摘要
目的观察不同糖浓度下吡格列酮对大鼠骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)向脂肪细胞分化的影响,探讨葡萄糖和吡格列酮对骨代谢的影响。方法采用体外细胞培养技术自大鼠股骨和胫骨中分离BMSCs进行纯化、培养,在诱导成脂培养基(地塞米松、3-异丁基-1-甲基黄嘌呤(IBMX)、胰岛素)中诱导BMSCs分化为脂肪细胞,实验分为高糖组(葡萄糖浓度为50mmol.L-1)及正糖组(葡萄糖浓度为25mmol.L-1),两组分别用不同浓度的吡格列酮(0、0.1、1μg.mL-1)干预分化过程各21d,油红O(Oil Red O)染色鉴定分化后的脂肪细胞,光镜下观察橙红色脂滴沉着的细胞比例。实时荧光定量PCR测定脂肪细胞特异性标志LPL、PPARγmRNA的表达。结果诱导分化培养21d后,油红O染色结果显示随糖浓度增加脂肪细胞数量增多,PCR结果显示HG+NC组比NG+NC组LPL、PPARγmRNA表达分别增加1.40倍(P<0.05)和1.63倍(P<0.05),在两种糖浓度下,分别加入吡格列酮后脂肪细胞分化均显著增加,数量明显增多,体积明显增大。与NG+NC组相比,NG+LP组LPL和PPARγmRNA表达分别增加1.43倍(P<0.05)和1.50倍(P<0.05),NG+GP组mRNA水平增加更明显,HG+LP组和HG+HP与HG+NC组相比,LPL和PPARγmRNA表达增加更明显。结论高糖会促进BMSCs向脂肪细胞分化,可能为糖尿病性骨质疏松形成的重要机制。吡格列酮有显著增加BMSCs向脂肪细胞方向分化的作用,且随药物剂量的增加,其诱导成脂分化的效应越明显。吡格列酮可能通过诱导BMSCs向脂肪细胞分化增多而向成骨细胞分化减少从而导致成骨作用减弱,这可能是吡格列酮致骨质疏松的重要机制。
Objective To examine the effect of pioglitazone on rat bone mesenchymal stem cells （BMSCs） differentiation into adipocytes in different glucose concentrations and to investigate the modulation of glucose and pioglitazone on bone metabolism. Methods BMSCs were harvested from the femurs and tibias bones of the rat, then separated, purified, proliferated and treated with an adipogenic medium（including dexamethasone, 3-isobutyl- 1-methyl-xanthine, insulin ）, using different doses of pioglitazone （0, 0.1, 1μg· mL^-1 ） to intervene the adipogenic differentiation in high glucose group（50 mmol· L^-1 ） and normal glucose group（25 mmol· L^-1 ） for 21 days, respectively. Differentiated adipocytes were identificated by Oil Red O and the expression of adipose-specific mRNAs： LPL and PPAR ）＇ were assayed by real time PCR. Results After adipogenic had been induced for 21 days, the number of adipocyes detected by Oil Red O increased in high glucose group, the mRNAs expression of LPL and PPAR γ increased 1.40 and 1.63 folds in high glucose group compared with normal glucose group. Addition of pioglitazone dramatically promoted the differentiation of adipocytes in a dose-dependent manner in both concentrations of glucose. Compared with NG ＋ NC group, the mRNA expression of LPL and PPAR γ grew 1.43 and 1.50 folds in NG ＋ LP group and 2.4l and 2.17 folds in NG ＋ GP group respectively. The effect of pioglitazone on adipogenesis in high glucose group exceeded normal glucose group. Conclusion High concentration glucose promoted the differentiation of BMSCs into adipocytes. This ohservation provides a potential mechanisms of diabetes-related osteoporosis. Pioglitazone dramatically increased BMSCs differentiating into adipocytes and this effect shows dose-dependent augmentation. Pioglitazone decreases bone mass by promoting adipogenesis and inhibiting osteoblastic diffenrentiation and it may be the important potential pathogenisis of osteoporosis caused by pioglitazone.
出处
《中国骨质疏松杂志》
CAS
CSCD
2008年第8期561-566,共6页
Chinese Journal of Osteoporosis
