摘要
目的:通过观察骨碎补水提液对SD大鼠骨髓间充质干细胞(MSCs)骨向分化能力及转化生长因子β1(TGF-β1)、骨形态发生蛋白2(BMP-2)表达的影响,阐释其可能的诱导机制。方法:运用全骨髓贴壁法分离、纯化大鼠MSCs;运用MTT法检测不同浓度的骨碎补水提液对SD大鼠MSCs增殖活力的影响;以ALP活性及ALP染色阳性率确定骨碎补水提液最佳促MSCs骨向分化浓度,并以该浓度对MSCs骨向分化进行干预,按是否添加经典成骨诱导液将实验分为:空白对照组、经典成骨诱导液组、骨碎补水提液组、骨碎补水提液+经典成骨诱导液组,通过检测碱性磷酸酶(ALP)、Ⅰ型胶原(ColⅠ)、骨钙素(BGP)和钙化结节的表达,评价各组骨向分化能力;通过检测转化生长因子β1(TGF-β1)、骨形态发生蛋白2(BMP-2)的表达,探讨药物促MSCs骨向分化的作用机制。结果:骨碎补水提液最佳促MSCs增殖浓度为5μg/mL;最佳促MSCs骨向分化浓度为50μg/mL;经典成骨诱导液、骨碎补水提液及联合用药均能促进MSCs骨向分化及TGF-β1和BMP-2的表达。结论:补肾中药骨碎补能有效的促进MSCs增殖和骨向分化。在MSCs骨向分化过程中,上调TGF-β1、BMP-2的表达量可能是诱导各组促进MSCs骨向分化的作用机制之一。
Objective:To investigate the effects of Drynariae Rhizoma water-extraction on the ability of osteogenic differentiation and the expression of TGF-β1 and BMP-2 in inducing mesenchymal stem cells (MSCs) differentiation into osteoblast and its mechanism. Methods:MSCs were isolated and purified by differential time adherent method ;The most effective concentration of the Drynariae Rhizoma water-extraction on the proliferation and ostogenic differentiation of MSCs was confirmed by MTT method and the activity and positive staining of alkaline phosphatase (ALP)respectively;According to different induced condition, MSCs were divided into 4 groups: control group, classic group( induced by the classic osteoblast-induced system), Drynariae Rhizoma water-extraction group (induced by the most effective concentration of Drynariae Rhizoma water-extraction on differentiation into osteoblast) and Drynariae Rhizoma water- extraction + classic group( induced by the combination of classic osteoblast-induced system and the most effective concentration of Dry- nariae Rhizoma water-extraction on differentiation into osteoblast). ALP, type I collagen (Col I ), bone gla protein (BGP) and calcium nodes in each group were detected and compared to indicate the osteogenic differentiation of each group. TGF-β1, and BMP-2 were detected by ELISA. Results:The most effective concentration of the Drynariae Rhizoma water-extraction on the proliferation and osteogenic differentiation of MSCs was 5 μg/mL and 50 μg/mL, respectively. The classic group, Drynariae Rhizoma water-extraction group and Drynariae Rhizoma water-extraction + classic group could promote the osteogenic differentiation of MSCs and increase the expression of TGF-β1 and BMP-2. Conclusion:The Drynariae Rhizoma water-extraction can promote the proliferation and osteogenic differentiation of MSCs. Epimedium water-extraction can improve the differentiation of MSC into osteoblast and its mechanism may be related to increasing the expression of TGF-β1 and BMP-2
出处
《中药材》
CAS
CSCD
北大核心
2013年第8期1287-1292,共6页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金项目(81173619)
广东省教育厅育苗工程(LYM10025)