Expression of autophagy-associated protein light chain 3B in human pulpitis and periapical periodontitis

doi:10.3969/j.issn.2095-4344.1177

Abstract

Abstract:

BACKGROUND: Role of autophagy in pulpitis, periapical periodontitis and alveolar bone destruction and the underling mechanisms have not been fully elucidated.
OBJECTIVE: To investigate the expression of autophagy-related protein light chain 3B in human pulpitis and periapical periodontitis, in order to explore the rule of autophagy.
METHODS: The healthy first premolar or surgically removed third premolar was selected from the patients undergoing orthodontics after their consent informs. The dental pulp cells were treated with 1 mg/L lipopolysaccharide for 0, 6, 12, 24 and 48 hours to induce inflammation. The expression of light chain 3B in normal and lipopolysaccharide-stimulated dental pulp cells was detected by western blot assay. The dotted accumulation of light chain 3B was observed by immunofluorescence. The expression level of light chain 3B in specimens of human chronic periapical periodontitis tissues was tested by real-time RT-PCR.
RESULTS AND CONCLUSION: The expression of light chain 3B in dental pulp cells stimulated by lipopolysaccharide for 6, 12, 24 and 48 hours was higher than that in normal dental pulp cells. The expression of light chain 3B increased with time, and peaked at 24 hours. Dotted accumulation of light chain 3B increased with lipopolysaccharide simulation time prolonged. The expression level of light chain 3B in specimens of human chronic periapical periodontitis tissues was higher than that in normal periapical tissues. These results indicate that the expression of light chain 3B is increased in human pulpitis and periapical periodontitis, and morphology also shows an increase of autophagy, suggesting that autophagy may be associated with the occurrence and development of pulpitis and periapical periodontitis.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Autophagy, Dental Pulp, Periapical Periodontitis, Tissue Engineering

CLC Number:

R496

Ma Jinbao, Wang Qian, Dong Ming, Bai Hua. Expression of autophagy-associated protein light chain 3B in human pulpitis and periapical periodontitis[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(15): 2364-2368.

Figures/Tables 3

References

[1] Coil J,Tam E,W aterfield JD.Proinflammatory cytokine p rofiles in pulp fibro-blasts stimulated with lipopolysaccharide and methylmer cap tan. J Endodon.2004; 30 (2): 88-91.

[2] Spilka CJ. Pathways of dental infections. J Oral Surg.1966;24(2): 111-124.

[3] Tagger M, Massler M.Periapical tissue reactions after pulp exposure in rat molars. Oral Surg Oral Med Oral Pathol. 1975, 39(2):304-317.

[4] Cecconi F, Levine B.The role of autophagy in mammalian development: cell makeover rather than cell death. Dev Cell.2008;15(3): 344-357．

[5] Levine B, Kroemer G. Autophagy in the Pathogenesis of Disease. Cell. 2008;132 (1): 27-42.

[6] Virgin HW, Levine B. Autophagy genes in immunity. Nat Immunol. 2009;10: 461-470.

[7] Gomes LC, Dikic I. Autophagy in Antimicrobial Immunity . Molecular Cell.2014;54: 224-233.

[8] Netea-Maier RT, Plantinga TS, van de Veerdonk FL, Smit JW, Netea MG. Modulation of inflammation by autophagy: consequences for human disease. Autophagy. 2016;12:245-260.

[9] Zhong Z, Sanchez-Lopez E, Karin M. Autophagy, NLRP3 inflammasome and auto-inflammatory/immune diseases. Clin Exp Rheumatol, 2016; 34:12–16.

[10] Pei F, Lin H, Liu H, et al. Dual role of autophagy in lipopolysaccharide -induced pre-odontoblastic cells. J Dent Res. 2015;94:175-182.

[11] Pei F,Wang HS,Chen Z,et al.Autophagy regulates odontoblast differentiation by suppressing NF-kappa B activation in an inflammatory environment. Cell Death Dis.2016;7: e2122.

[12] Wang HS,Pei F,Chen Z,et al. Increased apoptosis of inflamed odontoblasts is associated with CD47 loss. J Dent Res. 2016; 95(6): 697-703.

