中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (6): 872-877.doi: 10.12307/2023.219

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

葛根素与淫羊藿苷对小鼠前成骨细胞生物学性能影响的比较

乔鲁卉1，马子雨2，郭浩宇2，侯玉东2   

  1. 1滨州医学院附属烟台口腔医院修复科，山东省烟台市   264000；2滨州医学院口腔医学院，山东省烟台市   264003
  • 收稿日期:2022-01-04 接受日期:2022-02-22 出版日期:2023-02-28 发布日期:2022-08-11
  • 通讯作者: 侯玉东，硕士，教授，滨州医学院口腔医学院，山东省烟台市 264003
  • 作者简介:乔鲁卉，女，1994年生，汉族，2021年滨州医学院毕业，硕士，医师，主要从事口腔种植修复方面的研究。

Comparison of puerarin and icariin on the biological properties of mouse preosteoblasts

Qiao Luhui1, Ma Ziyu2, Guo Haoyu2, Hou Yudong2   

  1. 1Department of Prosthodontics, Yantai Stomatological Hospital of Binzhou Medical University, Yantai 264000, Shandong Province, China; 2School of Stomatology, Binzhou Medical University, Yantai 264003, Shandong Province, China
  • Received:2022-01-04 Accepted:2022-02-22 Online:2023-02-28 Published:2022-08-11
  • Contact: Hou Yudong, Master, Professor, School of Stomatology, Binzhou Medical University, Yantai 264003, Shandong Province, China
  • About author:Qiao Luhui, Master, Physician, Department of Prosthodontics, Yantai Stomatological Hospital of Binzhou Medical University, Yantai 264000, Shandong Province, China

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摘要：

文题释义：
淫羊藿苷：是中药淫羊藿的有效药理成分之一，属黄酮类物质，可以通过刺激骨形成同时抑制骨吸收来促进骨骼健康。在近年来对淫羊藿苷成骨机制的探索中发现淫羊藿苷能明显提高小鼠前成骨细胞中骨保护蛋白、Runt相关转录因子2基因的表达量，从而促进前成骨细胞分化、成熟。
葛根素：是主要的异黄酮类植物衍生化合物，从葛根中提取，在临床上主要用于医治心脑血管疾病、糖尿病、骨质疏松等疾病，具有类雌激素作用。研究发现，葛根素及其衍生物可作为天然的成骨剂，动物实验研究发现其能防止卵巢切除大鼠的骨质流失等。

背景：在种植区域骨缺损的修复方式中，骨组织工程修复策略相较于传统的自体骨移植方式具有创伤小、无免疫原性以及促进骨再生的优势，将不良反应较化学合成药物小的中药成分作用于骨质疏松患者骨缺损区域的方案是骨组织工程药物递送的重要组成部分。
目的：比较葛根素与淫羊藿苷对小鼠前成骨细胞(MC3T3-E1)增殖、分化和矿化的影响，选择更为有效的促成骨药物。
方法：确定葛根素、淫羊藿苷促MC3T3-E1细胞增殖、分化的最适浓度后将实验细胞分为4组：对照组、17β-雌二醇组和最适浓度葛根素组及淫羊藿苷组。在药物干预后第1，4，7天进行细胞增殖活力检测；成骨诱导第1，7，14天进行碱性磷酸酶活性检测；成骨诱导第21天进行钙化结节茜素红染色；成骨诱导第7天Real Time-PCR检测骨保护蛋白、Runt相关转录因子2 mRNA 的表达水平；鬼笔环肽染色观察培养
24 h的细胞骨架形态。
结果与结论：葛根素与淫羊藿苷促进细胞增殖、分化的最适浓度分别为10-7 mol/L，10-6 mol/L；经比较发现，培养第4天时，与淫羊藿苷组相比，葛根素促进细胞增殖的能力更强，差异有显著性意义(P < 0.05)；成骨诱导7 d时，与葛根素组对比，淫羊藿苷组碱性磷酸酶活性更强，骨保护蛋白、Runt相关转录因子2 mRNA的表达水平上调显著，且在诱导21 d时钙结节形成数量和面积显著增加，差异有显著性意义(P < 0.05)。培养24 h细胞骨架形态呈网格状铺展，与对照组相比，雌激素组、葛根素组、淫羊藿苷组肌动蛋白走行清晰。结果表明，葛根素对MC3T3-E1细胞增殖的促进作用更强，淫羊藿苷对MC3T3-E1细胞分化、矿化促进作用更强。

https://orcid.org/0000-0002-5844-0645(乔鲁卉)

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 葛根素, 淫羊藿苷, 细胞增殖, 细胞分化, 细胞矿化, 成骨细胞

Abstract: BACKGROUND: Compared with the traditional autogenous bone graft methods, the bone tissue engineering repair strategies have their advantages of less trauma, free immunogenicity, and promotion of bone regeneration among the bone defect repair modes in the implant area. The scheme that applies the traditional Chinese medicine ingredients with less adverse reactions than chemical synthetic drugs to the bone defect area of osteoporosis patients constitutes an important part of the bone tissue engineering scaffold to deliver drugs.
OBJECTIVE: To compare effects of puerarin and icariin on the proliferation, differentiation and mineralization of mouse preosteoblasts (MC3T3-E1) and select effective osteogenic drugs.
METHODS:  The optimal concentration of puerarin and icariin to promote the proliferation and differentiation of MC3T3-E1 was determined. Cultured cells were divided into control group, 17β-estradiol group, icariin group, and puerarin group. Cell proliferation and viability were detected at 1, 4, and 7 days after drug intervention. Alkaline phosphatase activity was detected at 1, 7, and 14 days after osteogenic induction. Calcified nodules after alizarin red staining were measured at 21 days after osteogenic induction. Real time-qPCR was used to detect the expression of osteoprotegerin and Runt-associated transcription factor 2 mRNA at 7 days after osteogenic induction. The morphology of the cytoskeleton after 24 hours was observed using phalloidin staining.
RESULTS AND CONCLUSION: Puerarin and icariin exhibited the best effect on proliferation and osteogenic differentiation at concentration of 10-7 mol/L and 10-6 mol/L. At 4 days of culture, compared with the icariin group, the ability of puerarin to promote cell proliferation was stronger, and the difference was significant (P < 0.05). At 7 days of osteogenic induction, compared with the puerarin group, the alkaline phosphatase activity of the icariin group was stronger; the expression levels of osteoprotegerin and Runt-related transcription factor 2 mRNA were significantly up-regulated; at 21 days of induction, the number and area of calcified nodules increased significantly, and the difference was significant (P < 0.05). After 24 hours of culture, the cytoskeletal morphology was grid-like spreading. Compared with the control group, the actin ran clearly in the estrogen group, the puerarin group and the icariin group. According to the results, puerarin can better accelerate the proliferation of MC3T3-E1 cells, and icariin can better promote the differentiation and mineralization of MC3T3-E1 cells.

Key words: puerarin, icariin, cell proliferation, cell differentiation, cell mineralization, osteoblast

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