Cyclic tensile stress effects on the expression of connective tissue growth factor in human periodontal ligament fibroblasts

doi:10.3969/j.issn.2095-4344.2015.33.008

Abstract

Abstract:

BACKGROUND: Previous studies have found that cyclic tensile stress can induce the proliferation of human periodontal ligament fibroblasts in a certain time.
OBJECTIVE: To observe the effect of cyclic tensile stress on the expression of connective tissue growth factor in human periodontal ligament fibroblasts, and to clarify the role of JNK, p38MAPK, PI3K pathway in the cyclic tensile stress-induced expression of connective tissue growth factor in human periodontal ligament fibroblasts.
METHODS: Human periodontal ligament fibroblasts cultured in vitro were given stimulation of cyclic tensile stress in vitro for 1, 6, 12, 24 hours using multi-channel cell stretch stress loading system (experimental group). Control group was set up and not given any stimulation at the same time. The cells under stress application for 12 hours were given specific inhibitors of JNK, p38MAPK, PI3K respectively at the beginning, and were compared with those with no inhibitors. The concentrations of connective tissue growth factor in the culture supernatant were measured by ELISA. The mRNA expression of connective tissue growth factor in human periodontal ligament fibroblasts was detected by real-time RT-PCR.
RESULTS AND CONCLUSION: Compared with the control group, the expression of connective tissue growthfactor in human periodontal ligament fibroblasts under cyclic tensile stress began to increase at 1 hour, increased manifestly at 6 hours, reached the peak at 12 hours, and then decreased at 24 hours. The expression of connective tissue growth factor was reduced by the specific inhibitors of JNK, while the specific inhibitors of p38 MAPK, PI3K had not the same effect. In a certain time, the cyclic tensile stress can induce a time-dependent increase in mRNA and protein levels of connective tissue growth factor. Along with the extending of time, the expression of connective tissue growth factor begins to decrease. Cyclic tensile stress can regulate expression of connective tissue growth factor in human periodontal ligament fibroblasts via the JNK pathway.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Periodontal Ligament, Fibroblasts, Connective Tissue Growth Factor

CLC Number:

R318

Zhang Yue, Jiang Chang-qing, Yu Xin-juan. Cyclic tensile stress effects on the expression of connective tissue growth factor in human periodontal ligament fibroblasts[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(33): 5284-5288.

References

[1] Shibata M,Shintaku Y,Matsuzaki K,et al. The effect of IL-17 on the production of proinflammatory cytokines and matrix metalloproteinase-1 by human periodontal ligament fibroblasts. Orthod Craniofac Res. 2014;17(1):60-68.
[2] Bauer S, Eisinger K, Wiest R, et al. Connective tissue growth factor level is increased in patients with liver cirrhosis but is not associated with complications or extent of liver injury. Regul Pept.2012;179(1-3): 10-14.
[3] Kono M, Nakamura Y, Suda T, et al. Plasma CCN2 (connective tissue growth factor; CTGF) is a potential biomarker in idiopathic pulmonary fibrosis (IPF). Clin Chim Acta.2011;412(23-24): 2211-2215.
[4] Lu W, Liu S, Zhao Z, et al. The effect of connective tissue growth factor on renal fibrosis and podocyte injury in hypertensive rats.Ren Fail.2014;36(9): 1420-1427.
[5] Jacobson A,Cunningham JL.Connective tissue growth factor in tumor pathogenesis. Fibrogenesis Tissue Repair.2012; 5(Suppl 1 Proceedings of Fibroproliferative disorders: from biochemical analysis to targeted therapiesPetro E Petrides and David Brenner): S8.
[6] Asano M, Kubota S, Nakanishi T, et al. Effect of connective tissue growth factor (CCN2/CTGF) on proliferation and differentiation of mouse periodontal ligament-derived cells. Cell Commun Signal.2005;3: 11.
[7] Nitulescu EA, Craitoiu MM, Banita MI, et al. The involvement of TGF-beta1 and CTGF in regional gingival overgrowth. Rom J Morphol Embryol.2012;53(1): 143-150.
[8] 李松泽,莫水学.牙周组织中结缔组织生长因子的研究进展[J]. 广西医科大学学报, 2010, 27(6): 971-973.
[9] Chu CY, Chang CC, Prakash E, et al. Connective tissue growth factor (CTGF) and cancer progression. J Biomed Sci.2008;15(6): 675-685.
[10] Xiao LW, Yang M, Dong J, et al. Stretch-inducible expression of connective tissue growth factor (CTGF) in human osteoblasts-like cells is mediated by PI3K-JNK pathway. Cell Physiol Biochem.2011;28(2): 297-304.
[11] Yamashiro T, Fukunaga T, Kobashi N, et al. Mechanical stimulation induces CTGF expression in rat osteocytes. J Dent Res.2001;80(2): 461-465.
[12] 段培佳,陈雨雪,陈扬熙.大鼠正畸牙移动张力侧牙周组织中CTGF基因表达的研究[J].口腔医学研究, 2009, 25(4): 433-435.
[13] Sakai Y, Balam TA, Kuroda S, et al. CTGF and apoptosis in mouse osteocytes induced by tooth movement. J Dent Res. 2009;88(4): 345-350.
[14] El-Shewy HM,Sohn M,Wilson P,et al.Low-density lipoprotein induced expression of connective tissue growth factor via transactivation of sphingosine 1-phosphate receptors in mesangial cells. Mol Endocrinol.2012;26(5): 833-845.
[15] Shi L, Chang Y, Yang Y,et al.Activation of JNK signaling mediates connective tissue growth factor expression and scar formation in corneal wound healing. PLoS One.2012;7(2): e32128.
[16] Ko WC, Chen BC, Hsu MJ, et al. Thrombin induced connective tissue growth factor expression in rat vascular smooth muscle cells via the PAR-1/JNK/AP-1 pathway. Acta Pharmacol Sin.2012;33(1): 49-56.
[17] Cabello-Verrugio C, Cordova G, Vial C, et al. Connective tissue growth factor induction by lysophosphatidic acid requires transactivation of transforming growth factor type beta receptors and the JNK pathway. Cell Signal.2011;23(2): 449-457.
[18] 杨建波,李孝生.川芎嗪干预转化生长因子β1诱导肝星状细胞结缔组织生长因子表达中p38丝裂酶原激活蛋白激酶的途径[J]. 中国组织工程研究与临床康复, 2010,14(37): 6877-6881.
[19] 荣琳琳,黄海长,喻陆,等.低氧通过p38丝裂素激活蛋白激酶途径刺激人肾间质成纤维细胞表达结缔组织生长因子[J].北京大学学报:医学版,2005, 37(4): 378-381.
[20] Kim JS, Choi IG, Lee BC, et al. Neuregulin induces CTGF expression in hypertrophic scarring fibroblasts. Mol Cell Biochem.2012;365(1-2): 181-189.
[21] 李彩蓉,蔡飞,黎荣,等.氟伐他汀对醛固酮诱导的系膜细胞SGK1及CTGF表达的影响[J].中国病理生理杂志, 2010, 26(5): 956-960.