脂肪细胞条件培养基对成骨前体细胞成骨分化能力的影响

Effect of fat-cell conditioned-medium on differentiation of MC3T3-E1

目的探讨脂肪细胞条件培养基对成骨前体细胞(MC3T3-E1)成骨方向分化能力的影响。方法制备脂肪细胞条件培养基(FCCM)培养3T3-E1,设常规培养液为对照培养基组(CM)。分别在培养7、14d后行Real-time PCR和West-ern-blot检测成骨相关转录因子Runx2(Runt-related transcription factor 2)、碱性磷酸酶(ALP)、骨钙素(bone gamma-carbox-yglutamic-acid-containing protein,BGP),并在第14天时行ALP活性检测、ALP染色和茜素红染色。结果 FCCM组较对照组的3T3-E1的Runx2、ALP、BGP基因下调;同时Runx2、ALP、BGP蛋白水平下调;且FCCM组较对照组ALP活性降低,ALP染色和茜素红染色结果均为FCCM组为阴性,对照组阳性。结论 FCCM可下调成骨前体细胞成骨方向分化能力。

Objective To investgate the influence of fat-cell conditioned-medium（FCCM） on the differentiation of MC3T3-E1.Methods FCCM and control medium（CM） were prepared to culture MC3T2-E1 cells.The expression of Runt-related transcription factor 2（Runx2）,alkaline phosphatase（ALP）,and bone gamma-carboxyglutamic-acid-containing protein（BGP） was detected by using real-time PCR and Western blotting.ALP activity was determined by pnitro phenyl phosphate assay.Calcium nodules were examined by using Van Gieson and alizarin red method.Results The expression of Runx2,ALP and BGP,and activity of ALP in FCCM group were reduced as compared with CM group.ALP staining and Alizarin red staining in FCCM group was weaker than CM group.Conclusion FCCM can down-regulated the osteogenic differentiation of MC3T3-E1.