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目的:观察健脾补肾方含药血清对体外培养的兔骨髓间充质干细胞(rMSCs)定向分化的影响,揭示本方临床应用防治激素性股骨头坏死的药理机制。方法:以健脾补肾方含药血清作用于体外培养的rMSCs,通过对碱性磷酸酶(ALP)活性的的动态检测结果和四环素荧光标记的矿化结节的出现数量作为分析各组诱导液能否诱导rMSCs分化为成骨细胞以及促进骨向分化强度的指标。结果:rMSCs培养的第12天,健脾补肾组ALP活性高于空白对照组(P<0.05);从第8天之后,健脾补肾加成骨组ALP活性高于成骨组(P<0.05);更换相应培养液22d后,其余3组形成的钙结节数量明显多于空白对照组(P<0.01);健脾补肾加成骨组钙结节数量多于成骨组(P<0.05)。结论:健脾补肾方含药血清可以促进rMSCs的骨向分化,且与经典的成骨诱导剂合用具有协同作用。这可能揭示了临床应用该方防治激素性股骨头坏死(SINFH)的细胞学机制。
OBJECTIVE: To observe the effect of Jianpi Bushen recipe-containing serum on the directional differentiation of cultured rabbit bone marrow mesenchymal stem cells (rMSCs) in vitro, and to reveal the clinical pharmacological mechanism of prevention and treatment of steroid-induced femoral head necrosis. Methods: The serum containing Jianpi Bushenfang was used to treat rMSCs cultured in vitro. The dynamic assay of alkaline phosphatase (ALP) activity and the number of mineralized nodules labeled with tetracycline were used as the analysis of each group of inducing solution. Whether or not rMSCs can be induced to differentiate into osteoblasts and to promote bone differentiation. Results: On the 12th day of rMSCs culture, the ALP activity of Jianpi Bushen group was higher than that of the blank control group (P<0.05). After the 8th day, the ALP activity of Jianpi Bushen Jiageng was more than that of the osteogenic group (P<0.05). The number of calcium nodules formed in the remaining 3 groups was significantly higher than that in the blank control group after 22d replacement of the corresponding culture fluid (P<0.01); the number of calcium nodules was greater in the spleen tonifying kidney and bone formation group than in the osteogenic group (P<0.05). ). Conclusion: Jianpi Bushen Recipe-containing serum can promote osteogenic differentiation of rMSCs, and has a synergistic effect with classic osteogenic inducers. This may reveal the cytological mechanism of clinical application of this side to prevent steroid-induced femoral head necrosis (SINFH).