体外张应力对人牙周膜细胞串珠素表达的影响

上海口腔医学 ›› 2014, Vol. 23 ›› Issue (6): 675-680.

体外张应力对人牙周膜细胞串珠素表达的影响

王奕¹, 孙超凡¹, 陈利娇¹, 王彦亮², 胡荣党¹

1.温州医科大学附属口腔医院正畸科,2.口腔颌面外科,浙江温州 325027

收稿日期:2014-05-04 修回日期:2014-07-12 出版日期:2014-12-20 发布日期:2015-01-08
通讯作者: 胡荣党,Tel： 0577-88066080,E-mail:hurongdang@hotmail.com E-mail:wangyi.ortho@gmail.com
作者简介:王奕（1986-）,男,硕士研究生
基金资助:
浙江省大学生科技创新活动计划（新苗人才计划）项目（2013R413044）; 温州市科技局项目（Y20120228）

Effect of tensile strain on perlecan expression of human periodontal ligament cells

WANG Yi, SUN Chao-fan, CHEN Li-jiao, WANG Yan-liang, HU Rong-dang

1.Department of Orthodontics,
2. Department of Oral and Maxillofacial Surgery, School of Stomatology, Wenzhou Medical University. Wenzhou 325027, Zhejiang Province, China

Received:2014-05-04 Revised:2014-07-12 Online:2014-12-20 Published:2015-01-08

摘要/Abstract

摘要： 目的：观察体外周期性张应力对人牙周膜细胞（human periodontal ligament cells, hPDLCs）串珠素的表达影响,探讨应力作用下牙周组织改建的分子机制。方法：采用酶消法分离培养hPDLC,通过应力加载仪对细胞施以12% elongation、1 Hz的单轴张应力。加力时长分别为0、12、24、48 h。然后通过实时定量PCR和ELISA方法分别检测张应力下不同时间点串珠素基因和蛋白的表达变化。采用SPSS19.0软件包对数据进行统计学分析。结果：张应力加载后,串珠素mRNA在加力的前12 h内表达水平出现短暂升高,但无显著差异。随着加力时间延长,mRNA表达持续下降,48 h后达到最低水平,至对照组的0.28±0.05 (P<0.05);而细胞基质内的串珠素蛋白表达则随着加力时间一直下降,48 h后从（14.03±0.71） pg/mL（对照组）降低到最低水平（11.06±0.15） pg/mL,差异显著（P<0.05）。结论：张应力刺激可下调人牙周膜细胞串珠素基因及蛋白表达,下降趋势与时间具有相关性,提示串珠素可能在牙周膜对机械力刺激的反应中发挥作用。

关键词: 张应力, 人牙周膜细胞, 串珠素

Abstract: PURPOSE: To investigate the perlecan expression of human periodontal ligament cells (hPDLCs) under cyclic tensile strain in vitro, and learn the molecular mechanism of periodontal remolding during tooth movement. METHODS: hPDLCs isolated by enzyme digestion were loaded with 12% elongation, 1 Hz of uniaxial tensile strain for 12, 24 and 48 h. The unloaded cells were used as control. Real-time PCR and enzyme-linked immune sorbent assay (ELISA) were applied to analyze the mRNA and protein expression of perlecen in each sample respectively. The data was analyzed with SPSS 19.0 software package. RESULTS: Within 12 h, mRNA expression was transiently elevated, but no significant difference was detected compared with the control. After 12 h, the mRNA expression was significantly decreased. It would decreased to (0.28±0.049) at lowest level of control at lowest level at 48 h (P<0.05). The protein expression of perlecan was time-dependently decreased. Specifically, it was downregulated from (14.03±0.71) pg/mL (control) to (11.06±0.15) pg/mL at lowest level at 48 h (P<0.05). CONCLUSIONS: Tensile strain time-dependently down-regulates perlecan expression, indicating perlecan may play a pivot role in PDLCS responding to mechanical loading in vitro.

Key words: Supported, by, Xin, Miao, Talent, Program, of, Zhejiang, Province, (2013R423044), and, Research, Project, of, Wenzhou, Science, and, Technology, Department, (Y20120228).

中图分类号:

R781.4

王奕, 孙超凡, 陈利娇, 王彦亮, 胡荣党. 体外张应力对人牙周膜细胞串珠素表达的影响[J]. 上海口腔医学, 2014, 23(6): 675-680.

WANG Yi¹, SUN Chao-fan¹, CHEN Li-jiao¹, WANG Yan-liang², HU Rong-dang¹. Effect of tensile strain on perlecan expression of human periodontal ligament cells[J]. Shanghai Journal of Stomatology, 2014, 23(6): 675-680.

