Effects of substance P on mineralization markers and heme oxygenase-1 Expression in human immortalized periodontal ligament cells
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
Substance P (SP) is known to be expressed in the nerve fibers of dental pulp and periodontal tissues. It was recently reported that SP expression increased in response to orthodontic force. In the present study, we investigated the effect of SP on expression of mineralization markers and heme oxygenase-1 (HO-1) in human immortalized periodontal ligament (IPDL) cells. Cell viability was measured using a 3,4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) assay. The expression of mineralization markers, including alkaline phosphatase (ALP), osteonectin (ON) and bone sialoprotein (BSP), and heme oxygenase-1 (HO-1) was assessed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. SP did not significantly change human IPDL cell viability, with the exception of the 24 hour treatment group. Treatment of human IPDL cells with 10 -^10 to 10 -⁴ M SP upregulated mineralization marker and HO-1 expression in a time- and concentration-dependent manner. Our results suggest that SP may modulate osteoblastic cell differentiation of human IPDL cells through a mechanism involving HO-1 expression.
Abstract
Substance P (SP) is known to be expressed in the nerve fibers of dental pulp and periodontal tissues. It was recently reported that SP expression increased in response to orthodontic force. In the present study, we investigated the effect of SP on expression of mineralization markers and heme oxygenase-1 (HO-1) in human immortalized periodontal ligament (IPDL) cells. Cell viability was measured using a 3,4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) assay. The expression of mineralization markers, including alkaline phosphatase (ALP), osteonectin (ON) and bone sialoprotein (BSP), and heme oxygenase-1 (HO-1) was assessed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. SP did not significantly change human IPDL cell viability, with the exception of the 24 hour treatment group. Treatment of human IPDL cells with 10 -^10 to 10 -⁴ M SP upregulated mineralization marker and HO-1 expression in a time- and concentration-dependent manner. Our results suggest that SP may modulate osteoblastic cell differentiation of human IPDL cells through a mechanism involving HO-1 expression.
참고문헌 (30)
- Basdra EK, "Osteoblast-like properties of human periodontal ligament cells : an in vitro analysis", European Journal of Orthodontics, 19, pp.615~621 (1997)
- Bjurholm A, "Neuroendocrine regulation of cyclic AMP formation in osteoblastic cell lines (UMR-106-01, ROS 17", 2.8, MC3T3-E1, ans Saos2) and primary bone cells, 1992, pp.7, 1011~1019
- Cho MI, "In vitro formation of mineralized nodules by periodontal ligament cells from the rat", Calcified Tissue International, 50, pp.459~467 (1992)
- Choi AM, "Heme oxygenase-1: function, regulation, and implication of a novel stress-inducible protein in oxidant-induced lung injury", American Journal of Respiratory Cell and Molecular Biology, 15, pp.9~19 (1996)
- Goto T, "Tachykinins and tachykinin receptors in bone", Microscopy Research and Technique, 58, pp.91~97 (2002)
- Hayashi K, "Involvement of nitric oxide in orthodontic tooth movement in rats", American Journal of Orthodontics and Dentofacial Orthopedics, 122, pp.306~309 (2002)
- Imai S, "Neuronal regulation of bone metabolism and anabolism: calcitonin gene-related peptide-, substance P-, and tyrosine hydroxylase-containing nerves and the bone", Microscopy Research and Technique, 58, pp.61~69 (2002)
- Irie K, "Calcitonin generelated peptide (CGRP)-containing nerve fibers in bone tissue and their involvement in bone remodeling", Microscopy Research and Technique, 58, pp.85~90 (2002)
- Jeon IS, "An immunohistochemical study on substance P and calcitonin gene-related peptide in the experimental tooth movement of rat", Journal of WonKwang Dental Research, 7, pp.63~84 (1997)
