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| 甲状旁腺激素对大鼠骨髓间充质干细胞成脂分化中LPL和ALP的影响 |
| Effect of parathyroid hormone on LPL and ALP during adipogenic differentiation by rat bone marrow mesenchymal stem cells |
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| DOI：10.3969/j.issn.1006.7108.2015.12.008 |
| 中文关键词: 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 |
| 英文关键词:Parathyroid hormone 1-34 Bone marrow mesenchymal stem cells Adipogenic differentiation Lipoprotein lipase Alkaline phosphatase |
| 基金项目:太原市社会发展科技项目(11027604) |
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| 中文摘要: |
| 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加人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。 |
| 英文摘要: |
| Objective To establish rat bone marrow mesenchymal stem celt experimental model, and to explore the effect of intermittent application of parathyroid hormone on lipoprotein lipase ( LPL) and alkaline phosphatase ( ALP) during adipogenic differentiation by BMSCs. Methods (1) BMCSs were separated and expanded by whole bone marrow adherent culture method, and they were identified in vitro. (2) The P3 of BMSCs was taken for the test in vitro. They were divided into 2 groups； pure induced liquid group (BMSCs + induced liquid + DMEM medium) and PTH group ( BMSCs + induced liquid + DMEM medium + PTH 1-34). (3) Different concentrations of PTH 1-34 were intermittently additioned to BMSCs medium. After 14 days LPL activity was measured using ELISA. Expression of ALP mRNA was measured using real-time PCR. Results The P3 cells expressed CD44 and CD29, but did not express CD45, which were consistent of BMSCs characteristics. By culturing in adipogenic medium for 21 days, Oil Red O staining could be seen, indicating that the cells were successfully induced to adipocytes. Compared with the control group, mRNA expression of ALP decreased but LPL activity increased after intermittent use of different concentrations of PTH 1-34. The effect was dose-dependent. Conclusion Intermittent application in low doses of PTH 1-34 decrease LPL activity but increase mRNA expression of ALP during adipogenic differentiation by BMSCs, indicating that low doses of PTH1 -34 inhibit adipogenic differentiation but promote osteogenic differentiation. |
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