The effects of Nicotine on the expression of lncRNA NEAT1 in human periodontal ligament cells
10.16571/j.cnki.1008-8199.2017.03.004
- VernacularTitle:尼古丁对人牙周膜细胞长链非编码RNA NEAT1表达的影响
- Author:
Shuai YUAN
;
Yang DU
;
Lulu WANG
;
Zhifei ZHOU
;
Xingan WU
;
Xiaojing WANG
- Keywords:
Human periodontal ligament cells;
Nicotine;
NEAT1;
Inflammatory reaction;
IL-8
- From:
Journal of Medical Postgraduates
2017;30(3):240-244
- CountryChina
- Language:Chinese
-
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 ( P<0.05) .The expression level of IL-8 protein in the supernatant of the nicotine group was significantly higher than the control group , about 4.8 times higher than the control group ( P<0.05) , while theα-BTX group and nicotine+α-BTX group were not significantly dif-ferent from the control group (P>0.05). 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 .
