摘要
目的观察地塞米松(Dex)和1,25(OH)2D3(D3)对骨髓基质细胞(MSCs)成骨及成脂分化的影响。方法以离心法分离培养人MSCs,以10-7mol/L Dex和/或10-8mol/L1,25(OH)2D3作为分化诱导剂对细胞进行干预,分别用细胞碱性磷酸酶(ALP)染液试剂盒及苏丹Ⅲ染液对成骨细胞和脂肪细胞进行组织化学染色,计数;使用RT-PCR技术在转录水平检测成骨细胞标记物骨桥蛋白(OPN)及脂肪细胞标记物过氧化酶体增殖激活受体γ2(PPARγ2)mRNA的表达。结果细胞染色结果表明各干预组ALP+细胞百分比均较对照组增加,与对照组相比有显著性差异(P<0·05),苏丹Ⅲ+细胞百分比Dex组较对照组增多,D3组较对照组减少,差异有显著性(P<0·05),Dex+D3组较Dex组苏丹Ⅲ+细胞数明显减少,两者相比差异有显著性(P<0·05);OPN mRNA及PPARγ2mRNA表达未在对照组测得,Dex诱导了OPN mRNA及PPARγ2mRNA表达,1,25(OH)2D3诱导OPN mRNA表达,并抑制Dex诱导的PPARγ2mRNA的表达。结论Dex促进MSCs的成骨分化及成脂分化,1,25(OH)2D3促进MSCs的成骨分化的同时抑制其成脂分化,与Dex合用抑制Dex成脂分化作用,强化了其成骨分化作用,反映了成骨细胞与脂肪细胞间存在的反变关系,表明两者来源于同一前体细胞的可能性。
Objective To observe the effects of dexamethasone （DEX） and 1,25（ OH）2 Ds on osteogenic and adipocytic differentiation of human marrow stromal cells （MSCs） in vitro. Methods The primary human MSCs were isolated and cuhured. DEX （ 10 ^- 7 mol/L） and/or 1,25 （OH） 21）3 （ 10^ -8 mol/L ） were added as differentiation inducers. The histochemical staining was performed with ALP staining to identify osteogenic cells and Sudan HI to identify adipocytic cells. The numbers of positive cells were counted. The expression of osteogenic marker osteopontin （OPN） and the adipocytic marker PPARγ2 mRNA was examined by reverse transcriptase polymerase chain reaction （RT-PCR）. Results The percentages of ALP-positive cells and Sudan Ⅲ -positive cells in cultures with DEX were significantly higher than that of control （P 〈 0.05）. For those cultured with 1,25（OH）2D3 , the percentage of ALP-positive cells was higher （ P 〈 0.05） ,while that of Sudan m -positive cells was lower than that of control （P 〈 0.05）. The expression of OPN mRNA and PPARγ 2 mRNA was induced by DEX and 1,25（OH）2D3 promoted the expression of OPN mRNA but inhibited the stimulatory effect of DEX on PPARγ2 mRNA. Conclusions DEX induces human MSCs differentiation into osteoblasts and adipocytes and 1,25（OH）2D3 induces human MSCs differentiation into osteoblasts but inhibit adipocytic differentiation. The inverse relationship between the differentiation of osteoblast and adipocyte in human MSCs suggesting the possibility of a common precursor of these two kinds of cells.
出处
《中国骨质疏松杂志》
CAS
CSCD
2006年第5期447-451,共5页
Chinese Journal of Osteoporosis
关键词
地塞米松
1
25(OH)2D3
人骨髓基质细胞
成骨分化
成脂分化
Dexamethasone
1,25（OH）2D3
Human marrow stromal cells
Osteogenic differentiation
Adipocytic differentiation
