抗波形纤维蛋白单抗对大鼠前体脂肪细胞增殖分化及波形纤维形态的影响  

Effects of cytoplasm microinjection of anti-vimentin monoclonal antibody on the proliferation and differentiation of rat preadipocytes and morphology of vimentin

作　　者：田志华[1] 杨公社[1] 赵兴波[2] 何大澄[3] 

机构地区：[1]西北农林科技大学畜牧兽医学院,陕西杨凌712100 [2]中国农业大学动物科技学院,北京100094 [3]北京师范大学生命科学学院,北京100875

出　　处：《动物学报》2003年第6期807-812,共6页ACTA ZOOLOGICA SINICA

基　　金：国家自然科学基金项目 (No .3 93 70 5 0 3 );国家高技术研究发展计划 (863计划 )(No .2 0 0 1AA2 13 12 1)资助~~

摘　　要：将抗波形纤维蛋白单抗通过显微注射导入未分化的大鼠前体脂肪细胞的细胞质 ,观察其对细胞增殖、分化 ,及其中等纤维———内源波形纤维形态的影响。以注射人IgG和未注射的前体脂肪细胞作为对照 ,发现注射单抗的前体脂肪细胞呈现以下特征 :( 1)细胞呈典型的凋亡形态 ,经Hoechst 33342着染可见胞核呈致密浓染的颗粒状或块状荧光 ,胞内脂滴几乎没有 ;( 2 )增殖的细胞数和甘油三酯值均极显著低于对照组 ;( 3)波形纤维蛋白间接免疫荧光染色微弱 ,纤维束解聚成短小团块状 ,分布无序、弥散。上述结果表明 ,抗波形纤维蛋白单抗对前体脂肪细胞的增殖和分化有明显阻滞作用。Intermediate filaments (IFs) are prominent structural feature of the cytoskeleton of higher eukaryotic cells. A variety of higher vertebrate cells of mesenchymal origin, such as fat tissue, have been shown to contain vimentin IF. The specific expression of vimentin IFs was maintained during adipose conversion. A better understanding of the significance of the disrupution of vimentin becomes important in characterizing the mechanism through which vimentin affects fat generation. The present experiments were carried out to examine the effects of anti vimentin monoclonal antibody microinjected into cytoplasm on the proliferation and differentiation of rat preadiocyte and morphology of endogenous vimentin. Preadipocytes were isolated from adipose tissue of Sprague Dawley rats, aged 20 days,by collagenase digestion. Cells were seeded at a density of 50-100 cells on each glass coverslip in M199 medium plus 10% fetal calf serum and cultured at 37 ℃ under a humidified atmosphere of 95% air∶5% CO 2. One day later, at the initiation of the differentiation, purified anti vimentin McAb was microinjected into the cells,at a concentration of 1 mg/ml, while setting cells of both human IgG microinjection and non microinjection as control groups. To determine if disruption of the vimentin IF network would affect the capacity of predipocytes to accumulate lipid droplets during adipose conversion, the marker enzyme triglyceride (TG), as a specific and quantifiable marker for adipose conversion, was assayed spectrophotometrically. Finally, at the indicated times following microinjection, the cells were processed for anti vimentin indirect immunofluorescence, to investigate the morphology alternation of the vimentin intermediate filament. The following characteristics of cells with anti vimentin McAb microinjection were oberserved: (1) the cells were under apoptosis condition, hardly without lipid droplet in the cytoplasm, and nuclear fragmented into blue condensed particles or block detecting by Hoechst33342; (2) the t

关 键 词：显微注射 抗波形纤维蛋白单抗 大鼠前体脂肪细胞 波形纤维 大鼠 肥胖 高血脂症 

分 类 号：S852.2[农业科学—基础兽医学]