matrilin-2和matrilin-4在大鼠牙髓损伤动物模型中的表达
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摘要
目的建立大鼠牙髓损伤修复的动物模型,观察matrilin-2、matrilin-4在正常牙髓及牙髓损伤修复过程中的定位、定性表达过程。方法选择30只雄性wistar大鼠,平均分为5组。随机选取一组作为正常组,其余四组在大鼠左侧上颌第一磨牙咬合面开髓作为实验组。分别于3,7,14,21d灌流固定处死动物并取大鼠上颌左侧第一磨牙标本,经固定、脱钙、切片、HE染色与免疫组化染色在光镜下观察牙髓修复的形态学变化及matrilin-2与matrilin-4在正常牙髓及牙髓损伤修复过程中的定位定性表达。结果在正常牙髓中matrilin-2在整个牙髓组织中呈强阳性表达,matrilin-4仅在成牙本质细胞层中表达。在实验组牙髓中,随着时间的推移,matrilin-2在成牙本质细胞层及相邻牙髓细胞中的表达逐渐减弱,而matrilin-4在该部位的表达逐渐增强。结论 matrilin-2和matrilin-4的表达与牙髓炎症发生发展的程度有关,它们可能参与牙髓损伤修复的过程。
Objective To establish a dental pulp wound healing model of rat and observe the locating and qualitative expression of matrilin-2 and matrilin-4 in normal pulp and wound healing pulp.Methods Thirty Wistar rats were selected for this study,they were divided into 5 groups averagely.One group was selected as control group,the others were experimental groups.These rats were prepared an exposure of pulp on the occlusal surface of the left first malar.Four rats,one in each group,were put to death respectively on the 3rd,7th,14 th and 21 th day after operations.The teeth of each rat were fixed,decalcified and made to sections,which were HE staining and immunohistological staining,then observed the morphology changes as well as the expression of matrilin-2 and matrilin-4 in normal pulp and wound healing pulp under microscope.Results In the control group,matrilin-2 was showed to be strongly positive expression,while matrilin-4 was only positive expressed in odontoblast cell layer.In experimental groups,as time went by,the positive expression of matrilin-2 in odontoblast cell layer and pulp cell nearby gradually decreased,however the positive expression of matrilin-4 was strengthened gradually.Conclusion The expression of matrilin-2 and matrilin-4was connected with the degree of pulp inflammation,they may participate in the process of dental pulp wound healing.
Objective To establish a dental pulp wound healing model of rat and observe the locating and qualitative expression of matrilin-2 and matrilin-4 in normal pulp and wound healing pulp.Methods Thirty Wistar rats were selected for this study,they were divided into 5 groups averagely.One group was selected as control group,the others were experimental groups.These rats were prepared an exposure of pulp on the occlusal surface of the left first malar.Four rats,one in each group,were put to death respectively on the 3rd,7th,14 th and 21 th day after operations.The teeth of each rat were fixed,decalcified and made to sections,which were HE staining and immunohistological staining,then observed the morphology changes as well as the expression of matrilin-2 and matrilin-4 in normal pulp and wound healing pulp under microscope.Results In the control group,matrilin-2 was showed to be strongly positive expression,while matrilin-4 was only positive expressed in odontoblast cell layer.In experimental groups,as time went by,the positive expression of matrilin-2 in odontoblast cell layer and pulp cell nearby gradually decreased,however the positive expression of matrilin-4 was strengthened gradually.Conclusion The expression of matrilin-2 and matrilin-4was connected with the degree of pulp inflammation,they may participate in the process of dental pulp wound healing.
引文
1 Mai JN,Chen CC,Ling JQ.Research progress on Matrilin family.International Journal of Stomatology,2009,36(6):691-697.
2 Klatt AR,Nitsche DP,Kobbe B,et al.Molecular structure,processing,and tissue distribution of matrilin-4.Biol Chem,2001,276:17267-17275.
3 Piecha D,Muratoglu S,Morgelin M,et al.Matrilin-2,a large,oligomeric matrix protein,is expressed by a great variety of cells and forms brillar networks.Biol Chem,1999,274:13353-13361.
4 Szabo E,Lodi C,Korpos E,et al.Expression of matrilin-2in oval cells during rat liver regeneration.Matrix Biol,2007,26:554-560.
