摘要
目的探讨骨碎补总黄酮对骨髓间充质干细胞（BMSC）成骨分化过程中，对Wnt／β-catenin信号通路相关因子mRNA表达的影响。方法采用全骨髓贴壁法分离、培养sD大鼠BMSC，取3代细胞予100tLg／ml浓度骨碎补总黄酮进行干预，21d后，观察并行矿化结节染色，在干预7、14、21、28d时检测成骨分化标志物碱性磷酸酶（ALP）活性和Wnt／β-catenin信号通路相关因子β-catenin、淋巴增强因子1（LEF-1）、细胞周期蛋白D（cyclineD）mRNA的表达情况。结果干预后各个时间点骨碎补总黄酮组ALP活性（U／L）均较空白组高（7d：11．10±0．08比1．61±0．14；14d：24．62±0．34比1．64±0．04：21d：18．4±0．06比1．53±0．04；28d：14．91±0．14比1．52±0．04；均P〈0．01）；21d后行矿化结节染色，骨碎补总黄酮组为阳性，空白组为阴性；干预后β—cateninmRNA在第14天表达最高（0．357±0．062比0．174±0．013，P〈0．05）；LEF-1、cyclineDmRNA在第7天表达最高（LEF-1：0．0611±0．0002比0．0345±0．0131；cyclineD：0．1510±0．0255比0．0718±0．0294均P〈0．05）。结论骨碎补总黄酮诱导大鼠BMSC向成骨细胞分化，伴随Wnt／β-catenin信号通路相关因子mRNA表达的变化。
Objective To explore the effects of the expression of Wnt/β-catenin signaling factor mRNA during drynaria total flavonoids on the osteogenic differentiation of bone marrow mesenchymal stem cells （BMSCs）. Methods The BMSCs were isolated from SD rats by whole bone marrow culture method and purified by passage. And the P3 BMSCs were intervened with 100 μg/ml drynaria total flavonoids. At Day 21, mineralized staining was performed. At Days 7, 14, 21 and 28 post-intervention, the activity of alkaline phosphatase （ALP） was detected and polymerase chain reaction （PCR） used to detect the expressions of Wnt/β-catenin signaling pathway related factors β-catenin, LEF-1 and cycline D mRNA. Results At each timepoint post-intervention, comparing the ALP activity in cell supernatant between two group, the drynaria total flavonoids group was higher than the blank control group （7 d： 11.10 ± 0. 08 vs 1.61±0.14; 14 d ： 24. 62± 0. 34 vs1. 64 ± 0. 04 ; 21d ： 18. 41± 0. 06 vs l. 53±0.04; 28 d： 14.9±0. 14 vs 1.52±0. 04 ; all P 〈 0. 01 ）. At Day 21, upon staining with alizarin red, the drynaria total flavonoids group was positive while the blank control group negative. At Day 14, the expression of β-catenin mRNA was higher in the drynaria total flavonoids group higher than that in the blank control group （0. 357 ±0. 063 vs 0. 174±0. 013, P 〈0. 05）. At Day 7, the expressions of LEF-1 and cyeline D mRNA were higher in the drynaria total flavonoids group than those in the blank control group （LEF-1 0. 0611 ±0. 0002 vs 0. 0345 ±0. 0131 ; cycline D 0. 1510 ±0. 0255 vs 0. 0718 ± 0. 0294, all P 〈 0. 05 ）. Conclusion Drynaria total flavonoids induce BMSCs to differentiate into osteoblasts. And it is accompanied with the altered expression of Wnt/β-catenin signaling pathway related factor mRNA.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2012年第32期2288-2291,共4页
National Medical Journal of China
基金
国家中医药管理局科研基金（04_05zP51）
广东省中医药管理局科研基金（2007263）
广东珠海市医学科研基金（2011044）
关键词
骨质疏松
骨碎补总黄酮
骨髓间充质干细胞
成骨分化
Osteoporosis
Drynaria total flavonoids
Bone marrow mesenchymal stem cells
Osteogenic differentiation