[13] Kuznetsov SA, Gelfand VI.18 kDa microtubule-associated protein: identification as a new light chain (LC-3) of microtubule-associated protein 1 (MAP-1). FEBS Lett.1987;212:145-148.

[14] Ladoire S, Chaba K, Martins I, et al. Immunohistochemical detection of cytoplasmic LC3 puncta in human cancer specimens. Autophagy. 2012;8:1175-1184.

[15] Tsukahara T, Matsuda Y, Haniu H.SF knockdown enhances apoptosis via downregulation of LC3B in human colon cancer cells. Biomed Res Int. 2013; 2013: 204973.

[16] Satyavarapu EM, Das R, Mandal C,et al. Autophagy-independent induction of LC3B through oxidative stress reveals its non-canonical role in anoikis of ovarian cancer cells. Cell Death Dis.2018; 9(10):934.

[17] Zhao H, Yang M, Zhao J, et al. High expression of LC3B is associated with progression and poor outcome in triple-negative breast cancer. Med Oncol.2013; 30(1):475.

[18] AlvarezVE, Kosec G, Sant AC, et al. Autophagy is involved innutritional stress response and differentiation in Trypanosom a cruzi. J BiolChem.2008; 283(6):3454-3464.

[19] Li L, Zhu YQ, Jiang L, et al. Increased autophagic activity in senescent human dental pulp cells. Int Endod J. 2012;45(12):1074-1079.

[20] Couve E,Schmachtenberg O.Autophagic activity and aging in human odontoblasts.J Dent Res. 2011; 90(4):523-528.

[21] Yang JW, Zhu LX, Yuan GH, et al. Autophagy appears during the development of the mouse lower first molar. Histochem Cell Biol. 2013;139(1):109-118.

[22] Bai H, Inoue J, Kawano T,et al. A transcriptional variant of the LC3A gene is involved in autophagy and frequently inactivated in human cancers. Oncogene.2012;31(40):4397-4408.

[23] Kapoor V,Paliwal D,Baskar Singh S,et al. Deregulation of Beclin 1 in patients with tobacco-related oral squamous cell carcinoma. Biochem Biophys Res Commun.2012;422(4):764-769.

[24] Bullon P, Cordero MD, Quiles JL, et al.Autophagy in periodontitis patients and gingival fibroblasts: unraveling the link between chronicdiseases and inflammation. BMC Med.2012;17(10):122-133.

[25] Kabeya Y, Mizushima N, Yamamoto A, et al. LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation. J Cell Sci.2004; 117: 2805-2812.

[26] Kakehashi S,Stanley H, Fitzgerald R. The effect of surgical exposures of dental pulps in germ-free and conventional laboratory rats. Oral Surg Oral Med Oral Pathol.1965;20(3):340-349.

[27] DeSelm CJ,Miller BC,Zou W,et al. Autophagy proteins regulate the secretory component of osteoclastic bone resorption. Dev Cell.2011; 21(5):966-974.

[28] Tchetina EV,Maslova KA,Krylov MY,et al. Association of bone loss with the upregulation of survival-related genes and concomitant downregulation of Mammalian target of rapamycin and osteoblast differentiation-related genes in the peripheral blood of late postmenopausal osteoporotic women . J Osteoporos.2015;2015: 802694.

[29] Morishita A,Kumabe S,Nakatsuka M,et al. A histological study of mineralised tissue formation around implants with 3D culture of HMS0014 cells in Cellmatrix Type I-A collagen gel scaffold in vitro . Okajimas Folia Anat Jpn.2014;91(3):57-71.

[30] Sun KT, Chen MY, Tu MG, et al.MicroRNA- 20a regulates autophagy related protein-ATG16L1 in hypoxia-induced osteoclast differentiation.Bone, 2015;73:145-153.

[31] Liu S, Zhu L, Zhang J, et al.Antiosteoclastogenic activity of isoliquiritigenin via inhibition of NF-κB-dependent autophagic pathway. Biochem Pharmacol.2016;106:82-93.

[32] Li Y, Guo T, Zhang Z, et al. Autophagy Modulates Cell Mineralization on Fluorapatite- Modified Scaffolds. J Dent Res. 2016;95(6):650-656.