5 Ichikawa T,Suenaga Y,Koda T,et al.Delta Np63/BMP-7-dependent expression of matrilin-2 is involved in keratinocyte migration in response to wounding.Biochem Biophys Res Commun,2008,369:994-1000.
6 Ferenc D,Lajos M,va K,et al.Extracellular deposition of matrilin-2 controls the timing of the myogenic program during muscle regeneration.Cell Sci,2014,127(15):3240-3256.
7 Malin D,Sonnenberg-Riethmacher E,Guseva D,et al.The extracellular-matrix protein matrilin 2 participates in peripheral nerve regeneration.Cell Sci,2009,122:1471.
8 Eszter S,Szabolcs F,Zoltán H.Fibrillin-2,tenascin-C,matrilin-2,and matrilin-4 are strongly expressed in the epithelium of human granular and lattice type I corneal dystrophies.Mol Vis,2012,(18):1927-1936.
9 Anna J,Stefan T,Tanja K,et al.Axonally derived matrilin-2 induces proinflammatory responses that exacerbate autoimmune neuroinflammation.Clin Invest,2014,124(11):5042-5056.
10 Wagener R,Ehlen HW,Ko YP,et al.The matrilins–adaptor proteins in the extracellular matrix.FEBS Lett,2005,579:3323-3329.
11 Gronthos S,Mankani M,Brahim J,et al.Postnatal human dental pulp stem cells(DPSCs)in vitro and in vivo.Proc Natl Acad Sci USA,2000,97:13625-13630.
12 Gronthos S,Brahim J,Li W,et al.Stem cell properties of human dental pulp stem cells.Dent Res,2002,81:531-535.
13 Sloan AJ,Smith AJ.Stem cells and the dental pulp:potential roles in dentine regeneration and repair.Oral Dis,2007,13:151-157.
14 Mitsiadis TA,Rahiotis C.Parallels between tooth development and repair:conserved molecular mechanisms following carious and dental injury.Dent Res,2004,83:896-902.
15 Paakkonen V,Vuoristo JT,Salo T.Comparative gene expression profile analysis between native human odontoblasts and pulp tissue.Int Endod,2008,41(3):117-127.
16 Klatt AR,Nitsche DP,Kobbe B,et al.Molecular structure,processing,and tissue distribution of matrilin-4.Biol Chem,2001,276:17267-17275.
17 Lu H,Liu SW,Cui Z,et al.Roles of apoptosis and mitosis in the regeneration of rat parotid gland following ligationinduced atrophy.Journal of Oral Science Research,2015,31(5):444-447.
18 Jegat N,Septier D,Veis A,et al.Short-term effects of amelogenin gene splice products A+4 and A-4 implanted in the exposed rat molar pulp.Head Face Med,2007,3:40.
19 Goldberg M,Smith AJ.Cells and extracellular matrices of dentin and pulp:abiological basis for repair and tissue engineering.Crit Rev Oral Biol Med,2004,15:13-27.
20 Tranasi M,Sberna MT,Zizzari V,et al.Microarray evaluation of age-related changes in human dental pulp.Endod,2009,35:1211-1217.
21 Zhang S,Peng J,Guo Y.Matrilin-2 is a widely distributed extracellular matrix protein and a potential biomarker in the early stage of osteoarthritis in articular cartilage.Biomed Res Int 2014,2014,986127
22 Wang Z,Luo J,Iwamoto S,et al.Matrilin-2 is proteolytically cleaved by ADAMTS-4 and ADAMTS-5.Molecules,2014,19(6):8472-8487.
23 Paakkonen V,Vuoristo JT,Salo T,et al.Comparative gene expression profile analysis between native human odontoblasts and pulp tissue.Int Endod J,2008,41:117-127.
24 Korpos E,Molnar A,Papp P,et al.Expression pattern of matrilins and other extracellular matrix proteins characterize distinct stages of cell differentiation during antler development.Matrix Biol,2005,24:124-135.
2 Klatt AR,Nitsche DP,Kobbe B,et al.Molecular structure,processing,and tissue distribution of matrilin-4.Biol Chem,2001,276:17267-17275.
