Page 45 - 《中国药房》2023年7期

木犀草素对骨缺损成骨修复的作用及机制研究
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汤胜尧 ，胡珉华 ，周若愚 ，孙伟鹏 ，唐鑫陶 ，林海雄 ，姜自伟 （1.广州中医药大学第一临床医学院，广州
510405；2.宁夏回族自治区中医医院暨中医研究院，银川 750021；3.香港中文大学组织工程与再生医学研究
所，香港 999077；4.广州中医药大学第一附属医院一骨科，广州 510405）
中图分类号 R965；R285 文献标志码 A 文章编号 1001-0408（2023）07-0807-07
DOI 10.6039/j.issn.1001-0408.2023.07.08
摘 要 目的 探讨木犀草素对骨缺损成骨修复的作用及机制。方法 通过网络药理学方法筛选木犀草素治疗骨缺损的作用靶点
及潜在通路，利用 Hub 基因筛选排名前 2 位的靶点进行分子对接验证，以结合能作为评判标准。对大鼠骨髓间充质干细胞
（BMSC）及脐静脉内皮细胞（RUVEC）进行体外实验，采用碱性磷酸酶染色、茜素红S染色、体外血管形成实验进行表型验证，采用
Western blot法检测磷脂酰肌醇3激酶（PI3K）、蛋白激酶B1（Akt1）蛋白的表达情况，以验证木犀草素对BMSC成骨分化及RUVEC
体外血管形成的作用机制。结果 网络药理学结果显示，木犀草素对骨缺损的血管形成和骨修复作用主要与Akt1、SRC蛋白、雌
激素受体1、表皮生长因子受体、环加氧酶2、基质金属蛋白酶9靶点有关，且与PI3K/Akt信号通路关系密切。分子对接结果显示，
木犀草素与Akt1、SRC蛋白结合稳定。体外实验结果表明，木犀草素能显著增高BMSC内碱性磷酸酶的表达水平和活性，增加钙
盐沉积和钙化结节的数量，促进BMSC的钙化；与木犀草素0 μmol/L组比较，木犀草素1、10 μmol/L组的RUVEC血管形成能力显著
增强、血管长度显著增加，PI3K、Akt1的蛋白表达水平显著提高（P＜0.05或P＜0.01），且浓度越高效果越好。结论 木犀草素可能
通过激活PI3K/Akt信号通路，促进PI3K、Akt1蛋白的表达来发挥增强骨折端的血管形成及骨修复的功能。
关键词 木犀草素；骨缺损；成骨修复；血管形成；网络药理学；分子对接

Study on the effects and mechanism of luteolin on osteogenic repair of bone defects
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TANG Shengyao ，HU Minhua ，ZHOU Ruoyu ，SUN Weipeng ，TANG Xintao ，LIN Haixiong ，JIANG Ziwei 4
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（1. The First Clinical Medical College， Guangzhou University of Chinese Medicine， Guangzhou 510405， China；
2. Ningxia Hui Autonomous Region Hospital of Traditional Chinese Medicine&Academy of Traditional Chinese
Medicine， Yinchuan 750021， China；3. Institute of Tissue Engineering and Regenerative Medicine， the Chinese
University of Hong Kong， Hong Kong 999077， China；4. Dept. One of Orthopedics， the First Affiliated Hospital
of Guangzhou University of Chinese Medicine， Guangzhou 510405， China）
ABSTRACT OBJECTIVE To investigate the effects and mechanism of luteolin on osteogenic repair of bone defects.
METHODS The targets and potential pathways of luteolin in the treatment of bone defects were screened by network pharmacology
method， and then the top 2 targets were selected by Hub gene screening for molecular docking verification， with binding energy as
the evaluation standard. In vitro experiments were conducted on rat bone mesenchymal stem cells （BMSC） and rat umbilical vein
endothelial cells （RUVEC）. Phenotypic validation was performed using alkaline phosphatase staining， alizarin red S staining， and
in vitro angiogenesis experiments. Western blot assay was used to detect the protein expressions of phosphatidylinositol 3 kinase
（PI3K） and protein kinase 1 （Akt1）， so as to validate the mechanism of luteolin on osteogenic differentiation of BMSC and
angiogenesis of RUVEC in vitro. RESULTS The results of network pharmacology showed that the effects of luteolin on vascular
formation and bone repair in bone defects were mainly related to Akt1， SRC， estrogen receptor 1， epidermal growth factor
receptor， cyclooxygenase 2， matrix metalloproteinase 9 targets， and were closely related to PI3K-Akt signaling pathway. The
results of molecular docking showed that luteolin binding to Akt1 and SRC proteins was stable. The results of in vitro experiments
showed that luteolin could significantly improve the expressions and activities of alkaline phosphatase in BMSC， increased the
number of calcium salt deposits and calcified nodules， and promoted calcification of BMSC. Compared with luteolin 0 μmol/L
group， the angiogenesis ability of RUVEC was enhanced
Δ 基金项目 国家自然科学基金资助项目（No.8197150751）；宁夏 significantly in luteolin 1， 10 μmol/L groups， the length of
自然科学基金项目（No.2022AAC03398） blood vessels and the protein expressions of PI3K and Akt1
＊第一作者 硕士研究生。研究方向：中医药防治骨与关节损伤。 were significantly increased （P＜0.05 or P＜0.01）； the higher
E-mail：1315492593@qq.com
# 通信作者 主任中医师，教授，博士。研究方向：中医药防治骨与 the concentration， the better the effect. CONCLUSIONS
关节损伤。E-mail：ainemylyy@163.com Luteolin may play a role in promoting angiogenesis and bone

中国药房 2023年第34卷第7期 China Pharmacy 2023 Vol. 34 No. 7 · 807 ·