p38 丝裂原活化蛋白激酶在炎症微环境作用下对牙周膜干细胞成骨分化的影响

doi:10.3881/j.issn.1000-503X.2015.01.001

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2015, Vol. 37 ›› Issue (1): 1-7.doi: 10.3881/j.issn.1000-503X.2015.01.001

• Orginal Article • Next Articles

Effects of p38 Mitogen-activated Protein Kinase on Osteogenic Differentiation of Human Periodontal Ligament Stem Cells in Inflammatory Microenvironment

Jia NIE¹, Bo ZHANG², Bin GU³, Na LIU⁴()

¹Department of Stomatology,the First Affiliated Hospital of the PLA General Hospital,Beijing 100000,China.
²Department of Stomatology,the Military General Hospital of Beijing PLA,Beijing 100700,China.
³Department of Geriatric Dentistry,
⁴Institute of Stomatology,the PLA General Hospital,Beijing 100853,China

Received:2014-09-12 Online:2015-02-28 Published:2015-02-12
Supported by:
Supported by the National Natural Sciences Foundation of China(31200741),the China Postdoctoral Science Foundation(2013M532108),and the Beijing Nova program(Z14144000180000)

Abstract

Abstract: ObjectiveTo investigate the expression of mitogen-activated protein kinase(MAPK)in the chronic periodontitis tissue-derived in the periodontal ligament stem cells(PDLSCs)and explore its effect on the osteogenic differentiation of human PDLSCs in inflammatory microenvironment. MethodsPDLSCs were obtained from human healthy individuals(H-PDLSCs)and patients with periodontitis(P-PDLSCs). The tumor necrosis factor(TNF)-α and interleukin(IL)-1β secretion and mRNA expression levels of H-PDLSCs and P-PDLSCs were detected using enzyme-linked immunosorbent assay and real-time quantitative PCR. Immunofluorescence staining was used for determining the protein levels of p38 in PDLSCs. The levels of p38 and p-p38 following culture in osteogenic medium for 7 d of H-PDLSCs and P-PDLSCs were detected using Western blotting.After the PDLSCs were stimulated with SB-203580,the p38 MAPK specific inhibitor,for 30 min and then in osteogenic induction process for 7 days,the expression levels of the osteogenic gene Runx2 and alkaline phosphatase(ALP)were determined by real-time quantitative PCR,and bone formation ability of PDLSCs was tested by alizarin red(AR)staining. ResultsThe secretions of TNF-α and IL-1β were significantly higher in P-PDLSCs compared with H-PDLSCs(68.80±6.70 vs. 34.10±3.07,P=0.001;57.10±4.23 vs. 26.90±2.58,P=0.000). The same trend was seen in the gene expression levels of both TNF-α and IL-1β in PDLSCs(P_TNF-α=0.011,P _IL-1β=0.009). p38 was more strongly induced in P-PDLSCs cells than in H-PDLSCs.The basal level of p38 in H-PDLSCs was lower than that in P-PDLSCs cells cultured in the basic medium. However,the level of p-p38 was increased in H-PDLSCs than in P-PDLSCs under osteogenic condition. Treatment of PDLSCs with SB-203580 and then cultures under osteogenic differentiation lead to significantly decreased expressions of Runx2 and ALP in both H-PDLSCs and P-PDLSCs(H-PDLSCs:P_Runx2=0.044,P_ALP=0.036;P-PDLSCs:P_Runx2=0.017,P_ALP=0.004). Conclusionsp38 MAPK is involved in the inflammatory response of PDLSCs in the chronic inflammatory microenvironment. The inhibition of p38 by SB-203580 also remarkably suppresses the osteogenic differentiation of PDLSCs in a chronic inflammatory microenvironment.

Key words: periodontal ligament stem cells, p38 mitogen activated protein kinases, inflammation, osteogenic differentiation

CLC Number:

R780.2

Jia NIE, Bo ZHANG, Bin GU, Na LIU. Effects of p38 Mitogen-activated Protein Kinase on Osteogenic Differentiation of Human Periodontal Ligament Stem Cells in Inflammatory Microenvironment[J].Acta Academiae Medicinae Sinica, 2015, 37(1): 1-7.