3 Piecha D,Muratoglu S,Morgelin M,et al.Matrilin-2,a large,oligomeric matrix protein,is expressed by a great variety of cells and forms brillar networks.Biol Chem,1999,274:13353-13361.
4 Szabo E,Lodi C,Korpos E,et al.Expression of matrilin-2in oval cells during rat liver regeneration.Matrix Biol,2007,26:554-560.
5 Ichikawa T,Suenaga Y,Koda T,et al.Delta Np63/BMP-7-dependent expression of matrilin-2 is involved in keratinocyte migration in response to wounding.Biochem Biophys Res Commun,2008,369:994-1000.
6 Ferenc D,Lajos M,va K,et al.Extracellular deposition of matrilin-2 controls the timing of the myogenic program during muscle regeneration.Cell Sci,2014,127(15):3240-3256.
7 Malin D,Sonnenberg-Riethmacher E,Guseva D,et al.The extracellular-matrix protein matrilin 2 participates in peripheral nerve regeneration.Cell Sci,2009,122:1471.
8 Eszter S,Szabolcs F,Zoltán H.Fibrillin-2,tenascin-C,matrilin-2,and matrilin-4 are strongly expressed in the epithelium of human granular and lattice type I corneal dystrophies.Mol Vis,2012,(18):1927-1936.
9 Anna J,Stefan T,Tanja K,et al.Axonally derived matrilin-2 induces proinflammatory responses that exacerbate autoimmune neuroinflammation.Clin Invest,2014,124(11):5042-5056.
10 Wagener R,Ehlen HW,Ko YP,et al.The matrilins–adaptor proteins in the extracellular matrix.FEBS Lett,2005,579:3323-3329.
11 Gronthos S,Mankani M,Brahim J,et al.Postnatal human dental pulp stem cells(DPSCs)in vitro and in vivo.Proc Natl Acad Sci USA,2000,97:13625-13630.
12 Gronthos S,Brahim J,Li W,et al.Stem cell properties of human dental pulp stem cells.Dent Res,2002,81:531-535.
13 Sloan AJ,Smith AJ.Stem cells and the dental pulp:potential roles in dentine regeneration and repair.Oral Dis,2007,13:151-157.
14 Mitsiadis TA,Rahiotis C.Parallels between tooth development and repair:conserved molecular mechanisms following carious and dental injury.Dent Res,2004,83:896-902.
15 Paakkonen V,Vuoristo JT,Salo T.Comparative gene expression profile analysis between native human odontoblasts and pulp tissue.Int Endod,2008,41(3):117-127.
16 Klatt AR,Nitsche DP,Kobbe B,et al.Molecular structure,processing,and tissue distribution of matrilin-4.Biol Chem,2001,276:17267-17275.
17 Lu H,Liu SW,Cui Z,et al.Roles of apoptosis and mitosis in the regeneration of rat parotid gland following ligationinduced atrophy.Journal of Oral Science Research,2015,31(5):444-447.
18 Jegat N,Septier D,Veis A,et al.Short-term effects of amelogenin gene splice products A+4 and A-4 implanted in the exposed rat molar pulp.Head Face Med,2007,3:40.
19 Goldberg M,Smith AJ.Cells and extracellular matrices of dentin and pulp:abiological basis for repair and tissue engineering.Crit Rev Oral Biol Med,2004,15:13-27.
20 Tranasi M,Sberna MT,Zizzari V,et al.Microarray evaluation of age-related changes in human dental pulp.Endod,2009,35:1211-1217.
21 Zhang S,Peng J,Guo Y.Matrilin-2 is a widely distributed extracellular matrix protein and a potential biomarker in the early stage of osteoarthritis in articular cartilage.Biomed Res Int 2014,2014,986127
22 Wang Z,Luo J,Iwamoto S,et al.Matrilin-2 is proteolytically cleaved by ADAMTS-4 and ADAMTS-5.Molecules,2014,19(6):8472-8487.
23 Paakkonen V,Vuoristo JT,Salo T,et al.Comparative gene expression profile analysis between native human odontoblasts and pulp tissue.Int Endod J,2008,41:117-127.
24 Korpos E,Molnar A,Papp P,et al.Expression pattern of matrilins and other extracellular matrix proteins characterize distinct stages of cell differentiation during antler development.Matrix Biol,2005,24:124-135.