Expression of autophagy-related protein Beclin-1 and microtubule-associated protein 2 light chain 3 in periodontal ligament cells in orthodontic tooth pressure areas

doi:10.7518/hxkq.2019.02.008

Abstract

Abstract:

Objective To investigate the expression of autophagy-related protein Beclin-1 and microtubule-associated protein 2 light chain 3 (LC3Ⅱ) in periodontal ligament cells in orthodontic tooth pressure areas. Methods Sixty male SD rats were randomly divided into a blank control group and nine experimental groups. In the experimental groups, 0.392 N orthodontic force was used to move the first right upper molars for 15 min, 30 min, 1 h, 2 h, 4 h, 12 h, 1 d, 3 d, or 7 d. The blank control group did not receive any treatment. The rats were euthanized. Changes in the morphology of the periodontal membrane in the pressure areas were observed through hematoxylin and eosin (HE) staining. The expression levels of Beclin-1 and LC3Ⅱ were detected by immunohistochemical staining, and tartrate-resistant acid phosphatase (TRAP) staining was performed for the counting of osteoclasts. Results The HE stains showed that the hyalinization of the periodontal ligament appeared in the pressure areas after 1 day of exertion and was gradually aggravated. The immunohistochemical stains showed that the expression levels of Beclin-1 and LC3Ⅱ in the experimental groups gradually increased, peaked after 1 h, and then gradually decreased. The expression levels peaked again after 1 d, then decreased to baseline levels at 7 d of exertion. Beclin-1 and LC3Ⅱ were expressed in the osteoclasts. The TRAP stains indicated that the number of osteoclasts started to increase after 1 day. Conclusion Autophagy may participate in the process of periodontal ligament reconstruction in orthodontic tooth pressure areas by mediating the hyalini-zation of periodontal ligament and affecting the biological effects of osteoclasts.

Key words: orthodontic tooth movement, periodontal ligament, pressure, autophagy

CLC Number:

R783.5

Jialing Lü,Jie Xu,Jin Zeng,Haixia Dang,Jinghong Yu,Xian Zhao,Xiaomei Xu. Expression of autophagy-related protein Beclin-1 and microtubule-associated protein 2 light chain 3 in periodontal ligament cells in orthodontic tooth pressure areas[J]. West China Journal of Stomatology, 2019, 37(2): 168-173.

Figures/Tables 8

Fig.1

Fig.2

Fig.3

Fig.4

Fig.5

Fig.6

Fig.7

Fig.8

References 13

[1]	Dutra EH, Nanda R, Yadav S . Bone response of loaded periodontal ligament[J]. Curr Osteoporos Rep, 2016,14(6):280-283. doi: 10.1007/s11914-016-0328-x URL
[2]	King JS, Veltman DM, Insall RH . The induction of auto-phagy by mechanical stress[J]. Autophagy, 2011,7(12):1490-1499. doi: 10.4161/auto.7.12.17924 URL
[3]	Ma KG, Shao ZW, Yang SH , et al. Autophagy is activated in compression-induced cell degeneration and is mediated by reactive oxygen species in nucleus pulposus cells exposed to compression[J]. Osteoarthritis Cartilage, 2013,21(12):2030-2038. doi: 10.1016/j.joca.2013.10.002 URL
[4]	Baskaran R, Poornima P, Priya LB , et al. Neferine prevents autophagy induced by hypoxia through activation of Akt/mTOR pathway and Nrf2 in muscle cells[J]. Biomed Phar-macother, 2016,83:1407-1413. doi: 10.1016/j.biopha.2016.08.063 URL
[5]	Huang ZJ, Fang WL, Liu WH , et al. Aspirin induces Beclin-1-dependent autophagy of human hepatocellular carcinoma cell[J]. Eur J Pharmacol, 2018,823:58-64. doi: 10.1016/j.ejphar.2018.01.031 URL
[6]	Adiseshaiah PP, Skoczen SL, Rodriguez JC , et al. Autophagy monitoring assay Ⅱ: imaging autophagy induction in LLC-PK1 cells using GFP-LC3 protein fusion construct[J]. Me-thods Mol Biol, 2018,1682:211-219. doi: 10.1007/978-1-4939-7352-1 URL
[7]	Mariño G, Niso-Santano M, Baehrecke EH , et al. Self-con-sumption: the interplay of autophagy and apoptosis[J]. Nat Rev Mol Cell Biol, 2014,15(2):81-94. doi: 10.1038/nrm3735 URL
[8]	Chen X, Wu JX, You XJ , et al. Cold ischemia-induced auto-phagy in rat lung tissue[J]. Mol Med Rep, 2015,11(4):2513-2519. doi: 10.3892/mmr.2014.2999 URL
[9]	Rody WJ, King GJ, Gu GM . Osteoclast recruitment to sites of compression in orthodontic tooth movement[J]. Am J Orthod Dentofacial Orthop, 2001,120(5):477-489. doi: 10.1067/mod.2001.118623 URL
[10]	Zhao Y, Chen G, Zhang W , et al. Autophagy regulates hypoxia-induced osteoclastogenesis through the HIF-1α/BNIP3 signaling pathway[J]. J Cell Physiol, 2012,227(2):639-648. doi: 10.1002/jcp.22768 URL
[11]	Shi J, Wang L, Zhang HY , et al. Glucocorticoids: dose-related effects on osteoclast formation and function via reactive oxygen species and autophagy[J]. Bone, 2015,79:222-232. doi: 10.1016/j.bone.2015.06.014 URL
[12]	Wu DJ, Adamopoulos IE . Autophagy and autoimmunity[J]. Clin Immunol, 2017,176:55-62. doi: 10.1016/j.clim.2017.01.007 URL
[13]	Wijekoon S, Bwalya EC, Fang J , et al. Chronological dif-ferential effects of pro-inflammatory cytokines on RANKL-induced osteoclast differentiation of canine bone marrow-derived macrophages[J]. J Vet Med Sci, 2017,79(12):2030-2035. doi: 10.1292/jvms.17-0393 URL

