炎症微环境下芦丁对牙周膜干细胞成骨分化能力的影响

doi:10.19439/j.sjos.2019.04.004

上海口腔医学 ›› 2019, Vol. 28 ›› Issue (4): 356-361.doi: 10.19439/j.sjos.2019.04.004

炎症微环境下芦丁对牙周膜干细胞成骨分化能力的影响

赵斌, 张云鹏, 徐欣

山东省口腔组织再生重点实验室,山东大学口腔医学院,山东济南 250012

收稿日期:2019-01-30 修回日期:2019-03-13 出版日期:2019-08-25 发布日期:2019-09-23
通讯作者: 徐欣,E-mail：xinxu@sdeu.edu.cn
作者简介:赵斌(1991-),男,硕士研究生,E-mail：1479008539@qq.com
基金资助:
山东省泰山学者工程专项经费 (ts201511106)

Rutin promotes osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment

ZHAO Bin, ZHANG Yun-peng, XU Xin

Shandong Provincial Key Laboratory of Oral Tissue Regeneration; School of Stomatology, Shandong University. Jinan 250012, Shandong Province, China

Received:2019-01-30 Revised:2019-03-13 Online:2019-08-25 Published:2019-09-23

摘要/Abstract

摘要： 目的研究芦丁（rutin）对炎症微环境下牙周膜干细胞成骨分化能力的影响。方法采用有限稀释法分离纯化获得牙周膜干细胞,采用流式细胞术鉴定牙周膜干细胞。以脂多糖（lipopolysaccharide, LPS）刺激牙周膜干细胞,建立体外炎症模型。实验分为4组,第1组使用α-MEM培养基培养牙周膜干细胞,第2组使用含有脂多糖的α-MEM培养基培养牙周膜干细胞,第3组在含有脂多糖的α-MEM培养基加入芦丁培养牙周膜干细胞,第4组使用含有芦丁的α-MEM培养基培养牙周膜干细胞。通过碱性磷酸酶染色、碱性磷酸酶活性测试、茜素红染色、RT-PCR以及蛋白免疫印迹等方法检测成骨分化能力的改变。采用SPSS 17.0软件包对数据进行统计分析。结果 CCK-8和碱性磷酸酶活性测试结果显示,10 μmol/L芦丁对炎症状态下牙周干细胞增殖和成骨分化作用最明显。碱性磷酸酶染色和茜素红染色结果显示,10 μmol/L芦丁可以改善炎症微环境下牙周膜干细胞的成骨分化能力。RT-PCR、蛋白免疫印迹结果显示,芦丁可以增强炎症状态下COL1、ALP、RUNX2等成骨基因和成骨蛋白的表达。结论芦丁可以增强炎症微环境下牙周膜干细胞的成骨分化能力。

关键词: 芦丁, 脂多糖, 牙周膜干细胞, 成骨分化

Abstract: PURPOSE: To explore the effect of rutin on osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment. METHODS: Periodontal ligament stem cells (PDLSCs) were obtained by limited dilution method in vitro. PDLSCs were identified by flow cytometery. Lipopolysaccharide(LPS) was used to stimulate human periodontal ligament stem cells to establish an inflammation model in vitro. The experiment was divided into 4 groups: in group 1, only α-MEM was used to culture PDLSCs; in group 2, α-MEM medium containing LPS was used to culture PDLSCs, in group 3, rutin was added to α-MEM medium containing LPS to PDLSCs; and in group 4, α-MEM medium containing rutin was used to culture PDLSCs. Cell counting kit-8 was used to detect cell proliferation activity. Alkaline phosphatase(ALP) staining, ALP activity test, alizarin red staining, RT-PCR, and Western blot were used to detect the changes of osteogenic differentiation ability. The data were analyzed by SPSS 17.0 software package. RESULTS: The results of CCK-8 and ALP activity analysis showed that rutin at 10 μmol/L could significantly promote the proliferation and differentiation of periodontal stem cells under inflammatory state. ALP staining and alizarin red staining proved that (10 μmol/L) rutin could improve osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment. RT-PCR and Western blot results showed that rutin could enhance the expression of osteogenic genes and proteins such as COL1, ALP, and RUNX2 under inflammatory state. CONCLUSIONS: Rutin can promote osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment.

Key words: Rutin, Lipopolysaccharide, Periodontal ligament stem cells, Osteogenic differentiation

中图分类号:

R781.4

赵斌, 张云鹏, 徐欣. 炎症微环境下芦丁对牙周膜干细胞成骨分化能力的影响[J]. 上海口腔医学, 2019, 28(4): 356-361.

ZHAO Bin, ZHANG Yun-peng, XU Xin. Rutin promotes osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment[J]. Shanghai Journal of Stomatology, 2019, 28(4): 356-361.

