A preliminary study on the autophagy level of human periodontal ligament cells regulated by nicotine

doi:10.7518/hxkq.2017.02.017

Abstract

Abstract:

Objective To explore the effect of nicotine on the autophagy level of human periodontal ligament cells (hPDLCs). Methods Periodontal tissues collected from premolars for orthodontic treatment reasons were used to culture hPDLCs. Western blot analysis was performed to test the most optimal time and concentration of nicotine on the autophagy level of the hPDLCs. Transmission electron microscope and immunofluorescence observation were carried out to detect the form of autophagosomes and expression of autophagy related protein LC3 in hPDLCs under this optimal condition. Results Protein expression of LC3Ⅱ was up regulated with the 12 h nicotine stimulating. Besides that, the up regulation of the protein expression of LC3Ⅱ was concentration dependent and nicotine with a concentration of 1×10^-5mol·L^-1 was the most optimal condition. Transmis-sion electron microscope and immunofluorescence observations indicated that nicotine would activate the autophagy level of hPDLCs by increasing the number of autophagosomes and up regulating the expression of autophagy related protein LC3. Conclusion Nicotine could increase autophagy level of hPDLCs, thus affecting the occurrence and development of smoking related periodontitis.

Key words: human periodontal ligament cells, nicotine, autophagy

CLC Number:

R780.2

Yang Du, Shuai Yuan, Zhifei Zhou, Lizheng Wu, Lulu Wang, Xing’an Wu, Xiaojing Wang. A preliminary study on the autophagy level of human periodontal ligament cells regulated by nicotine[J]. West China Journal of Stomatology, 2017, 35(2): 198-202.

Figures/Tables 4

References 19

[1]	Wyganowska-Swiatkowska M, Nohawica MM.Effect of tobacco smoking on human gingival and periodontal fibro-blasts. A systematic review of literature[J]. Prz Lek, 2015, 72(3):158-160.
[2]	Chatzopoulos G.Smoking, smokeless tobacco, and alcohol consumption as contributing factors to periodontal disease[J]. Northwest Dent, 2016, 95(1):37-41.
[3]	Zhou Z, Li B, Dong Z, et al.Nicotine deteriorates the osteo-genic differentiation of periodontal ligament stem cells through α7 nicotinic acetylcholine receptor regulating Wnt pathway[J]. PLoS ONE, 2013, 8(12):e83102.
[4]	Wu L, Zhou Y, Zhou Z, et al.Nicotine induces the produc-tion of IL-1β and IL-8 via the α7 nAChR/NF-κB pathway in human periodontal ligament cells: an in vitro study[J]. Cell Physiol Biochem, 2014, 34(2):423-431.
[5]	Yang Z, Goronzy JJ, Weyand CM.Autophagy in autoimmune disease[J]. J Mol Med, 2015, 93(7):707-717.
[6]	Wu Y, Zhang Y, Wang L, et al.The role of autophagy in kidney inflammatory injury via the NF-κB route induced by LPS[J]. Int J Med Sci, 2015, 12(8):655-667.
[7]	Hung SY, Huang WP, Liou HC, et al.Autophagy protects neuron from Abeta-induced cytotoxicity[J]. Autophagy, 2009, 5(4):502-510.
[8]	Duerrschmidt N, Hagen A, Gaertner C, et al.Nicotine effects on human endothelial intercellular communication via α4β2 and α3β2 nicotinic acetylcholine receptor subtypes[J]. Naunyn Schmiedebergs Arch Pharmacol, 2012, 385(6):621-632.
[9]	Bullon P, Cordero MD, Quiles JL, et al.Autophagy in perio-dontitis patients and gingival fibroblasts: unraveling the link between chronic diseases and inflammation[J]. BMC Med, 2012, 10:122.
[10]	Suzuki H, Osawa T, Fujioka Y, et al.Structural biology of the core autophagy machinery[J]. Curr Opin Struct Biol, 2016, 43:10-17.
[11]	Tanida I, Ueno T, Kominami E.LC3 conjugation system in mammalian autophagy[J]. Int J Biochem Cell Biol, 2004, 36(12):2503-2518.
[12]	Hao Y, Lu Q, Yang G, et al.Lin28a protects against postin-farction myocardial remodeling and dysfunction through Sirt1 activation and autophagy enhancement[J]. Biochem Biophys Res Commun, 2016, 479(4):833-840.
[13]	Lu J, Sun D, Gao S, et al.Cyclovirobuxine D induces auto-phagy-associated cell death via the Akt/mTOR pathway in MCF-7 human breast cancer cells[J]. J Pharmacol Sci, 2014, 125(1):74-82.
[14]	Kim SY, Kang KL, Lee JC, et al.Nicotinic acetylcholine receptor α7 and β4 subunits contribute nicotine-induced apoptosis in periodontal ligament stem cells[J]. Mol Cells, 2012, 33(4):343-350.
[15]	Wang XJ, Liu YF, Wang QY, et al.Functional expression of alpha 7 nicotinic acetylcholine receptors in human perio-dontal ligament fibroblasts and rat periodontal tissues[J]. Cell Tissue Res, 2010, 340(2):347-355.
[16]	Liu YF, Wu LA, Wang J, et al.Micro-computerized tomo-graphy analysis of alveolar bone loss in ligature- and nicotine-induced experimental periodontitis in rats[J]. J Periodont Res, 2010, 45(6):714-719.
[17]	Song ZC, Zhou W, Shu R, et al.Hypoxia induces apoptosis and autophagic cell death in human periodontal ligament cells through HIF-1α pathway[J]. Cell Prolif, 2012, 45(3):239-248.
[18]	An Y, Liu W, Xue P, et al.Increased autophagy is required to protect periodontal ligament stem cells from apoptosis in inflammatory microenvironment[J]. J Clin Periodontol, 2016, 43(7):618-625.
[19]	Kim MS, Yun JW, Park JH, et al.Autophagy has a beneficial role in relieving cigarette smoke-induced apoptotic death in human gingival fibroblasts[J]. Int J Med Sci, 2016, 13(5):357-364.