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目的研究硬皮病成纤维细胞与纤连蛋白粘附 ,以及粘附诱导的酪氨酸磷酸化蛋白在成纤维细胞表达原胶原mRNA中的作用。方法将硬皮病成纤维细胞置于纤连蛋白包被的培养体系中粘附培养 ,运用RT PCR、免疫印迹法分别检测原胶原mRNA表达的变化及粘附诱导的酪氨酸磷酸化蛋白 ;并观察阻断酪氨酸磷酸化后 ,原胶原mRNA和酪氨酸磷酸化蛋白的变化。结果硬皮病成纤维细胞与纤连蛋白粘附 ,可诱导相对分子质量为98000、65000酪氨酸磷酸化蛋白生成 ,且原胶原 proα1(Ⅰ)mRNA的表达显著增加 ;经酪氨酸激酶抑制剂作用后 ,伴随着相对分子质量为98000、65000酪氨酸磷酸化蛋白的减少 ,原胶原proα1(Ⅰ)mRNA显著降低。结论成纤维细胞与纤连蛋白粘附 ,在硬皮病原胶原表达增加中具有重要作用 ,由粘附诱导的酪氨酸磷酸化是其中的一个重要环节。
Objective To study the adhesion of scleroderma fibroblasts to fibronectin and the role of adhesion-induced tyrosine phosphorylation protein in procollagen mRNA expression in fibroblasts. Methods Scleroderma fibroblasts were cultured in fibronectin - coated culture system. RT PCR and Western blotting were used to detect the changes of procollagen mRNA expression and adhesion - induced tyrosine phosphorylation protein respectively. The changes of procollagen mRNA and tyrosine phosphorylation protein after blocking tyrosine phosphorylation were observed. Results The adhesion of scleroderma fibroblasts to fibronectin induced the formation of tyrosine phosphorylated protein of 98000,65000 and the expression of procollagen α 1 (Ⅰ) mRNA was significantly increased. After tyrosine kinase inhibition After treatment, the pro collagen mRNA of proα1 (Ⅰ) decreased significantly with the decrease of tyrosine phosphorylated protein of 98000 and 65000. Conclusion The adhesion of fibroblasts to fibronectin plays an important role in the increase of the expression of collagen in the dermis, and the tyrosine phosphorylation induced by adhesion is one of the important links.