异补骨脂素促进大鼠骨髓间充质干细胞向成骨细胞分化并抑制其向脂肪细胞分化

Isopsoralen stimulate osteogenesis and inhibit adipogenesis of bone marrow mesenchymal stem cells

目的:研究异补骨脂素是否影响骨髓间充质干细胞向成骨细胞和脂肪细胞分化的能力。方法:不同浓度异补骨脂素分别处理在成骨或成脂诱导培养液中培养的大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BM-MSCs),培养14天,分别行亚甲基蓝、碱性磷酸酶(alkaline phosphatase,ALP)和油红O染色,并用提取处理后细胞总RNA和蛋白,RT-PCR和West-ern blot检测成骨或成脂相关基因的表达差异。结果:①异补骨脂素可提高大鼠BM-MSCs的总成纤维细胞集落形成单位(CFU-f)及ALP阳性CFU-f形成效率,并促进成骨相关基因的表达;②异补骨脂素可抑制大鼠BM-MSCs诱导脂肪滴的形成及过氧化物酶增殖体激活受体(peroxisome proliferator-activated receptor-γ,PPARγ)基因和蛋白的表达。结论:异补骨脂素可促进大鼠BM-MSC向成骨细胞分化并抑制其向脂肪细胞分化,其作用效果较补骨脂素更强。

Objective：To determine the effect of isopsoralen on the osteogensis and adipogenesis of bone marrow mesenchymal stem cells（BM-MSCs）.Methods：Rat BM-MSCs were cultured in the absence or presence of isopsoralen for 14 days,and resulting cells were stained with methylene blue,Oil Red O and detected for alkaline phosphatase（ALP）.RNA and protein were isolated from cells,and expression levels of osteogenesis and adipogenesis related genes were evaluated by RT-PCR and Western blot.Results：①The efficiency of CFU-f and ALP positive CFU-f formation by BM-MSCs was enhanced,and the expressions of osteogenesis related genes were up-regulated significantly in isopsoralen-treated cultures compared with control cultures.②The adipocyte forming efficiency and the gene and protein expressions of peroxisome proliferator-activated receptor-γ（PPARγ） were inhibited significantly in isopsoralen-treated cultures compared with control cultures.Conclusion：Our results indicate that isopsoralen can stimulate osteogensis and inhibit adipogenesis of BM-MSCs,and the stimulating effect of isopsoralen is better than psoralen.