淫羊藿次苷对犬颌骨间充质干细胞增殖与成骨分化的影响

doi:10.19438/j.cjoms.2019.04.004

中国口腔颌面外科杂志 ›› 2019, Vol. 17 ›› Issue (4): 304-310.doi: 10.19438/j.cjoms.2019.04.004

淫羊藿次苷对犬颌骨间充质干细胞增殖与成骨分化的影响

张意闻¹, 罗远^1,2, 郭华艳³, 张迎娣³, 缪国俊³, 黄远亮³

1.同济大学口腔医学院口腔颌面外科教研室,上海牙组织与再建工程技术研究中心,上海 200072;
2.上海市口腔病防治院口腔颌面外科,上海 200031;
3.同济大学附属东方医院口腔科,上海 200120

收稿日期:2018-12-03 修回日期:2019-01-21 出版日期:2019-07-20 发布日期:2019-08-12
通讯作者: 黄远亮,E-mail: ylhuang0115@163.com
作者简介:张意闻（1993-）,女,硕士研究生,E-mail: zhangyiwen0129@163.com
基金资助:
国家自然科学基金（81771037）; 上海市浦东新区卫生系统特色专病学科建设基金（PWZzb2017-16）

Effects of Icarisid Ⅱ on proliferation and osteogenic differentiation of alveolar bone-derived stem cells in Beagle dogs

ZHANG Yi-wen¹, LUO Yuan^1,2, GUO Hua-yan³, MIAO Guo-jun³, ZHANG Ying-di³, HUANG Yuan-liang³

1.Department of Oral and Maxillofacial Surgery, School of Stomatology, Tongji University; Shanghai Engineering Research Center of Tooth Restoration and Regeneration. Shanghai 200072;
2.Department of Oral Surgery, Shanghai Stomatology Disease Centre. Shanghai 200031;
3.Department of Stomatology, Oriental Hospital, Tongji University. Shanghai 200120,China

Received:2018-12-03 Revised:2019-01-21 Online:2019-07-20 Published:2019-08-12

摘要/Abstract

摘要： 目的研究淫羊藿次苷Ⅱ（Icarisid Ⅱ,ICSII）对体外培养的比格犬颌骨间充质干细胞（alveolar bone-derived stem cells, AMSCs）增殖与成骨分化的影响。方法取比格犬颌骨骨片,体外分离培养出dAMSCs,传代至第4代,做多向分化鉴定。以10^-5、10^-6、10^-7、10^-8、10^-9 mol/L浓度的ICSII刺激dAMSCs后,取1、3、5、7 d的细胞,分别用CCK-8及碱性磷酸酶（ALP）试剂盒检测dAMSCs的增殖及ALP活性;用10^-6 mol/L的ICSII处理dAMSCs后14 d,作ALP染色,21 d时作茜素红染色,以判断ICSII对细胞分化及矿化能力的影响。在成骨诱导及10^-6 mol/L的ICSII药物诱导第4、8天后,用蛋白免疫印迹试验分析成骨相关蛋白OSX、Runx-2及OCN的表达量;在诱导第6天时,利用RT-PCR分析成骨相关基因OSX、bFGF、Runx-2及OCN的表达情况。采用SPSS 22.0软件包进行统计分析。结果原代培养的dAMSCs贴壁生长,呈梭形,具备多向分化能力;ICSII在不同浓度下均可促进dAMSCs的增殖,其ALP活性亦有提高,且在实验浓度时可观察到钙结节形成。各成骨相关基因在ICSII诱导后表达量有不同程度增加,均高于完全培养基组。随着时间延长,加药组Runx-2蛋白表达量有所下降, OCN蛋白表达量逐渐增多,与对照组差距加大;OSX的蛋白表达量虽高于对照组,但无统计学差异。结论 ICSII可促进dAMSCs的增殖及成骨分化。

关键词: 淫羊藿次苷Ⅱ, 颌骨间充质干细胞, 比格犬, 增殖, 分化

Abstract: PURPOSE: To investigate the effect of Icarisid Ⅱon proliferation and osteogenic differentiation of alveolar bone-derived stem cells(AMSCs) in Beagle dogs. Methods: Alveolar bone-derived stem cells were isolated and cultured from the jaw bone of Beagle dogs in vitro. After dAMSCs were stimulated by ICSII at 10^-5, 10^-6, 10^-7, 10^-8 and 10^-9 mol/L, the proliferation and alkaline phosphatase (ACP) activity of dAMSCs were detected by CCK-8 and ALP kit at 1, 3, 5 and 7 days. ALP staining, Alizarin red staining, Western blot and RT-PCR were performed at different time points to evaluate the osteogenic capacity of cells. SPSS22.0 software package was used for statistical analysis. Results: Primary dAMSCs adhered to the wall, showing spindle shape and multidirectional differentiation ability. ICSII can promote the proliferation of dAMSCs at different concentrations. The results showed that ALP activity was also increased, and the formation of calcium nodules was observed at the experimental concentration. The expression levels of each osteogenic gene increased, which were significantly higher than that of the complete medium group. With the extension of time, runx-2 protein expression decreased in the experimental group, OCN protein expression gradually increased, and the gap between the experimental group and the control group widened. OSX protein expression was higher than that of the control group, but there was no significant difference. Conclusions: ICSII can promote the proliferation and osteogenic differentiation of dAMSCs.

