite-induced proliferation of HEPF, with dose-dependent changes (P <0.01) ), And crocidolite group was inhibited and activated to a greater extent than the control group. From the intensity analysis of the three kinds of protein kinases, it was found that TPK signaling pathway played the strongest role in crocidolite-induced HEPF proliferation, followed by PKC signaling pathway, PKA signaling pathway was the weakest. It is suggested that the PKA, PKC and TPK signal pathways are involved in the HEPF proliferation process induced by crocidolite AM supernatant, and TPK signaling pathway may play a leading role, which provides a reference for the research direction of asbestos-induced fibrosis factor.