参考文献

[1] Henneman S, Von den Hoff JW, Maltha JC. Mechanobiology of tooth movement [J]. Eur J Orthod, 2008, 30(3): 299-306.
[2] Magan A, Ripamonti U. Biological aspects of periodontal tissue regeneration: cementogenesis and the induction of Sharpey’s fibres [J]. SADJ, 2013, 68(7): 304-306, 308-312, 314 passim.
[3] Liu M, Dai J, Lin Y, et al. Effect of the cyclic stretch on the expression of osteogenesis genes in human periodontal ligament cells [J]. Gene, 2012, 491(2): 187-193.
[4] Whitelock JM, Melrose J, Iozzo RV. Diverse cell signaling events modulated by perlecan [J]. Biochemistry, 2008, 47(43): 11174-11183.
[5] Jha AK, Yang W, Kirn-Safran CB, et al. Perlecan domain I-conjugated, hyaluronic acid-based hydrogel particles for enhanced chondrogenic differentiation via BMP-2 release [J]. Biomaterials, 2009, 30(36): 6964-6975.
[6] Whitelock JM, Murdoch AD, Iozzo RV, et al. The degradation of human endothelial cell-derived perlecan and release of bound basic fibroblast growth factor by stromelysin, collagenase, plasmin, and heparanases[J]. J Biol Chem, 1996, 271(17): 10079-10086.
[7] Schmidtchen A, Carlstedt I, Malmström A, et al. Inventory of human skin fibroblast proteoglycans. Identification of multiple heparan and chondroitin/dermatan sulphate proteoglycans [J]. Biochem J, 1990, 265(1): 289-300.
[8] Larjava H, Hakkinen L, Rahemtulla F. A biochemical analysis of human periodontal tissue proteoglycans[J].Biochem J,1992,284 (Pt 1):267-274.
[9] 李玉珍, 刘秀华, 蔡莉蓉. 串珠素表达下调参与低氧对大鼠心肌微血管内皮细胞增殖的抑制作用 [J]. 生理学报, 2007, 59(2): 221-226.
[10] Baker AB, Ettenson DS, Jonas M, et al. Endothelial cells provide feedback control for vascular remodeling through a mechanosensitive autocrine TGF-beta signaling pathwayc [J]. Circ Res, 2008, 103(3): 289-297.
[11] Natali AN, Pavan PG, Scarpa C. Numerical analysis of tooth mobility: formulation of a non-linear constitutive law for the periodontal ligament [J]. Dent Mater, 2004, 20(7): 623-629.
[12] Lee RT, Yamamoto C, Feng Y, et al. Mechanical strain induces specific changes in the synthesis and organization of proteoglycans by vascular smooth muscle cells [J]. J Biol Chem, 2001, 276(17): 13847-13851.
[13] 李永明, 冯雪, 林珠, 等. 周期性牵张力对人牙周膜细胞MMP-3及TIMP-1表达的影响[J]. 临床口腔医学杂志, 2004, 20(4): 195-198.
[14] 李永明, 唐林, 张晓东, 等. 牵张应力对人牙周膜细胞MMP-13/TIMP-1表达的影响及其信号转导途径研究 [J]. 医用生物力学, 2010, 25(6): 422-427.
[15] Leonardi R, Talic NF, Loreto C. MMP-13 (collagenase 3) immunolocalisation during initial orthodontic tooth movement in rats [J]. Acta Histochem, 2007, 109(3): 215-220.
[16] Wang B, Lai X, Price C, et al. Perlecan-containing pericellular matrix regulates solute transport and mechanosensing within the osteocyte lacunar-canalicular system[J]. J Bone Miner Res, 2014, 29(4): 878-891.
[17] Lowe DA, Lepori-Bui N, Fomin PV, et al. Deficiency in perlecan/HSPG2 during bone development enhances osteogenesis and decreases quality of adult bone in mice [J]. Calcif Tissue Int, 2014, 95(1): 29-38.
[18] Reiland J, Kempf D, Roy M, et al. FGF2 binding, signaling, and angiogenesis are modulated by heparanase in metastatic melanoma cells [J]. Neoplasia, 2006, 8(7): 596-606.
[19] Elkin M, Ilan N, Ishai-Michaeli R, et al. Heparanase as mediator of angiogenesis: mode of action [J]. FASEB J, 2001, 15(9): 1661-1663.
[20] Takayama S, Murakami S, Nozaki T, et al. Expression of receptors for basic fibroblast growth factor on human periodontal ligament cells [J]. J Periodontal Res, 1998, 33(6): 315-322.