XBP1s调控炎症诱导的成牙本质细胞自噬

doi:10.13701/j.cnki.kqyxyj.2021.05.009

摘要/Abstract

摘要： 目的:探讨转录因子剪接型X-盒结合蛋白1 (spliced X-box binding protein 1,XBP1s)对炎症诱导的成牙本质细胞自噬的作用。方法:免疫组化检测XBP1和自噬指标LC3在健康和炎症牙髓组织中的表达。利用脂多糖(lipopolysaccharide , LPS)刺激小鼠牙乳头细胞系(mDPC6T)建立体外炎症模型,检测XBP1和自噬指标LC3、ATG5的表达,再分别利用抑制剂KIRA6,和活性氧(Reactive oxygen species,ROS)清除剂NAC处理细胞。通过免疫荧光和蛋白免疫印迹法检测XBP1s、LC3和ATG5等指标的表达。结果:XBP1s和LC3在炎症牙髓的成牙本质细胞层中表达升高。LPS可诱导XBP1s在mDPC6T细胞中表达,与LC3、ATG5等自噬指标的表达趋势相似。抑制XBP1s可降低LC3、ATG5等自噬指标的表达。结论:XBP1s可调控炎症诱导的成牙本质细胞自噬。

关键词: XBP1s, 自噬, 成牙本质细胞, 炎症

Abstract: Objective: To examine the function of XBP1s in inflammation-induced autophagy in odontoblasts. Methods: Immunohistochemistry was used to examine the expression of XBP1 and autophagy markers (LC3 and ATG5) in human dental pulp samples. mDPC6T cells were stimulated by lipopolysaccharide (LPS) to establish an in vitro inflammatory model. The cells were further treated with the inhibitor KIRA6 and the reactive oxygen species (ROS) scavenging agent NAC, respectively. Immunofluorescence and western blotting were used to detect the expression of XBP1s and autophagy-related markers (LC3 and ATG5). Results: The expressions of XBP1, autophagy markers, and apoptosis markers were upregulated in odontoblast layers of human pulpitis tissues. LPS could induce the expression of spliced XBP1 in odontoblasts, and the expression of spliced XBP1 correlated with the expression of autophagy regulators under severe LPS stimulation. Moreover, the suppression of spliced XBP1 could reduce autophagy and apoptosis in inflamed odontoblasts. In addition, a reactive oxygen species (ROS) scavenger decreased the level of spliced XBP1 in inflamed mDPC6T cells. Conclusion: Spliced XBP1 can regulate autophagy in inflamed odontoblasts

Key words: XBP1s, autophagy, odontoblasts, inflammation

王艳杰, 王海胜, 郭威廷, 张露. XBP1s调控炎症诱导的成牙本质细胞自噬[J]. 口腔医学研究, 2021, 37(5): 418-424.

WANG Yanjie, WANG Haisheng, GUO Weiting, ZHANG Lu. Spliced XBP1 Regulates Autophagy in Inflamed Odontoblasts[J]. Journal of Oral Science Research, 2021, 37(5): 418-424.