- Kojima T, "Substance P stimulates release of RANKL via COX-2 expression in human dental pulp cells", Inflammation Research, 55, pp.78~84 (2006)
- Kvinnsland I, "Effect of traumatic occlusion on CGRP and SP immunoreactive nerve fiber morphology in rat molar pulp and periodontium", Histochemistry, 97, pp.111~120 (1992)
- Li MH, "Peroxynitrite induces HO-1 expression via PI3K", Akt-dependent activation of NF-E2-related factor 2 in PC12 cells, 2006, pp.41, 1079~1091
- Long P, "Signaling by mechanical strain involves transcriptional regulation of proinflammatory genes in human periodontal ligament cells in vitro", Bone, 30, pp.547~552 (2002)
- Lutherman J, "Immunohistochemical study of neuronal markers in human gingiva with phenytoin-induced overgrowth", Scandinavian Journal of Dental Research, 96, pp.339~346 (1988)
- Matayoshi T, "Neuropeptide substance P stimulates the formation of osteoclasts via synovial fibroblastic cells", Biochemical and Biophysical Research Communications, 327, pp.756~764 (2005)
- Mizuno M, "Bone sialoprotein (BSP) is a cucial factor for the expression of osteoblastic phenotypes of bone marrow cells cultured on type I collagen matrix", Calcified Tissue International, 66, pp.388~396 (2000)
- Mori T, "Substance P regulates the function of rabbit cultured osteoclast; increase of intracellular free calcium concentration and enhanced bone resorption", Biochemical and Biophysical Research Communications, 262, pp.418~422 (1999)
- Nicolay OF, "Substance P immunoreactivity in periodontal tissues during orthodontic tooth movement", Bone and Mineral, 11, pp.19~29 (1990)
- Nohutcu RM, "Expression of extracellular matrix proteins in human periodontal ligament cells during mineralization in vitro", Journal of Periodontology, 68, pp.320~327 (1997)
- Nojima N, "Fibroblastic cells derived from bovine periodontal ligaments have the phenotypes of osteoblasts", Journal of Periodontal Research, 25, pp.179~185 (1990)
- Norevall LI, "Main sensory neuropeptides, but not VIP and NPY, are involved in bone remodeling during orthodontic tooth movement in the rat", Annals of the New York Academy of Sciences, 865, pp.353~359 (1998)
- Norton LA, "A methodical study of shape changes in human oral cells perturbed by a stimulated orthodontic strain in vitro", Archives of Oral Biology, 40, pp.863~872 (1995)
- Oh JH, "Effects of substance P and VIP on the alkaline phosphatase activity and the gene expression of bone proteins in osteoblast-like cells", Journal of Oral Biology, 19, pp.33~39 (1995)
- Otterbein L, "Hemoglobin provides protection against lethal endotoxemia in rats: the role of heme oxygenase-1", American Journal of Respiratory Cell and Molecular Biology, 13, pp.595~601 (1995)
- Pi SH, "Differential expression of periodontal ligament-specific markers and osteogenic differentiation in HPV16-immortalized human gingival fibroblasts and periodontal ligament cells", Journal of Periodontal Research, 42, pp.104~113 (2007)
- Sohn SJ, "Substance P upregulates osteoclastogenesis by activating nuclear factor kappa B in osteoclast precursors", Acta Oto-Laryngologica, 125, pp.130~133 (2005)
- Termine JD, "Mineral and collagen binding proteins of fetal calf bone", Journal of Biological Chemistry, 256, pp.10403~10408 (1981)
- Terry CM, "TNF-α and IL-1α induce heme oxygenase-1 via protein kinase C, Ca2+ and phospholipase A2 in endothelial cells", American Journal of Physiology - Heart and Circulatory Physiology, 276, pp.1493~1501 (1999)
- Vile GF, "Heme oxygenase-1 mediates an adaptive response to oxidative stress in human skin fibroblasts", Proceedings of the National Academy of Sciences of the United States of America, 91, pp.2607~2610 (1994)
- Zwerina J, "Heme oxygenase 1 (HO-1) regulates osteoclastogenesis and bone resorption", FASEB Journal, 19, pp.2011~2013 (2005)