参考文献

[1] Chapple IL, Van der Weijden F, Doerfer C, et al. Primary prevention of periodontitis: managing gingivitis[J]. J Clin Periodontol, 2015, 42(Suppl 16): S71-76.
[2] Pihlstrom BL, Michalowicz BS, Johnson NW.Periodontal diseases[J]. Lancet, 2005, 366(9499):1809-1820.
[3] Chen FM, Sun HH, Lu H, et al.Stem cell-delivery therapeutics for periodontal tissue regeneration[J]. Biomaterials, 2012, 33(27): 6320-6344.
[4] Ji K, Liu Y, Lu W, et al.Periodontal tissue engineering with stem cells from the periodontal ligament of human retained deciduous teeth[J]. J Periodontal Res, 2013, 48(1): 105-116.
[5] Zhu B, Liu W, Liu Y, et al.Jawbone microenvironment promotes periodontium regeneration by regulating the function of periodontal ligament stem cells[J]. Sci Rep, 2017, 7: 40088.
[6] Liu N, Shi S, Deng M, et al.High levels of β-catenin signaling reduce osteogenic differentiation of stem cells in inflammatory microenvironments through inhibition of the noncanonical Wnt pathway[J]. J Bone Miner Res, 2011, 26(9): 2082-2095.
[7] Ross JA, Kasum CM.Dietary flavonoids: bioavailability, metabolic effects, and safety[J]. Annu Rev Nutr, 2003, 22: 19-34.
[8] Xu J, Jia YY, Chen SR, et al.(E)-1-(4-ethoxyphenyl)-3-(4-nitrophenyl)-prop-2-en-1-one suppresses LPS-induced inflammatory response through inhibition of NF-κB signaling pathway[J]. Int Immunopharmacol, 2013, 15(4): 743-751.
[9] Li Q, Qiu YL, Mao M, et al.Antioxidant mechanism of rutin on hypoxia-induced pulmonary arterial cell proliferation[J]. Molecules, 2014, 19(11): 19036-19049.
[10] Hyun H, Park H, Jeong J, et al.Effects of watercress containing rutin and rutin alone on the proliferation and osteogenic differentiation of human osteoblast-like MG-63 cells[J]. Korean J Physiol Pharmacol, 2014, 18(4): 347-352.
[11] Rassi CM, Lieberherr M, Chaumaz G, et al.Modulation of osteoclastogenesis in porcine bone marrow cultures by quercetin and rutin[J]. Cell Tissue Res, 2005, 319(3): 383-393.
[12] Li C, Li B, Dong Z, et al.Lipopolysaccharide differentially affects the osteogenic differentiation of periodontal ligament stem cells and bone marrow mesenchymal stem cells through Toll-like receptor 4 mediated nuclear factor κB pathway[J]. Stem Cell Res Ther, 2014, 5(3): 67.
[13] Li C, Wang X, Tan J, et al.The immunomodulatory properties of periodontal ligament stem cells isolated from inflamed periodontal granulation[J]. Cells Tissues Organs, 2014, 199(4): 256-265.
[14] Maeda H, Tomokiyo A, Fujii S, et al.Promise of periodontal ligament stem cells in regeneration of periodontium[J]. Stem Cell Res Ther, 2011, 2(4): 33.
[15] De Wilde A, Lieberherr M, Colin C, et al.A low dose of daidzein acts as an ERbeta - selective agonist in trabecular osteoblasts of young female piglets[J]. J Cell Physiol, 2004, 200(2): 253-262.
[16] Ming LG, Chen KM, Xian CJ, et al.Functions and actions mechanisms of flavonoids genistein and icariin in regulating bone remodeling[J]. J Cell Physiol, 2014, 228(3): 513-521.
[17] Zhang JF, Li G, Meng CL, et al.Total flavonoids of Herba Epimedii improves osteogenesis and inhibits osteoclastogenesis of human mesenchymal stem cells[J]. Phytomedicine, 2009, 16(6-7): 521-529.
[18] Fu JH, Zheng YQ, Li P, et al.Hawthorn leaves flavonoids decreases inflammation related to acute myocardial ischemia/reperfusion in anesthetized dogs[J]. Chin J Integr Med, 2013, 19(8): 582-588.
[19] González R, Ballester I, López-Posadas R, et al.Effects of flavonoids and other polyphenols on inflammation[J]. Crit Rev Food Sci Nutr, 2011, 51(4): 331-362.
[20] Comalada M, Camuesco D, Sierra S, et al.In vivo quercitrin anti-inflammatory effect involves release of quercetin, which inhibits inflammation through down-regulation of the NF-kappaB pathway[J]. Eur J Immunol, 2010, 35(2): 584-592.
[21] Dacic S, Kalajzic I, Visnjic D, et al.Col1a1-driven transgenic markers of osteoblast lineage progression[J]. J Bone Miner Res, 2010, 16(7):1228-1236.
[22] Deng Y, Wu S, Zhou H, et al.Effects of a miR-31, Runx2, and Satb2 regulatory loop on the osteogenic differentiation of bone mesenchymal stem cells[J]. Stem Cells Dev, 2013, 22(16): 2278-2286.
[23] Perk AA, Shatynska-Mytsyk I, Gerçek YC, et al.Rutin mediated targeting of signaling machinery in cancer cells[J]. Cancer Cell Int, 2014, 14(1): 124.
[24] Arjumand W, Seth A, Sultana S.Rutin attenuates cisplatin induced renal inflammation and apoptosis by reducing NFκB, TNF-α and caspase-3 expression in wistar rats[J]. Food Chem Toxicol, 2011, 49(9): 2013-2021.
[25] Park SE, Sapkota K, Choi JH, et al.Rutin fromdendropanax morbiferaleveille protects human dopaminergic cells against rotenone induced cell injury through inhibiting JNK and p38 MAPK signaling[J]. Neurochem Res, 2014, 39(4): 707-718.

炎症微环境下芦丁对牙周膜干细胞成骨分化能力的影响

Rutin promotes osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment

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