蛋白激酶C抑制剂对温石棉诱导肺成纤维细胞增殖的影响

目的 探讨蛋白激酶Ｃ（ＰＣＫ）信号通路在温石棉诱导的肺泡巨噬细胞因子致人胚肺成纤维细胞（ＨＥＰＦ）增殖中的作用。方法 采用体外细胞培养技术，用四甲基偶氮唑蓝（ＭＴＴ）颜色反应法检测温石棉处理兔肺泡巨噬细胞（ＡＭ）的上清液刺激ＨＥＰＦ增殖和ＰＫＣ抑制剂对其产生的影响，同时设空白对照、正常对照（未加粉尘处理的ＡＭ培养上清液）、阴性对照（标准ＴｉＯ２）和阳性对照（标准ＳｉＯ２）。结果 温石棉处理ＡＭ２４ｈ后的培养上清液再处理ＨＥＰＦ４８ｈ，ＨＥＰＦ增殖，且与粉尘浓度存在显著的剂量－效应关系（ｒ＝０．６７１８，Ｐ〈０．０１）；１００ｕｇ／ｍｌ温石棉染尘组ＨＥＰＦ的净增殖率达４０．５６％，是正常对照组的２．６５倍，高于标准ＳｉＯ２组和标准ＴｉＯ２组。１００ｕｇ／ｍｌ粉尘处理ＡＭ的培养上清液的稀释程度与其促使ＨＥＰＦ增殖的

Effect of protein kinase C(PKC) inhibitor on the proliferation of human embryonic pulmonary fibroblasts induced by chrysoffie

Objective To explore the role of protein kinase C(PKC) signal pathway in the proliferation of human embryonic pulmonary fibroblasts (HEPF) caused by chrysotile-induced alveolar macrophage ( AM )-derived factors. Method s MTT color response method to measure the proliferation of HEPF stimulatod by the snpernatants of chrysotile-treated rabbit AMs in vitro was used and the effect of PKC inhibitor on it was observed. Blank control,normal control( AM ), negative ( TiO2 ) and positive control ( SiO2 ) were set up Results The supernatants of chrysetile-treated rabbit AMs significantly stimulated the proliferation of HEPF. The quantity of the proliferation increased with dose-effect relationship( r = 0. 671 8, P < 0.01 ). The net proliferation rate of HEPF in 100μg/ml chrysotile group reached 40.56%, higher than that of SiO2 or TiO2 group, and it was 2.65 times of that of AM group .The higher dilution ratio of the supematants of AM treated by 100 μg/ml dusts was,the less proliferation of HEPF was. Its dose-effect relationship was significant( r = - 0.450 7, P < 0.01 ). Moreover,there was stronger ability to stimulate the proliferation of HEPF at 1: 2 dilution ratio of supernatants in different groups than that of any other dilution ratio. But the degree of proliferation of different experimental dusts was different: chrysotile > SiO2 >TiO2( P < 0.01 ) ,their differences were significant( P < 0.01 ). The proliferation of chrysotile-indnced HEPF was inhibited by PKC inhibitor(C25H24N4O2) with significant dose-effect relationship( P < 0.01 ), and the inhibiting intensity of HEPF in chrysetile group was higher than that in the different control group. Conclusion The supernatants of chrysotile-treated AMs could significantly stimulate the proliferation of HEPF with dose-effect relationship ,while PKC inhibitor could inhibit this effect( P < 0.01 ). There may be some bioactive factors in the supernatant to stimulate the proliferation of HEPF through PKC signal pathway.