淫羊藿黄酮类主要成分促进骨髓间充质干细胞向成骨细胞增殖分化作用及机制的影响

许静; 张晶晶; 郭非非; 李欣; 张毅; 杨洪军; 吴宏伟; 张琼

doi:10.13422/j.cnki.syfjx.2017140113

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淫羊藿黄酮类主要成分促进骨髓间充质干细胞向成骨细胞增殖分化作用及机制的影响

药理 | 更新时间：2024-01-04

- 淫羊藿黄酮类主要成分促进骨髓间充质干细胞向成骨细胞增殖分化作用及机制的影响
- Promotive Effect of Epimedii Folium Flavonoids on Proliferation and Differentiation of BMSCs into Osteoblasts
- 中国实验方剂学杂志 2017年23卷第14期页码：113-120
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家科技支撑计划项目（2011BAI07B08）
- DOI：10.13422/j.cnki.syfjx.2017140113
  中图分类号： R285
- 纸质出版日期：2017
- 稿件说明：
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许静, 张晶晶, 郭非非, 等. 淫羊藿黄酮类主要成分促进骨髓间充质干细胞向成骨细胞增殖分化作用及机制的影响[J]. 中国实验方剂学杂志, 2017,23(14):113-120.

XU Jing, ZHANG Jing-jing, GUO Fei-fei, et al. Promotive Effect of Epimedii Folium Flavonoids on Proliferation and Differentiation of BMSCs into Osteoblasts[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(14): 113-120.
许静, 张晶晶, 郭非非, 等. 淫羊藿黄酮类主要成分促进骨髓间充质干细胞向成骨细胞增殖分化作用及机制的影响[J]. 中国实验方剂学杂志, 2017,23(14):113-120. DOI： 10.13422/j.cnki.syfjx.2017140113.

XU Jing, ZHANG Jing-jing, GUO Fei-fei, et al. Promotive Effect of Epimedii Folium Flavonoids on Proliferation and Differentiation of BMSCs into Osteoblasts[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(14): 113-120. DOI： 10.13422/j.cnki.syfjx.2017140113.

摘要

目的：探讨淫羊藿黄酮类主要成分朝藿定A，朝藿定B，朝藿定C，淫羊藿苷，淫羊藿次苷对骨髓间充质干细胞（BMSCs）向成骨细胞分化的促进作用，并对其作用机制进行预测。方法：采用全骨髓细胞贴壁法分离纯化SD大鼠BMSCs做原代培养。实验分为朝藿定A+成骨诱导培养基（1×10-8 mol·L-1）组，朝藿定B+成骨诱导培养基（1×10-8 mol·L-1）组，朝藿定C+成骨诱导培养基（1×10-8 mol·L-1）组，淫羊藿苷+成骨诱导培养基（1×10-8 mol·L-1）组，淫羊藿次苷组+成骨诱导培养基组及成骨诱导培养基组（空白组）6组。加入药物进行干预14 d后，采用定性与定量相结合的方法，通过茜素红染色、细胞骨架染色、碱性磷酸酶定量分析，对淫羊藿5种黄酮类主要成分促进骨髓间充质干细胞向成骨细胞分化作用进行药效学评价；在此基础上，基于BATMAN-TCM网络药理学研究平台，对相关靶点和通路进行预测分析，探讨其潜在作用机制。结果：BMSCs在作用浓度为1×10-8 mol·L-1的药物浓度作用下诱导14 d，通过细胞骨架染色，各给药组均可明显观察到多角形成骨细胞增多；通过茜素红染色，各给药组可明显观察到大量的矿化结节的形成；通过碱性磷酸酶含量测定，与空白组比较，各给药组碱性磷酸酶含量升高，其中朝藿定C，淫羊藿苷组有显著性差异（P<0.05），淫羊藿次苷组具有极显著性差异（P<0.01）。同时网络药理学分析表明，5种黄酮类成分在促进增殖和分化方面共得到33个潜在作用靶点，并富集分析得到8条信号通路，其中activation of MAPK activity，estrogen signaling pathway已有实验报道。结论：淫羊藿黄酮类主要成分朝藿定A，朝藿定B，朝藿定C，淫羊藿苷，淫羊藿次苷均具有促进BMSCs向成骨方向分化的作用，其中朝藿定C，淫羊藿苷，淫羊藿次苷的作用效果显著，网络药理学分析结果从整体上阐释了多成分的作用机制，为进一步实验验证提供参考。

Abstract

Objective: To investigate the effects of main Epimedii Folium flavonoids (epimedin A

epimedin B

epimedin C

icariin and icarisid Ⅰ) on promoting BMSCs proliferation and differentiation into osteoblasts and to predict the mechanism of them. Method: BMSCs in SD rats were isolated and purified by whole bone marrow adherence method for primary culture. The experiment was divided into 6 groups: epimedin A+ osteogenic medium (1×10-8 mol·L-1)

epimedin B+ osteogenic medium (1×10-8 mol·L-1)

epimedin C+ osteogenic medium (1×10-8 mol·L-1)

icariin + osteogenic medium (1×10-8 mol·L-1)

icariside group + osteogenic medium (blank group). After drug intervention for 14 days

qualitative and quantitative methods were combined for pharmacodynamic evaluation

of the above 5 kinds of Epimedii Folium flavonoids by alizarin red staining

cytoskeleton staining and alkaline phosphatase assay. Meanwhile

the potential targets and pathways were predicted and analyzed based on BATMAN-TCM network pharmacology research platform to investigate the action mechanisms. Result: BMSCs were treated with 1×10-8 mol·L-1 drugs for 14 d

and polygonal osteoblasts were increased in various groups by cytoskeleton staining; in alizarin red staining

a large number of mineralized nodules were formed in various treatment groups; in Alkaline phosphatase assay

the content of alkaline phosphatase was increased in various treatment groups as compared with the blank group

particularly

the groups of epimedin C(P<0.05)

icariin (P<0.05)

and icarisid Ⅰ(P<0.01) showed significant changes. As for network pharmacology analysis

33 potential targets and 8 pathways about proliferation and differentiation were obtained. Among them

activation of MAPK activity and estrogen signaling pathway had been verified by the relative experimental reports. Conclusion: All of the above 5 kinds of Epimedii Folium flavonoids could promote BMSCs proliferation and differentiation into osteoblasts. especially

epimedin C

icariin

and icarisid Ⅰ had significant effects. The results of network pharmacology analysis interpreted the overall mechanism of the Epimedii Folium flavonoids

providing a reference for further research.

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