骨形成蛋白7对大鼠不同部位脂肪源干细胞分化成棕色样脂肪细胞的影响

Effect of Bone Morphogenetic Protein 7 on Differentiation of Adipose Derived Mesenchymal Stem Cells into Brown Adipocytes in Rats

目的探讨骨形成蛋白7对大鼠不同部位脂肪组织脂肪源间充质干细胞(AD-MSCs)诱导分化成棕色样脂肪细胞的影响。方法分离大鼠背部肩胛间区棕色脂肪组织(i BAT)、双侧下肢腹股沟皮下白色脂肪组织(s WAT)和附睾白色脂肪组织(e WAT),分别提取脂肪源间充质干细胞培养,用骨形成蛋白7诱导干细胞分化。细胞免疫荧光染色、油红O染色鉴定棕色样脂肪细胞,PCR法检测棕色脂肪相关基因表达。结果 i BAT和s WAT组诱导细胞中出现成簇聚集的空泡状细胞,空泡状细胞油红O染色为红色,解偶联蛋白1免疫荧光染色阳性。e WAT组仅有散在的空泡状细胞,解偶联蛋白1免疫荧光染色阴性。反转录PCR显示i BAT和s WAT组细胞高表达解偶联蛋白1基因,未检测到e WAT组细胞表达解偶联蛋白1基因。结论大鼠体内不同部位脂肪组织源干细胞的基因表达及分化潜能不同。

Objective To evaluate the effect of bone morphogenetic protein （BMP7） on the differentiation of adipose derived mesenchymal stem cells （AD-MSCs） isolated from different adipose tissues into brown adipocytes in rats.Methods Primary AD-MSCs were isolated from rate interscapular brown adipose tissue （iBAT）,inguinal subcutaneous white adipose tissue （sWAT）,and epididymal white adipose tissue （eWAT）,respectively,and then cultivated in vitro.Differentiation of AD-MSCs into brown adipocytes was induced by BMP7.The characteristics of brown adipocytes were detected by immunofluorescence staining and oil red staining of cells.The expression levels of brown adipocyte-related genes were detected by polymerase chain reaction.Results AD-MSCs from iBAT and sWAT were differentiated into cluster multilocular cells,which were stained red by oil red ＂O＂ staining and showed uncoupling protein 1-positive by immunofluorescent staining method.AD-MSCs from eWAT had a small number of scattered multilocular cells and showed uncoupling protein 1-negative.The resu1ts of reverse transcription-polymerase chain reaction showed that the uncoupling protein 1 gene was highly expressed in the iBAT group and sWAT group but was negative in the eWAT group.Conclusion AD-MSCs isolated from different adipose tissues in rats have different gene expression profiles and differentiation potentials.