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1、动物学报 49(6):807～812,2003ActaZoologicaSinica抗波形纤维蛋白单抗对大鼠前体脂肪细胞增殖3分化及波形纤维形态的影响①③①33②③田志华 杨公社 赵兴波 何大澄(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)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
2、,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Hoechst33342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。同时为揭示波形纤维蛋白可能参与了脂肪细胞形成这一重要生理过程提供了证据[动物学报49(6):807～812,2003]。关键词 显微注射 抗波形纤维蛋白单抗 大鼠前体脂肪细胞
3、 波形纤维Effectsofcytoplasmmicroinjectionofanti2vimentinmonoclonalantibodyonthe3proliferationanddifferentiationofratpreadipocytesandmorphologyofvimentin①③①33②③TIANZhi2HuaYANGGong2SheZHAOXing2BoHEDa2Cheng(①CollegeofHusbandryandVeterinary,UniversityofNorthwesternAgro2Fores
4、trySci2technology,Yangling712100,Shaanxi,China)(②CollegeofAnimalSci2techenology,ChinaAgricultureUniversity,Beijing100094,China)(③CollegeofLifeSciences,BeijingNormalUniversity,Beijing100875,China)AbstractIntermediatefilaments(IFs)areprominentstructuralfeatureofthecyto
5、skeletonofhighereukaryoticcells.Avarietyofhighervertebratecellsofmesenchymalorigin,suchasfattissue,havebeenshowntocontainvimentinIF.ThespecificexpressionofvimentinIFswasmaintainedduringadiposeconversion.Abetterunderstandingofthesignificanceofthedisruputionofvimentinb
6、ecomesimportantincharacterizingthemechanismthroughwhichvimentinaffectsfatgenera2tion.Thepresentexperimentswerecarriedouttoexaminetheeffectsofanti2vimentinmonoclonalantibodymicroinjectedintocytoplasmontheproliferationanddifferentiationofratpreadiocyteandmorphologyofen
7、dogenousvimentin.PreadipocyteswereisolatedfromadiposetissueofSprague2Dawleyrats,aged20days,bycollagenasedigestion.Cellswereseededatadensityof50-100cellsoneachglasscoverslipinM199mediumplus10%fetalcalfserumandculturedat37℃underahumidifiedatmosphereof95%air∶5%CO2.Oneda
8、ylater,attheinitiationofthedifferentiation,puri2fiedanti2vimentinMcAbwasmicroinjectedintothecells,ataconcentrationof1mg/ml,whileset
