论文部分内容阅读
目的探讨血小板源性生长因子(PDGF)是否通过对细胞外信号调节激酶1/2(ERK1/2)途径的调节进而促进矽肺纤维化的形成与发展。方法采用一次性支气管内灌注二氧化硅(SiO2)粉尘制作矽肺大鼠模型,将其分为对照组(4w组和8w组)、矽肺模型组(4w组和8w组)。采用贴块法进行肺成纤维细胞的原代和传代培养。HE染色观察肺组织形态学变化。羟脯胺酸法检测肺组织内总胶原含量。免疫组化染色检测磷酸化ERK1/2蛋白在肺组织内的分布与表达。Western blot法检测肺组织PDGF及其受体蛋白和ERK1/2及磷酸化ERK1/2蛋白的表达以及肺成纤维细胞Ⅰ型、Ⅲ型胶原蛋白和ERK1/2及磷酸化ERK1/2蛋白的表达。结果与对照组比较,矽肺大鼠肺内PDGF及其受体、磷酸化ERK1/2蛋白表达以及总胶原蛋白含量均增强。在培养的肺成纤维细胞,PDGF能够刺激肺成纤维细胞Ⅰ型、Ⅲ型胶原蛋白表达增加,同时上调磷酸化ERK1/2蛋白的表达,而细胞外信号调节激酶通路特异性抑制剂(PD98059)能够抑制PDGF刺激的肺成纤维细胞增殖与Ⅰ型、Ⅲ型胶原蛋白表达和磷酸化ERK1/2蛋白的表达。结论 PDGF可能通过介导的ERK1/2信号转导通路的激活,进而促进了矽肺纤维化的形成与发展。
Objective To investigate whether platelet derived growth factor (PDGF) promotes the formation and development of silicotic fibrosis by regulating the extracellular signal-regulated kinase 1/2 (ERK1 / 2) pathway. Methods Silicosis rats were made by intraperitoneal infusion of silica dust. The rats were divided into control group (4w group and 8w group) and silicosis model group (4w group and 8w group). The patch method for primary and subculture of lung fibroblasts. The morphological changes of lung tissue were observed by HE staining. Hydroxyproline method was used to detect the total collagen content in lung tissue. The distribution and expression of phosphorylated ERK1 / 2 protein in lung tissue were detected by immunohistochemistry. The expression of PDGF and its receptor protein, ERK1 / 2 and phosphorylated ERK1 / 2 protein and the expression of type Ⅰ and type Ⅲ collagen and ERK1 / 2 and phosphorylated ERK1 / 2 protein in lung fibroblasts were detected by Western blot . Results Compared with the control group, the expression of PDGF and its receptor, phosphorylated ERK1 / 2 protein and total collagen in the lung of silicosis rats increased. In cultured lung fibroblasts, PDGF can stimulate the expression of type I and type III collagen in lung fibroblasts and up-regulate the expression of phosphorylated ERK1 / 2 protein, while the extracellular signal-regulated kinase pathway-specific inhibitor (PD98059) Can inhibit PDGF-stimulated lung fibroblast proliferation and type I, type III collagen protein expression and phosphorylated ERK1 / 2 protein expression. Conclusion PDGF may promote the formation and development of silicotic fibrosis through the activation of ERK1 / 2 signal transduction pathway.