The effects of Nicotine on the expression of lncRNA NEAT1 in human periodontal ligament cells / 医学研究生学报

ABSTRACT

Objective Several physiological reactions of human periodontal ligament cells (hPDLCs) are effected by a varie-ty of long-chain non-coding RNA , and nicotine as the major risk factor of smoking-related periodontitis , would lead to many cytokines changes in hPDLCs .The aim of the study was to explore the effect of nicotine on the expression of lncRNA NEAT 1 and IL 8 in the nucleus of hPDLCs. Methods We cultured the hPDLCs in vitro and used the 4th generation cells for subsequent experiments .hPDLCs were randomly divided into 4 groups:control group , nicotine group ,α-BTX group, nicotine+α-BTX group.Control group:hPDLCs without any ir-ritation;nicotine group: 10-5 mol/L nicotine stimulated hPDLCs for 4h;α-BTX group:10-8 mol/Lα-BTX stimulated hPDLCs; nicotine+α-BTX group:10-5 mol/L nicotine+10-8 mol/Lα-BTX stimulated hPDLCs .Real-time quantitative PCR was taken to detect the expres-sion level of NEAT1 and downstream IL 8 mRNA.In addition, the protein expression of IL-8 was tested by ELISA. Results After we primary cultured the periodontal ligament tissue for 3 weeks, the cells around the tissue were arranged radially .After a stable passage, microscopic observation showed that hPDLCs were long spindle-shape or spindle-shape with clear contour and adherent growth .Com-pared with nicotine group , the expression of NEAT1-1, NEAT1-2, IL-8 mRNA in hPDLCs in other three groups were all decreased ( P0.05). Conclusion Nicotine can promote the expression of NEAT1 and IL-8 in hPDLCs, which suggets that NEAT 1 may be involved in the development of smoking-related periodontitis by promoting early inflammatory reaction .