赖满香　冯娟　阮志燕

[摘要] 目的 探讨巴戟天多糖含药血清对SD大鼠骨髓间充质干细胞（BMSCs）骨向分化过程中核心结合因子ɑ-1（Cbfɑ-1）mRNA表达的影响。 方法 全骨髓贴壁法分离培养SD大鼠BMSCs，采用pNPP法检测不同浓度r巴戟天多糖含药血清（高、中、低）对BMSCs碱性磷酸酶（ALP）活性的影响，并以其最佳作用浓度进行后续实验，后续实验分成空白组、成骨诱导组、巴戟天多糖含药血清组、联合组（巴戟天多糖含药血清组+成骨诱导组），各组用药14d，采用RT-PCR技术检测各组BMSCs Cbfa-1 mRNA表达的情况。 结果 与对照组比较，不同剂量的巴戟天多糖含药血清均可提高BMSCs分泌ALP（F=126.278，P<0.05），其中高剂量组的作用强度高于其它剂量组；与空白组相比，成骨诱导组、巴戟天多糖含药血清组、联合组均能上调BMSCs中Cbfa-1 mRNA的表达（F=261.412，P<0.05）；但与成骨诱导组相比，巴戟天多糖含药血清组不及成骨诱导组，两者差异有统计学意义（t=26.721，P<0.05）。 结论 巴戟天多糖含药血清能够促进BMSCs向成骨细胞分化，并能上调Cbfa-1 mRNA的表达，但其作用不及成骨诱导强。

[关键词] 巴戟天多糖含药血清；骨髓间充质干细胞；碱性磷酸酶；骨向分化；Cbfa-1

[中图分类号] R285.5 [文献标识码] A [文章编号] 2095-0616（2017）15-27-04

[Abstract] Objective To investigate the effects of morindae officinalis polysaccharide （MOP） containing serum on Cbfa-1 mRNA expression during the differentiation from BMSCs to the osteoblast. Methods BMSCs were isolated by whole bone marrow adherent method.Different concentrations of MOP-containing serum，including high dosage，middle dosage and low dosage，were added to BMSCs section to detect the alkaline phosphatase （ALP） activity by pNPP method，the best dosage was added to the next experiment.BMSCs were divided into the blank group，the osteogenic induced group，the MOP-containing serum group，MOP-containing serum and the osteogenic induced co-culture group.Cbfa-1 mRNA in each group were detected by RT-PCR at day 14. Results Compared with the control group，different concentrations of MOP-containing serum could elevate serum ALP activities（F=126.278，P<0.05），and the high dosage was the best.Compared with the blank group，Cbfa-1 mRNA expression levels increased in the osteogenic induced group，MOP-containing serum group and co-culture group， difference has statistical significance（F=261.412， P<0.05）.Cbfa-1 mRNA expression levels was lower than the osteogenic induced group， difference had statistical significance（t=26.721，P<0.05）. Conclusion The MOP-containing serum can promote the differentiation of BMSCs to osteoblasts，and increase Cbfa-1 mRNA expression，but its strength is lower than the osteogenic induced.

[Key words] MOP-containing serum；Bone marrow mesenchymal stem cell；Alkaline phosphatase；Osteogenic differentiation；Cbfa-1

巴戟天為茜草科多年生藤植物，为我国著名的四大南之一，现代药理研究证明：巴戟天具有强壮骨骼、调节免疫功能、调节甲状腺功能、抗衰老、抗

疲劳、增强记忆、抗肿瘤及促进造血等作用[1]。巴戟天多糖（morindae officinalis polysaccharide，MOP）为巴戟天的有效成分之一，有实验研究证实，巴戟天多糖具有预防骨质疏松的作用[2-3]，体外能促进成骨细胞增殖，提高成骨细胞分泌ALP与骨钙素，抑制破骨细胞活性[4]，巴戟天提取物能促进MSCs向成骨细胞分化，并能加强Cbfa1的表达[5]，但巴戟天作用于MSCs的具体成分不清。本实验观察巴戟天多糖含药血清对SD大鼠BMSCs向成骨分化过程中核心结合因子ɑ-1（Cbfɑ-1）表达的影响，从干细胞水平探讨巴戟天多糖促BMSCs骨向分化中的可能作用机制。

1 材料与方法

1.1 实验动物

实验所用动物为清洁级3月龄SD大鼠40只，体重245～255g；……