参考文献

[1] Farges JC, Alliot-Licht B, Baudouin C, et al. Odontoblast control of dental pulp inflammation triggered by cariogenic bacteria [J]. Front Physiol, 2013, 4:326.
[2] Couve E, Schmachtenberg O. Autophagic activity and aging in human odontoblasts [J]. J Dent Res, 2011, 90(4):523-528.
[3] Wang X, Wu TT, Jiang L, et al. Deferoxamine-induced migration and odontoblast differentiation via ROS-dependent autophagy in dental pulp stem cells [J]. Cell Physiol Biochem, 2017, 43(6):2535-2547.
[4] Doherty J, Baehrecke EH. Life, death and autophagy [J]. Nat Cell Biol, 2018, 20(10):1110-1117.
[5] Kuma A, Komatsu M, Mizushima N. Autophagy-monitoring and autophagy-deficient mice [J]. Autophagy, 2017, 13(10):1619-1628.
[6] 黄佩,乔玮玮,孟柳燕.自噬通过GATA4调控人牙髓细胞衰老反应[J].口腔医学研究,2020,36(12):1142-1147.
[7] Wang HS, Pei F, Chen Z, et al. Increased apoptosis of inflamed odontoblasts is associated with CD47 loss [J]. J Dent Res, 2016, 95(6):697-703.
[8] Li Y, Wang H, Pei F, et al. FoxO3a regulates inflammation-induced autophagy in odontoblasts [J]. J Endod, 2018, 44(5):786-791.
[9] Guo T, Cao G, Liu B, et al. Cbfalpha1 hinders autophagy by DSPP upregulation in odontoblast differentiation [J]. Int J Biochem Cell Biol, 2019, 115:105578.
[10] Huang S, Xing Y, Liu Y. Emerging roles for the ER stress sensor IRE1alpha in metabolic regulation and disease [J]. J Biol Chem, 2019, 294(49):18726-18741.
[11] Cho HK, Kim SY, Kyaw YY, et al. HBx induces the proliferation of hepatocellular carcinoma cells via AP1 over-expressed as a result of ER stress [J]. Biochem J, 2015, 466(1):115-121.
[12] Ochoa CD, Wu RF, Terada LS. ROS signaling and ER stress in cardiovascular disease [J]. Mol Aspects Med, 2018, 63:18-29.
[13] Hetz C, Saxena S. ER stress and the unfolded protein response in neurodegeneration [J]. Nat Rev Neurol, 2017, 13(8):477-491.
[14] van Schadewijk A, Van'T WE, Stolk J, et al. A quantitative method for detection of spliced X-box binding protein-1 (XBP1) mRNA as a measure of endoplasmic reticulum (ER) stress [J]. Cell Stress Chaperones, 2012, 17(2):275-279.
[15] Lee AH, Iwakoshi NN, Glimcher LH. XBP-1 regulates a subset of endoplasmic reticulum resident chaperone genes in the unfolded protein response [J]. Mol Cell Biol, 2003, 23(21):7448-7459.
[16] Wu S, Du R, Gao C, et al. The role of XBP1s in the metastasis and prognosis of hepatocellular carcinoma [J]. Biochem Biophys Res Commun, 2018, 500(3):530-537.
[17] Xu W, Wang C, Hua J. X-box binding protein 1 (XBP1) function in diseases [J]. Cell Biol Int, 2021, 45(4):731-739.
[18] Zhang X, Hu P, Ding SY, et al. Induction of autophagy-dependent apoptosis in cancer cells through activation of ER stress: an uncovered anti-cancer mechanism by anti-alcoholism drug disulfiram [J]. Am J Cancer Res, 2019, 9(6):1266-1281.
[19] Tian PG, Jiang ZX, Li JH, et al. Spliced XBP1 promotes macrophage survival and autophagy by interacting with Beclin-1 [J]. Biochem Biophys Res Commun, 2015, 463(4):518-523.
[20] Pezelj-Ribaric S, Anic I, Brekalo I, et al. Detection of tumor necrosis factor alpha in normal and inflamed human dental pulps [J]. Arch Med Res, 2002, 33(5):482-484.
[21] Lin H, Liu H, Sun Q, et al. Establishment and characterization of a tamoxifen-mediated reversible immortalized mouse dental papilla cell line [J]. In Vitro Cell Dev Biol Anim, 2013, 49(2):114-121.
[22] 陈杰,徐华兴,程抒华,等.细胞自噬在牙髓损伤修复过程中的作用[J].口腔医学研究,2019,35(6):531-536.
[23] Iwakoshi NN, Lee AH, Glimcher LH. The X-box binding protein-1 transcription factor is required for plasma cell differentiation and the unfolded protein response [J]. Immunol Rev, 2003, 194:29-38.
[24] Larabi A, Barnich N, Nguyen H. New insights into the interplay between autophagy, gut microbiota and inflammatory responses in IBD [J]. Autophagy, 2020, 16(1):38-51.
[25] Chen J, Zhang ZQ, Song J, et al. 18beta-Glycyrrhetinic-acid-mediated unfolded protein response induces autophagy and apoptosis in hepatocellular carcinoma [J]. Sci Rep, 2018, 8(1):9365.
[26] Kishino A, Hayashi K, Hidai C, et al. XBP1-FoxO1 interaction regulates ER stress-induced autophagy in auditory cells [J]. Sci Rep, 2017, 7(1):4442.
[27] Tian PG, Jiang ZX, Li JH, et al. Spliced XBP1 promotes macrophage survival and autophagy by interacting with Beclin-1 [J]. Biochem Biophys Res Commun, 2015, 463(4):518-523.
[28] Pei F, Lin H, Liu H, et al. Dual role of autophagy in lipopolysaccharide-induced preodontoblastic cells [J]. J Dent Res, 2015, 94(1):175-182.
[29] Jiang D, Niwa M, Koong AC. Targeting the IRE1alpha-XBP1 branch of the unfolded protein response in human diseases [J]. Semin Cancer Biol, 2015, 33:48-56.
[30] Yang F, Li Y, Duan H, et al. Activation of mitophagy in inflamed odontoblasts [J]. Oral Dis, 2019, 25(6):1581-1588.
[31] Sharma M, Sharma KB, Chauhan S, et al. Diphenyleneiodonium enhances oxidative stress and inhibits Japanese encephalitis virus induced autophagy and ER stress pathways [J]. Biochem Biophys Res Commun, 2018, 502(2):232-237.
[32] Chen HY, Ren XY, Wang WH, et al. Upregulated ROS production induced by the proteasome inhibitor MG-132 on XBP1 gene expression and cell apoptosis in Tca-8113 cells [J]. Biomed Pharmacother, 2014, 68(6):709-713.