Effect of Toll-like receptor 2 agonist Pam3CSK4 on the expression of several cytokines in human dental pulp cells

doi:10.3877/cma.j.issn.1674-1366.2013.05.001

Abstract

Abstract:

Objective

This study was to investigate the effect of Toll-like receptor 2 agonist Pam3CSK4 on the expression of several cytokines in human dental pulp cells （hDPCs）.

Methods

The mRNA expression of IL-6， IL-8， IL-1β and CXCL10， and the protein expression of IL-6， IL-8 in hDPCs with 1 μg/ml Pam3CSK4 at different time points were examined by real-time quantitative PCR and ELISA， respectively. Nuclear location of p65 was investigated by immunofluorescent staining in hDPCs.

Results

The mRNA expression of IL-6 firstly elevated at 4 h， whereas the expression of IL-8，IL-1β and CXCL10 peaked at 8 hours by the incubation with 1 μg/ml Pam3CSK4. IL-6 and IL-8 proteins were clearly expressed in hDPCs culture supernatant， which increased at 4 h and maintained stable at 8 h. Confocal laser scanning microscope showed the positive expression of p65 in nuclear with 1 μg/ml Pam3CSK4 for 75 min， whereas p65 was only observed in cytoplasm in untreated control group.

Conclusions

Activation of TLR2 by Pam3CSK4 induced the expression of several cytokines both at mRNA and protein levels at 4 ～8 h through promoting NF-κB activation in hDPCs， indicating they may be involved in immune regulation of early pulpitis.

Key words: Human dental pulp cells, Toll-like receptor 2, Pam3CSK4, Cytokines, Nuclear factor-κB

Li-li HONG, Yu DU, Qian-ying KONG, Xi WEI. Effect of Toll-like receptor 2 agonist Pam3CSK4 on the expression of several cytokines in human dental pulp cells[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2013, 07(05): 347-351.

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URL: https://zhkqyxyjzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-1366.2013.05.001

https://zhkqyxyjzz.cma-cmc.com.cn/EN/Y2013/V07/I05/347

Figures/Tables 4

References 21

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基因	引物序列
CXCL10	正向：5'-CACCATGAATCAAACTGCGA-3'
	反向：5'-GCTGATGCAGGTACAGCGT-3'
IL-1β	正向：5'-GAAGCTGATGGCCCTAAACA-3'
	反向：5'-AAGCCCTTGCTGTAGTGGTG-3'
IL-6	正向：5'-AGTGAGGAACAAGCCAGAGC-3'
	反向：5'-GTCAGGGGTGGTTATTGCAT-3'
IL-8	正向：5'-TCCTGATTTCTGCAGCTCTGT-3'
	反向：5'-AAATTTGGGGTGGAAAGGTT-3'
GADPH	正向：5'-AAGGTGAAGGTCGGAGTCAA-3'
	反向：5'-AATGAAGGGGTCATTGATGG-3'

基因	引物序列
CXCL10	正向：5'-CACCATGAATCAAACTGCGA-3'
	反向：5'-GCTGATGCAGGTACAGCGT-3'
IL-1β	正向：5'-GAAGCTGATGGCCCTAAACA-3'
	反向：5'-AAGCCCTTGCTGTAGTGGTG-3'
IL-6	正向：5'-AGTGAGGAACAAGCCAGAGC-3'
	反向：5'-GTCAGGGGTGGTTATTGCAT-3'
IL-8	正向：5'-TCCTGATTTCTGCAGCTCTGT-3'
	反向：5'-AAATTTGGGGTGGAAAGGTT-3'
GADPH	正向：5'-AAGGTGAAGGTCGGAGTCAA-3'
	反向：5'-AATGAAGGGGTCATTGATGG-3'

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