Figures/Tables 4

Table 1

Fig 1

Fig 2

Fig 3

References 18

[1]	Seo BM,Miura M,Gronthos S,et al.Investigation of multipotent postnatal stem cells from human periodontal ligament[J]. Lancet,2004,364(9429):149-155.
[2]	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.
[3]	Kong X,Liu Y,Ye R,et al.GSK3β is a checkpoint for TNF-α-mediated impaired osteogenic differentiation of mesenchymal stem cells in inflammatory microenvironments[J]. Biochim Biophys Acta,2013,1830(11):5119-5129.
[4]	Park EJ,Park SW,Kim HJ,et al.Dehydrocostuslactone inhibits LPS-induced inflammation by p38MAPK-dependent induction of hemeoxygenase-1 in vitro and improves survival of mice in CLP-induced sepsis in vivo[J].Int Immunopharmacol,2014,22(2):332-340.
[5]	Rossa C Jr,Liu M,Patil C,et al.MKK3/6-p38 MAPK negatively regulates murine MMP-13 gene expression induced by IL-1beta and TNF-alpha in immortalized periodontal ligament fibroblasts[J]. Matrix Biol,2005,24(7):478-488.
[6]	Zhou Y,Guan X,Wang H,et al.Hypoxia induces osteogenic/angiogenic responses of bone marrow-derived mesenchymal stromal cells seeded on bone-derived scaffolds via ERK1/2 and p38 pathways[J]. Biotechnol Bioeng,2013,110(6):1794-1804.
[7]	Li XD,Liu ZY,Chang B,et al.Panax notoginseng saponins promote osteogenic differentiation of bone marrow stromal cells through the ERK and P38 MAPK signaling pathways[J]. Cell Physiol Biochem,2011,28(2):367-376.
[8]	Liao QC,Xiao ZS,Qin YF,et al.Genistein stimulates osteoblastic differentiation via p38 MAPK-Cbfa1 pathway in bone marrow culture[J]. Acta Pharmacol Sin,2007,28(10):1597-1602.
[9]	Armitage GC,Wu Y,Wang HY,et al.Low prevalence of a periodontitis-associated interleukin-1 composite genotype in individuals of Chinese heritage[J]. J Periodontol,2000,71(2):164-171.
[10]	Arana-Argáez VE,Delgado-Rizo V,Pizano-Martínez OE,et al.Inhibitors of MAPK pathway ERK1/2 or p38 prevent the IL-1β-induced up-regulation of SRP72 autoantigen in Jurkat cells[J]. J Biol Chem,2010,285(43):32824-32833.
[11]	Araújo AA,Souza TO,Moura LM,et al.Effect of telmisartan on levels of IL-1,TNF-α,down-regulated COX-2,MMP-2,MMP-9 and RANKL/RANK in an experimental periodontitis model[J]. J Clin Periodontol,2013,40(12):1104-1011.
[12]	Shu L,Guan SM,Fu SM,et al.Estrogen modulates cytokine expression in human periodontal ligament cells[J]. J Dent Res,2008,87(2):142-147.
[13]	Reddi D,Brown SJ,Belibasakis GN.Porphyromonas gingivalis induces RANKL in bone marrow stromal cells:involvement of the p38 MAPK[J]. Microb Pathog,2011,51(6):415-420.
[14]	Zhou FH,Foster BK,Zhou XF,et al.TNF-alpha mediates p38 MAP kinase activation and negatively regulates bone formation at the injured growth plate in rats[J]. J Bone Miner Res,2006,21(7):1075-1088.
[15]	Hess K,Ushmorov A,Fiedler J,et al.TNFα promotes osteogenic differentiation of human mesenchymal stem cells by triggering the NF-κB signaling pathway[J]. Bone,2009,45(2):367-376.
[16]	Chang J,Sonoyama W,Wang Z,et al.Noncanonical Wnt-4 signaling enhances bone regeneration of mesenchymal stem cells in craniofacial defects through activation of p38 MAPK[J]. J Biol Chem,2007,282(42):30938-30948.
[17]	Liu G,Vijayakumar S,Grumolato L,et al.Canonical Wnts function as potent regulators of osteogenesis by human mesenchymal stem cells[J]. J Cell Biol,2009,185(1):67-75.
[18]	Artigas N,Ureña C,Rodríguez-Carballo E,et al.Mitogen-activated protein kinase(MAPK)-regulated interactions between Osterix and Runx2 are critical for the Transcriptional Osteogenic Program[J]. J Biol Chem,2014,289(39):27105-27117.

基因Gene	引物序列Primer sequence
肿瘤坏死因子-α Tumor necrosis factor-α	正向Forward:5’-AACTCGAGTGACAAGCCCGTAG-3’	反向Reverse:5’-GTACCAGTTGGTTGTCTTTGA-3’
白细胞介素-1β Interleukin-1β	正向Forward:5’-CCGTGCCTACGAACATGTC-3’	反向Reverse:5’-CACACAGAAGCTCATCGGAG-3’
成骨转录因子 Runx2	正向Forward:5’-CCCGTGGCCTTCAAGGT-3’	反向Reverse:5’-CGTTACCCGCCATGACAGTA-3’
碱性磷酸酶 Alkaline phosphatase	正向Forward:5’-GGACCATTCCCACGTCTTCAC-3’	反向Reverse:5’-CCTTGTAGCCAGGCCCATTG-3’
β-肌动蛋白 β-actin	正向Forward:5’-TGGCACCCAGCACAATGAA-3’	反向Reverse:5’-CTAAGTCATAGTCCGCCTAGAAGCA-3’

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Effects of p38 Mitogen-activated Protein Kinase on Osteogenic Differentiation of Human Periodontal Ligament Stem Cells in Inflammatory Microenvironment

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