骨碎补提取物对骨髓间充质干细胞向成骨细胞分化及Cbfα1表达的影响

摘要

Objective To investigate the effect of drynaria extract on the proliferation and differentiation of rat bone marrow mesenchymal stem cells (BMSCs) and expression of Cbfα1.Methods With the method of interference of Chinese medicine cell culture,single drynaria combined with medium was used to coculture the rat BMSCs.The rat BMSCs were divided into 5 groups:the standard medium group (serum-free culture medium),conditioned medium group(dexamethasone 1 × 10-8 mol/L,β-glycerophosphate salt 10 mol/L,VitC 50 μg/mL),traditional Chinese medicine group 1 (drynaria extract 20 mg/mL),TCM group 2(drynaria extract 2 mg/mL) and TCM group 3 (drynaria extrat 0.2 mg/mL) ; the potential of osteogenic differentiation of BMSCs was detected by type Ⅰ collagen immumohistochemical staining; and the expression of Cbfα1 was detected by RT-PCR for semi-quantitative analysis.Results Type Ⅰ collagen immumohistochemical staining and RT-PCR showed that the potential of osteogenic differentiation of BMSCs and the expression of Cbfα1 in the drynaria group were much better than those in the blank group and conditioned group (P ＜ 0.01),and the effect of 2 mg/mL drynaria group was superior to those in 20 mg/mL and 0.2 mg/mL drynaria group (P ＜ 0.05).Conclusion Traditional Chinese medicine drynaria could promote the proliferation and osteogenic differentiation of BMSCs and increase the expression of Cbfα1.%目的 探讨中药骨碎补对大鼠骨髓间充质干细胞增殖分化及核心结合因子α1(Cbfα1)表达的影响.方法 采用中药干预培养细胞的方法,利用骨碎补单味药物与培养基联合培养大鼠骨髓间充质干细胞(BMSCs),分为标准培养基组(无血清培养基)、条件培养基组(地塞米松1×10-8 mol/L、β-甘油磷酸钠10 mol/L、VitC 50 μg/mL)、中药1组(骨碎补提取液20 mg/mL)、中药2组(骨碎补提取液2 mg/mL)、中药3组(骨碎补提取液0.2 mg/mL)共5组,采用Ⅰ型胶原免疫组化染色检测骨碎补促BMSCs成骨分化能力,RT-PCR方法进行半定量分析Cbfα1表达.结果 中药1、2、3组BMSCs成骨分化能力及Cbfα1表达均明显优于空白组及条件培养组(P＜0.01),2 mg/mL药物组优于20 mg/mL及0.2 mg/mL药物组(P＜0.05).结论 体外中药骨碎补可促进大鼠BMSCs增殖及成骨分化,加强Cbfα1表达.