Key words: Icarisid Ⅱ, Alveolar bone-derived stem cells, Beagle dogs, Proliferation, Osteogenic differentiation

中图分类号:

R782.2

张意闻, 罗远, 郭华艳, 张迎娣, 缪国俊, 黄远亮. 淫羊藿次苷对犬颌骨间充质干细胞增殖与成骨分化的影响[J]. 中国口腔颌面外科杂志, 2019, 17(4): 304-310.

ZHANG Yi-wen, LUO Yuan, GUO Hua-yan, MIAO Guo-jun, ZHANG Ying-di, HUANG Yuan-liang. Effects of Icarisid Ⅱ on proliferation and osteogenic differentiation of alveolar bone-derived stem cells in Beagle dogs[J]. China J Oral Maxillofac Surg, 2019, 17(4): 304-310.

参考文献

[1] Liu H, Xiong Y, Wang H, et al.Effects of water extract from epimedium on neuropeptide signaling in an ovariectomized osteoporosis rat model[J]. J Ethnopharmacol, 2018, 221:126-136.
[2] Liu M, Xu H, Ma Y, et al.Osteoblasts proliferation and differentiation stimulating activities of the main components of epimedii folium[J]. Pharmacogn Mag, 2017, 13(49): 90-94.
[3] Luo G, Xu B, Huang Y.Icariside II promotes the osteogenic differentiation of canine bone marrow mesenchymal stem cells via the PI3K/AKT/mTOR/S6K1 signaling pathways[J]. Am J Transl Res, 2017, 9(5): 2077-2087.
[4] 迟海洋, 刘晓奇, 闫井龙. 细胞因子与骨髓间充质干细胞在骨组织工程中的研究现状[J]. 医学综述, 2017, 23(24): 4796-4800.
[5] Mason S, Tarle SA, Osibin W, et al.Standardization and safety of alveolar bone-derived stem cell isolation[J]. J Dent Res, 2014, 93(1): 55-61.
[6] Caplan AI.Adult mesenchymal stem cells for tissue engineering versus regenerative medicine[J]. J Cell Physiol, 2007, 213(2): 341-347.
[7] Park JC, Kim JC, Kim YT, et al.Acquisition of human alveolar bone-derived stromal cells using minimally irrigated implant osteotomy: in vitro and in vivo evaluations[J]. J Clin Periodontol, 2012, 39(5): 495-505.
[8] Zhu SJ, Choi BH, Huh JY, et al.A comparative qualitative histological analysis of tissue-engineered bone using bone marrow mesenchymal stem cells, alveolar bone cells, and periosteal cells[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2006, 101(2): 164-169.
[9] 郑文龙, 吴涛, 邹多宏, 等. 犬牙槽骨干细胞与髂骨骨髓干细胞体外成骨能力的比较[J]. 安徽医科大学学报, 2016, 51(9): 1252-1257.
[10] Zhao Y, Chen S, Wang Y, et al.Effect of drying processes on prenylflavonoid content and antioxidant activity of Epimedium koreanum Nakai[J]. J Food Drug Anal, 2018, 26(2): 796-806.
[11] 訾慧, 卢健, 张红梅, 等. 淫羊藿提取物中淫羊藿苷及其代谢产物在骨质疏松模型大鼠肾脏中的分布研究[J]. 中医药信息, 2014, 31(5): 60-63.
[12] 翟远坤, 陈克明, 葛宝丰, 等. 淫羊藿次苷Ⅱ通过激活雌激素信号通路促进骨髓间充质干细胞的成骨性分化[J]. 中国药理学通报, 2011, 27(10): 1451-1457.
[13] Cheng T, Sheng T, Yi Y, et al.Metabolism profiles of icariin in rats using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry and in vitro enzymatic study[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2016, 353(60): 1033-1034
[14] Wang N, Zhang Q, Xin H, et al.Osteoblast cell membrane chromatography coupled with liquid chromatography and time-of-flight mass spectrometry for screening specific active components from traditional Chinese medicines[J]. J Sep Sci, 2017, 40(22): 4311-4319.
[15] Luo G, Xu B, Wang W, et al.Study of the osteogenesis effect of icariside II and icaritin on canine bone marrow mesenchymal stem cells[J]. J Bone Miner Metab, 2018, 36(6): 668-678
[16] Li G, Hu J, Chen H, et al.Enamel matrix derivative enhances the proliferation and osteogenic differentiation of human periodontal ligament stem cells on the titanium implant surface[J]. Organogenesis, 2017, 13(3): 103-113.
[17] Liu H, Zhang N, Liu Y, et al.Effect of human wnt10b transgene overexpression on peri-implant osteogenesis in ovariectomized rats[J]. Hum Gene Ther, 2018, 29(12): 1416-1427
[18] Liu Y, Ming L, Luo H, et al.Integration of a calcined bovine bone and BMSC-sheet 3D scaffold and the promotion of bone regeneration in large defects[J]. Biomaterials, 2013, 34(38): 9998-10006.
[19] Pang XG, Cong Y, Bao NR, et al.Quercetin stimulates bone marrow mesenchymal stem cell differentiation through an estrogen receptor-mediated pathway[J]. Biomed Res Int, 2018, 2018: 4178021.
[20] Zhang L, Jiang G, Zhao X, et al.Dimethyloxalylglycine promotes bone marrow mesenchymal stem cell osteogenesis via Rho/ROCK signaling[J]. Cell Physiol Biochem, 2016, 39(4): 1391-1403.

淫羊藿次苷对犬颌骨间充质干细胞增殖与成骨分化的影响

Effects of Icarisid Ⅱ on proliferation and osteogenic differentiation of alveolar bone-derived stem cells in Beagle dogs

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