ERK1／2通路在AcSDKP抑制PDGF诱导的大鼠肺成纤维细胞增殖和胶原合成中的作用  

Role of extracellular signal-regulated kinase1/2 on inhibition of N-aetyl-seryl-asparty-lysyl-proline on proliferation and collagen synthesis of cultured rat pulmonary fibroblasts induced by platelet-derived growth factor

作　　者：武鹍飞 杨方 李丹丹 张丽娟 李倩 王瑞敏 

机构地区：[1]毕北煤炭医学院实验中心,唐山063000

出　　处：《中华劳动卫生职业病杂志》2009年第7期385-389,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：人事部留学人员科技活动资助项目（国人厅发120061164号）;河北省自然科学基金资助项目（C2005000807）;唐山市新药基础研究重点实验室资助项目（04362001B-9）

摘　　要：目的探讨细胞外信号调节激酶（ERK）1／2通路在N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸（AcSDKP）抑制血小板源性生长因子（PDGF）诱导的大鼠肺成纤维细胞增殖和胶原合成中的作用。方法取新生Wistar大鼠20只，获得原代及传代培养的肺成纤维细胞。实验分为：（1）对照组（含体积分数为0．4％的胎牛血清的DMEM组）；（2）PDGF组；（3）PD98059＋PDGF组（25μmol／LPD98059＋10ng／mlPDGF）；（4）AcSDKP＋PDGF组（1×10^8mol／LAcSDKP＋10ng／mlPDGF）。采用噻唑蓝（M1Tr）法检测肺成纤维细胞的代谢活力，免疫细胞化学法、Western blot法检测Ⅰ、Ⅲ型胶原蛋白表达的改变；采用Western blot法检测ERKl／2及磷酸化-ERK1/2蛋白表达的改变。结果与对照组相比，PDGF组大鼠肺成纤维细胞代谢活力增强，Ⅰ、Ⅲ型胶原蛋白表达增强，磷酸化-ERK1/2蛋白表达增高，差异均有统计学意义（P〈0．05）。AcSDKP＋PDGF组细胞代谢活力明显低于PDGF组，免疫细胞化学法检测结果显示，AcSDKP＋PDGF组Ⅰ、Ⅲ型胶原蛋白表达分别为PDGF组的69．3％和67．2％。Westernblot法检测结果显示，AcSDKP＋PDGF组细胞内Ⅰ、Ⅲ型胶原蛋白表达分别为PDGF组的92．4％和78．0％，磷酸化-ERK1／2蛋白表达为PDGF组的83．5％，差异均有统计学意义（P〈0．05）。结论ERK1／2通路在AcSDKP抑制PDGF诱导的大鼠肺成纤维细胞增殖和胶原合成中发挥了重要作用。Objective To investigate the role of extracellular signal-regulated kinase1/2 on the inhibition of N-aetyl-seryl-asparty-lysyl-proline （AcSDKP） on the proliferation and collagen synthesis of cultured rat pulmonary fibroblasts induced by platelet-derived growth factor（PDGF）. Methods Pulmonary fihroblasts were prepared from lungs of neonatal Wistar rats as described previously. Cells were divided into 4 groups： （1） control group （0.4%FBS group） ; （2） PDGF （10 ng/ml）stimulated group; （3）PD98059＋PDGF group （25 μmol/L PD98059＋ 10 ng/ml PDGF）; （ 4 ） AcSDKP＋PDGF group （ 10^-8 mol/L AeSDKP＋ 10 ng/ml PDGF）. All experiments were performed in the fourth passages. Metabolic activity of fibroblasts was observed by MTT, and expressions of type Ⅰ and type Ⅲ collagen were measured by immunocytoehemistry and western blot. Expressions of phospho-ERK1/2 and ERK1/2 were detected by western blot. Results Compared with control group, exposure of pulmonary fibroblasts to 10ng/ml PDGF increased cell metabolic activity, expression of type Ⅰ and type Ⅲ collagen and phosphorylation of ERK1/2. 25p, mol/L PD98059 and AcSDKP both could inhibit the metabolic activity of pulmonary fibroblasts, type Ⅰ and type Ⅲ collagen synthesis and phosphorylation of ERK 1/ 2 induced by PDGF, with significant differences （P〈0.05）. AcSDKP＋PDGF group compared with PDGF stimulated group, metabolic activity of pulmonary fibroblasts decreased to 77.4%. Immunocytochemistry result showed that in AcSDKP＋PDGF group, expressions of type Ⅰ and type Ⅲ collagen decreased to 69.3% and 67.2% compared with those of PDGF stimulated group. Western blot result showed that in AcSDKP ＋PDGF group, expressions of type Ⅰ and type Ⅲ collagen decreased to 92.4% and 78.0%0, phospho-ERK1/2 decreased to 83.5% compared with those of PDGF stimulated group, with significant differences （P〈0.05）. Conclusion ERK1/2 plays an important role in the inhibition of AeSDKP on the proliferation and collagen synth

关 键 词：有丝分裂素激活蛋白激酶类 血小板源生长因子 成纤维细胞 胶原 

分 类 号：R686[医药卫生—骨科学]