HIF-1α转染骨骼肌细胞的诱导产物VEGF对血管内皮细胞增殖的影响  

HIF-1α gene transfected into skeletal muscle cells and the inducible VEGF effect on the proliferation of vascular endothelium cells

作　　者：陈博[1] 陈剑秋[1] 

机构地区：[1]天津医科大学第二医院普通外科,天津300211

出　　处：《天津医科大学学报》2010年第3期420-423,共4页Journal of Tianjin Medical University

摘　　要：目的:将缺氧诱导因子-1α(HIF-1α)基因转染体外培养大鼠骨骼肌细胞,观察转染后细胞上清液中VEGF蛋白的表达及其对血管内皮细胞增殖的影响。方法:利用机械和酶消化法分离培养大鼠骨骼肌细胞,贴块法培养血管内皮细胞;用阳离子脂质体介导pHOX/HIF-1α质粒转染骨骼肌细胞,ELISA法检测转染后VEGF蛋白的分泌情况,MTT法检测其对血管内皮细胞增殖的影响。结果:成功将HIF-1α基因转染大鼠骨骼肌细胞,转染组细胞培养上清中244、8、72和96 h检测到VEGF含量分别为(1 308.40±102.00)(、1 449.20±69.86)(、1 802.30±96.71)和(1 494.95±86.24)pg/ml,明显高于对照组;转染后骨骼肌细胞分泌的VEGF对血管内皮细胞有显著的促增殖作用。结论:HIF-1α基因转染大鼠的骨骼肌细胞后可增加具有正常功能的VEGF蛋白的分泌,促进血管内皮细胞的增殖。Objective： To study the effect of the VEGF protein in skeletal muscle cells of rat culture medium on the proliferation of primary culture endothelium cells,which were transfected by HIF-1α gene in vitro. Methods： Skeletal muscle cells and vascular endothelium cells were obtained from the Wistar rats. pHOX/HIF-1α was transfected into primary culture skeletal muscle cells with cationic liposome （LipofectamineTM 2000）.The secreted VEGF protein in the supernatant after transfection was demonstrated by ELISA,it＇ s effect on the proliferation of vascular endothelium cells were evaluated through MTT. Results： HIF-1α was transfected into primary culture skeletal muscle cells in vitro successfully, the VEGF protein in culture medium of skeletal muscle cells after transfection 24,48,72 and 96 h were （1 308.40±102.00）, （1 449.20±69.86）, （1 802.30±96.71） and （1 494.95±86.24） pg/ml respectively. Furthermore, the cells secreted VEGF protein had the ability to stimulate the proliferation of vascular endothelium cells. Conclusion： HIF-IcL is transfected into primary culture skeletal muscle cells successfully and the secreted VEGF protein can promote the proliferation of primary culture vascular endothelium cells.

关 键 词：缺氧诱导因子-1Α 骨骼肌细胞 内皮细胞 血管内皮生长因子 基因转染 

分 类 号：Q7[生物学—分子生物学] Q813