摘要
目的利用大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)建造体外实验模型,探讨间歇应用甲状旁腺激素1-34(parathyroid hormone 1-34,PTH 1-34)对BMSCs成脂分化中脂蛋白脂肪酶(lipoprotein lipase,LPL)和碱性磷酸酶(alkaline phosphatase,ALP)的影响。方法 (1)全骨髓贴壁培养法分离出BMSCs,体外培养与鉴定;(2)以P3代BMSCs为体外实验模型,分为两组:单纯诱导液组和PTH组。单纯诱导液组:BMSCs+DMEM培养基+诱导液;PTH组:BMSCs+DMEM培养基+诱导液+PTH 1-34;(3)采用间歇方式将不同浓度的PTH 1-34加入PTH组的BMSCs培养基,14天后采用ELISA法测定LPL活性,采用RT-PCR法测定ALP的活性。结果 (1)P3代细胞表达抗原CD29、CD44,不表达抗原CD45,基本符合BMSCs表面标志物特征;(2)成脂诱导液诱导BMSCs分化21天后,油红O染色可见胞内脂滴出现,符合脂肪细胞的特性;(3)间歇应用不同浓度的PTH作用于BMSCs后,与对照组比较,LPL的表达量下降,ALP活性增加,并呈剂量依赖性。结论小剂量间歇应用PTH 1-34可使BMSCs的成脂分化中LPL表达量降低,ALP表达量增加,证明小剂量PTH 1-34可抑制BMSCs成脂分化,促进其成骨分化。
Objective To establish rat bonemarrowmesenchymal stem cell experimental model,and to exploretheeffect of intermittent application of parathyroid hormoneon lipoprotein lipase（ LPL） and alkalinephosphatase（ ALP） during adipogenic differentiation by BMSCs. Methods（ 1） BMCSs wereseparated and expanded by wholebonemarrowadherent culturemethod,and they wereidentified in vitro.（ 2） TheP3 of BMSCs was taken for thetest in vitro. They weredivided into 2 groups： pureinduced liquid group（ BMSCs ＋ induced liquid ＋ DMEMmedium） and PTH group（ BMSCs ＋ induced liquid ＋ DMEMmedium ＋PTH1-34）.（ 3） Different concentrations of PTH1-34 wereintermittently additioned to BMSCs medium. After 14 days LPL activity was measured using ELISA. Expression of ALP mRNA was measured using real-timePCR. Results TheP3 cells expressed CD44 and CD29,but did not express CD45,which wereconsistent of BMSCs characteristics. By culturing in adipogenic medium for 21 days,Oil Red O staining could beseen,indicating that thecells weresuccessfully induced to adipocytes. Compared with thecontrol group,mRNA expression of ALP decreased but LPL activity increased after intermittent useof different concentrations of PTH1-34. Theeffect was dose-dependent. Conclusion Intermittent application in lowdoses of PTH1-34 decreaseLPL activity but increasemRNA expression of ALP during adipogenic differentiation by BMSCs,indicating that lowdoses of PTH1-34 inhibit adipogenic differentiation but promoteosteogenic differentiation.
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2015年第12期1460-1464,共5页
Chinese Journal of Osteoporosis
基金
太原市社会发展科技项目(11027604)
关键词
甲状旁腺激素1-34
骨髓间充质干细胞
成脂分化
脂蛋白脂肪酶
碱性磷酸酶
Parathyroid hormone 1-34
Bone marrow mesenchymal stem cells
Adipogenic differentiation
Lipoprotein lipase
Alkaline phosphatase
