可溶型白细胞介素-13受体α2对成纤维细胞胶原蛋白合成的抑制作用

Inhibitory Effect of Soluble IL-13 Receptor Alpha 2 on Fibroblast Collagen Synthesis

目的应用可溶型IL-13受体α2(sIL-13Rα2)抑制成纤维细胞胶原合成,探索生物治疗纤维化的新途径。方法构建pMSCV/sIL-13Rα2载体和表达sIL-13Rα2的细胞模型,分子量(0.2×106)滤膜离心浓缩培养液,Ni2亲和层析,SDS-PAGE分析sIL-13Rα2,Western blot检测sIL-13Rα2,RT-PCR和羟脯氨酸法显示成纤维细胞Ⅰ型胶原蛋白的表达。结果构建的重组体pMSCV/sIL-13Rα2成功转染入HEK293细胞,并表达sIL-13Rα2。50μg.L-1 IL-13促进人瘢痕成纤维细胞系CRL-1762生成Ⅰ型胶原蛋白,50μmol.L-1 sIL-13Rα2能有效抑制IL-13刺激CRL-1762细胞合成Ⅰ型胶原蛋白。结论 sIL-13Rα2靶向IL-13抑制成纤维细胞Ⅰ型胶原蛋白的合成。

Objective To explore a new biological therapy in fibrosis through inhibiting fibroblast collagen synthesis by soluble IL-13 receptor α 2（sIL-13Rα2）.Methods The pMSCV/sIL-13 Rα2 vector was constructed and cell model expressing sIL-13 Rα2 was established.The medium was concentrated by ultrafiltration（0.2×106-molecular-weigh out off） and sIL-13 Rα2 was analyzed by Ni2 affinity chromatography,SDS-PAGE and Western blot.The expression of type Ⅰ collagen protein in fibroblasts was detected by RT-PCR and hydroxyproline method.Results HEK293 cells were successfully transfected with the constructed recombinant pMSCV/sIL-13 Rα2 and expressed sIL-13 Rα2.Type Ⅰ collagen synthesis was promoted by 50 μg·L-1 IL-13 in scar fibroblast CRL-1762 cells.But the promoting effect of IL-13 was inhibited by 50 μmol·L-1 sIL-13 Rα2.Conclusion Type Ⅰ collagen synthesis can be inhibited by targeting IL-13 with sIL-13 Rα2.in fibroblasts.