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目的:观察泽漆主要活性成分大戟苷(euphornin)对大鼠骨髓间充质干细胞(rMSC)成骨分化的影响。方法:从大鼠股骨中分离培养rMSC,并诱导其成骨分化。用MTT法检测细胞增殖情况,通过茜素红染色,碱性磷酸酶(ALP)活性检测和钙含量测定分别定性、定量地判断其在成骨分化中的效果。实时定量PCR(Q-PCR)检测主要成骨标志因子骨桥蛋白(OPN)和一型胶原蛋白(COL-Ⅰ)及主要转录因子骨形成蛋白2(BMP2)、Runt相关转录因子2(Runx2)和Osterix(Osx)mRNA的表达。结果:大戟苷能剂量依赖性地抑制rMSC成骨分化,并一定程度地抑制其细胞增殖。COL-Ⅰ和OPN的表达在第4、8天分别有显著下降。BMP2、Runx2和Osx等关键转录因子的表达也被明显抑制。结论:大戟苷能抑制rMSC成骨分化,其作用主要是通过抑制BMP通路相关因子的表达而实现的。
OBJECTIVE: To observe the effect of euphornin, the main active component of Ze Ze, on osteogenic differentiation of rat bone marrow mesenchymal stem cells (rMSC). Methods: The rMSC was isolated and cultured from rat femur, and its osteogenic differentiation was induced. Cell proliferation was detected by MTT assay. The effect of osteogenic differentiation was qualitatively and quantitatively determined by alizarin red staining, alkaline phosphatase (ALP) activity assay and calcium determination. The major osteogenic markers osteopontin (OPN) and collagen type Ⅰ (COL-Ⅰ) as well as the major transcription factors BMP2, Runt-related transcription factor 2 (Runx2) were detected by real- And Osterix (Osx) mRNA expression. Results: Anthradin inhibited the osteogenic differentiation of rMSCs in a dose - dependent manner and inhibited its proliferation to a certain extent. The expression of COL-I and OPN were significantly decreased on the 4th and 8th day respectively. The expression of key transcription factors such as BMP2, Runx2 and Osx was also significantly inhibited. Conclusion: Euphoridin can inhibit the osteogenic differentiation of rMSCs, and its effect is mainly through inhibiting the expression of BMP pathway related factors.