摘要
目的探讨淫羊藿总黄酮对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)成骨分化过程中BMP-2/RunX2/OSX信号通路的影响。方法将培养的第3代BMSCs随机分为对照组、实验组及抑制剂组,对照组细胞予0.2%二甲基亚砜加OS液加DMEM/F12培养基培养,实验组加入20μg/mL淫羊藿总黄酮进行干预,抑制剂组予20μg/mL淫羊藿总黄酮加1μg/mL NOGGIN重组蛋白干预,9天后测定碱性磷酸酶(ALP)活性,茜素红染色法染色并观察钙结节密度,RT-PCR法检测成骨相关蛋白(Ⅰ型胶原、骨钙蛋白及骨桥蛋白)及BMP-2/RunX2/OSX信号通路相关因子的转录表达。结果与对照组比较,实验组经定向成骨诱导后BMSCs ALP活性增强,钙结节密度显著增大,Ⅰ型胶原、骨钙蛋白和骨桥蛋白表达量增加,成骨相关转录因子BMP-2、RunX2和OSX的表达量亦增加(P<0.05)。与实验组比较,抑制剂组BMSCs ALP活性降低(P<0.05),钙结节密度降低,Ⅰ型胶原、骨钙蛋白及骨桥蛋白表达水平降低(P<0.05)。结论 20μg/mL淫羊藿总黄酮能通过BMP-2/RunX2/OSX信号通路促进BMSCs定向成骨分化的进程。
Objective To explore the effect of total flavonoids of Herba Epimedium（FHE） on BMP-2/Run X2/OSX signaling pathway in promoting osteogenic differentiation of bone marrow mesenchymal stem cells（BMSCs）. Methods Passage 3 BMSCs were randomly divided into the control group, the experimental group, and the inhibitor group. BMSCs in the control group were cultured in 0. 2% dimethyl sulfoxide ＋ Osteogenuxic Supplement（OS） fluid ＋ DMEM/F12 culture media. BMSCs in the experimental group were intervened by 20 μg/m L FHE.BMSCs in the inhibitor group were intervened by 20 μg/m L FHE and 1 μg/m L NOGGIN recombinant protein. At day 9 alkaline phosphatase（ALP） activity was measured. Calcium nodules were stained by alizarin red staining and the density was observed. The transcription expression of osteogenic differentiation-related proteins（type Ⅰcollagen, osteocalcin, and osteopontin） and related factors of BMP-2/Run X2/OSX signaling pathway was assayed by RT-PCR. Results Compared with the control group, ALP activities were enhanced and the density of calcium nodules significantly increased;type Ⅰ collagen, osteocalcin, and osteopontin expression levels were increased in the experimental group. The expression of osteogenesis-related transcription factor was also increased in the experimental group. Noggin recombinant protein inhibited FHE promoting BMSCs osteogenesis in the inhibitor group. Compared with the experimental group, ALP activity decreased（P〈0. 05）, the density of calcium noduleswas lowered, expression levels of type Ⅰ collagen, osteocalcin, osteopontin significantly decreased（P〈0. 05） in the inhibitor group. Conclusion 20 μg/m L FHE promoted osteogenic differentiation process of BMSCs by BMP-2/Run X2/OSX signaling pathway.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2016年第5期614-618,共5页
Chinese Journal of Integrated Traditional and Western Medicine
