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目的 探讨蛋白激酶C(PKC)信号通路与温石棉致人胚肺成纤维细胞 (HEPF)的细胞周期及细胞凋亡改变的联系。方法 利用流式细胞技术测定PKC的抑制剂或激活剂对温石棉处理兔肺泡巨噬细胞 (AM)的培养上清液致HEPF的细胞周期和凋亡改变的影响。同时设立空白对照、正常对照 (不加粉尘的AM)、阴性对照 (标准TiO2 )和阳性对照 (标准SiO2 )。结果 温石棉处理AM的上清液刺激HEPF增殖时 ,HEPF的G2 /M期细胞百分比为 7.2 % ,高于各对照组 ,而细胞凋亡百分率为 3.5 % ,低于各对照组 ,差异均有显著性 (P <0 .0 1) ;PKC抑制剂C2 5H2 4 N4 O2 使温石棉所致HEPF增殖时的S期细胞百分比由 37.8%减少到2 7.3% ,凋亡百分率由 3.5 %增加到 2 2 .7% ,差异均有显著性 (P <0 .0 1) ,而PKC激活剂PMA能使此S期细胞百分比增加。结论 温石棉所致HEPF增殖时的细胞周期变化与其上游PKC信号通路有关 ,PKC抑制剂和激活剂主要是作用于S期DNA的合成和诱导细胞凋亡
Objective To investigate the relationship between the protein kinase C (PKC) signaling pathway and the cell cycle and apoptosis of human embryonic lung fibroblasts (HEPF) induced by chrysotile. Methods Flow cytometry was used to determine the effects of inhibitors or activators of PKC on the cell cycle and apoptosis of HEPF induced by chrysotile in cultured alveolar macrophages (AM). At the same time, blank control, normal control (dust-free AM), negative control (standard TiO2) and positive control (standard SiO2) were set up. Results The percentage of G2 / M phase cells in HEPF stimulated by chrysotile in AM supernatant was 7.2%, which was higher than that in control cells, while the percentage of apoptosis was 3.5%, which was lower than that of control group (P <0.01). The PKC inhibitor C2H2 4 N4 O2 decreased the percentages of S phase cells from 37.8% to 23.3% in HEPF induced by chrysotile and the percentage of apoptosis increased from 3.5% to 2 2 .7%, the difference was significant (P <0. 01), while PKC activator PMA increased the percentage of S-phase cells. Conclusion The cell cycle of HEPF induced by chrysotile is related to its upstream PKC signaling pathway. PKC inhibitors and activators mainly act on S phase DNA synthesis and induce apoptosis