人可溶型IL-13受体α#23545;成纤维细胞胶原生成作用的研究

项目名称： 人可溶型IL-13受体α#23545;成纤维细胞胶原生成作用的研究

项目编号： No.30860118

项目类型： 地区科学基金项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 石小玉

作者单位： 南昌大学

项目金额： 24.1万元

中文摘要： 从人胶质瘤细胞系U251克隆IL-13 Rα#65292;把IL-13RαcDNA连接在表达载体pMSCV, 在其cDNA的N端插入分泌肽序列（81bp），分子量（0.2×6 D）滤膜离心浓缩培养液,亲和层析和SDS-PAGE 等实验结果表明构建pMSCV/sIL-13Rα#36733;体。pMSCV/sIL-13Rα#36716;染入HEK293细胞，Western blot 实验结果显示构建表达sIL-13Rα#30340;细胞模型。RT-PCR和羟脯氨酸法实验结果表明50μl/L sIL-13Rα#33021;有效抑制IL-13刺激人疤痕成纤维细胞系CRL-1762和人肝成纤维细胞系LI90合成胶原。小鼠腹腔注射硫代乙酰胺构建人肝纤维化裸小鼠模型，小鼠皮下种植人疤痕成纤维细胞系CRL-1762构建人皮肤疤痕裸小鼠模型。 静脉注射1ng-200ngIL-13/100g小鼠体重,并静脉注射1ng-300ng sIL-13 受α100g小鼠体重, 免疫组织化学实验结果表明sIL-13Rα#26377;效抑制肝纤维化和皮下疤痕生长。在实验中，还观察到LPA、EGF和TNF-α33021;诱导IL-13Rα#34920;达并抑制胶原蛋白合成。

中文关键词： sIL-13 Rα#65307;胶原；成纤维细胞；纤维化

英文摘要： IL-13 R α was cloned from human glioma cell line U251. IL-13 R α cDNA was connected in pMSCV vector. 81 bp of secretion peptide sequence were inserted in IL-13 R α cDNA N end. The membrane of molecular weight (0.2 x 106 D) filtered concentrated medium. The experimental results of affinity chromatography and SDS-PAGE showed that pMSCV/sIL-13 R α vector was constructed. HEK293 was infected by pMSCV/sIL-13Rα The results of Western blot test indicated that cells model of expressing sIL-13 R α was constructed. The results of RT-PCR and hydroxyproline method showed that 50 μol /L sIL-13 Rα can effectively restrain collagen synthesis in human scar fibroblast CRL-1762 and human liver fibroblast LI90 stimulated by IL-13. The nude mice model of human liver fibrosis was constructed by intraperitoneal injection of thioacetamide. The mice were given subcutaneous injection of human scar cell lines CRL-1762 to construct the scar model. The mice of models were treated with 1ng-200 ng IL-13/100g mouse weight and 1ng-300 ng sIL-13 Rα100g mouse weight through intravenous injection. Immunohistochemical experimental results showed that sIL-13 α effectively inhibited liver fibrosis and subcutaneous scar growth in mice of model. In the experiments, we also observed that LPA, EGF and TNF alpha can induced IL-13 R α expression and inhibited collagen protein synthesis.

英文关键词： sIL-13 Rα#65307;collagen; fibroblast; fibrosis