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目的:观察右归丸含药血清对骨髓基质干细胞的骨向分化诱导作用,通过右归丸含药血清对骨髓基质干细胞Akt、mTOR磷酸化水平的影响,探讨右归丸含药血清促骨髓基质干细胞骨向分化的部分作用机制。方法:制备高中低3个剂量的右归丸大鼠含药血清,提取大鼠骨髓基质干细胞,传至第3代,将细胞随机分为5组:空白对照组、空白血清组、低剂量含药血清组、中剂量含药血清组、高剂量含药血清组。分别给予不同浓度血清及PBS干预24 h后,检测各组细胞中碱性磷酸酶活性、骨钙素表达水平及Akt、mTOR磷酸化水平。结果:空白对照组与空白血清组之间比较,ALP、Osteocalcin、p-Akt、p-mTOR表达水平无统计学差异(P>0.05),与空白对照组和空白血清组比较,3个剂量含药血清组细胞中ALP活性及Osteocalcin、p-Akt、p-mTOR表达水平均不同程度提高(P>0.01),但中剂量和高剂量含药血清组ALP活性及Osteocalcin、p-Akt、p-mTOR表达水平均无显著性差异(P>0.05)。结论:右归丸含药血清具有一定的促进骨髓基质干细胞骨向分化的作用,该作用可能是通过提高Akt、mTOR的磷酸化水平而实现的。
OBJECTIVE: To observe the effect of Youguiwan containing serum on bone marrow stromal differentiation induced by bone marrow stromal cells, and to study the effect of Youguiwan containing serum on the phosphorylation of Akt and mTOR in bone marrow stromal cells Stem cells to differentiate part of the mechanism of action. Methods: Three doses of high, medium and low dose of Yougui Wan containing serum were prepared and the rat bone marrow stromal cells were extracted and passaged to the third passage. The cells were randomly divided into five groups: blank control group, blank serum group, low dose drug Serum group, middle dose serum group and high dose serum group. After being treated with different concentrations of serum and PBS for 24 hours, the activity of alkaline phosphatase, osteocalcin, phosphorylation of Akt and mTOR in each group were detected. Results: There was no significant difference in the expression of ALP, Osteocalcin, p-Akt and p-mTOR between blank control group and blank serum group (P> 0.05). Compared with blank control group and blank serum group, The ALP activity and the expression of Osteocalcin, p-Akt and p-mTOR in the serum of the drug-treated group were all increased to some extent (P> 0.01) mTOR expression levels were no significant difference (P> 0.05). Conclusion: Yougui pill containing serum can promote the bone marrow stromal differentiation to a certain extent, and this effect may be achieved by increasing the phosphorylation level of Akt and mTOR.