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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。

英文摘要:

Objective: To investigate the effect of euphornin, an active component from Euphorbia Helioscopia L, on the osteogenic differentiation in rat mesenchymal stem cells (rMSC). Methods: rMSC were isolated from rat femurs, cultured and induced into osteoblasts. MTT assay was applied to detect the role of euphornin on cell proliferation. Alkaline phosphatase (ALP) assay and calcium content measurement were assigned to quantify the suppressed osteogenesis and alizarin reds (ARS) staining was conducted to visualize it. Quantitative real-time PCR (Q-PCR) was used to evaluate the mRNA expression of marker genes in osteogenesis and master regulators in BMP pathway. Results: Euphornin suppressed osteogenesis of rMSC in a dose-dependent manner, and suppressed its proliferation to some extent. In euphornin-treated cells, the expression of typeⅠ collagen (COL-Ⅰ) and osteopontin (OPN) were down-regulated at mRNA level on Day 4 and 8 respectively. The mRNA expression of bone morphogenetic protein-2 (BMP2), Runt-related transcription factor-2 (Runx2), and osterix (Osx) also decreased. Conclusions: Euphornin suppresses the osteogenic differentiation of rMSC. The suppression role might be exerted by down-regulating the expression of key transcriptional factors in BMP pathway.

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