[1]	Tang Xiaoxue, Zhou Zheng, Li Qiqi, Jiang Dandan. Effects of sitagliptin activation of the stromal cell-derived factor-1/CXC chemokine receptor 4 signaling pathway on the proliferation, apoptosis, inflammation, and osteogenic differentiation of human periodontal ligament stem cells induced by lipopolysaccharide [J]. West China Journal of Stomatology, 2024, 42(1): 37-45.
[2]	Tang Meng, Cui Zhan-qin, Wang Yangyang, Chen Zengguo, Li Wenjing, Zhang Cuiping. Effects of low-level laser on the expression of interleukin-6, tumor necrosis factor‑α, osteoprotegerin, and receptor activator of nuclear factor-κB ligand in human periodontal ligament cells [J]. West China Journal of Stomatology, 2023, 41(5): 521-532.
[3]	Wan Xiaofang, He Haiyan, Jialing Lü, Wu Yujie, Zhong Guannan, Xu Xiaomei. Hippo-YAP signaling pathway regulates autophagy of human periodontal ligament cells under cyclic tensile stress [J]. West China Journal of Stomatology, 2023, 41(3): 260-268.
[4]	Lu Ting, Zhu Jiahao, Yang Shihe, Shen Zhe, Zhong Liangjun. Effects of Foxp3 gene silencing on the expression of inflammatory cytokines and the proliferation and migration of human periodontal ligament fibroblasts in an inflammatory environment [J]. West China Journal of Stomatology, 2023, 41(3): 269-275.
[5]	Li Shenghong, Peng Shiyuan, Luo Xiaoling, Wang Yipei, Xu Xiaomei. Effect of naringenin on the anti-inflammatory, vascularization, and osteogenesis differentiation of human periodontal ligament stem cells via the stromal cell-derived factor 1/C-X-C motif chemokine receptor 4 signaling axis stimulated by lipopolysaccharide [J]. West China Journal of Stomatology, 2023, 41(2): 175-184.
[6]	Fan Zhibo, Jin Ke, Li Shenghong, Xu Jie, Xu Xiaomei. Regulation of reactive oxygen species on the mitophagy of human periodontal ligament cells through the PINK1/Parkin pathway under starvation [J]. West China Journal of Stomatology, 2022, 40(6): 645-653.
[7]	Wu Jie, Cui Zhanqin, Han Yu, Li Wenjing. Aging effect of osteoprotegerin and receptor activator of nuclear factor-κB ligand expression in human periodontal ligament cells under continuous static pressure [J]. West China Journal of Stomatology, 2022, 40(6): 654-661.
[8]	Zhao Zhihe, Jin Zuolin, Bai Yuxing, Fang Bing, Bai Ding, Li Weiran, He Hong, Hu Min, Liu Yuehua, Chen Lili, Song Jinlin, Cao Yang, Li Yu, Shu Rui. Core scientific issues of orthodontic tooth movement: position objective, efficiency, and accuracy [J]. West China Journal of Stomatology, 2022, 40(4): 371-376.
[9]	Dai Zhenning, Zheng Weihan, Li Shiyu. Receptor activator of nuclear factor-κB ligand and tumor necrosis factor-α promotes osteoclast differentiation through the exosomes of inflammatory periodontal ligament stem cells [J]. West China Journal of Stomatology, 2022, 40(4): 377-385.
[10]	Pang Ming, Wei Hongxia, Chen Xi. Long non-coding RNA potassium voltage-gated channel subfamily Q member 1 overlapping transcript 1 regulates the proliferation and osteogenic differentiation of human periodontal ligament stem cells by targeting miR-24-3p [J]. West China Journal of Stomatology, 2021, 39(5): 547-554.
[11]	Jin Xiaolan, Zhang Yanan, Sun Chengrui, Zou Zhaohui. Protective effect of low-level laser irradiation on lipopolysaccharide-mediated inflammatory injury of human periodontal ligament fibroblasts [J]. West China Journal of Stomatology, 2021, 39(3): 260-266.
[12]	Wang Luyao, Wang Bo, Cong Beibei, Cui Tao, Du Yuqing, Song Yu. Effect of different orthodontic forces on the expression of T helper cell 17 cell-related cytokines in the pressure side of periodontal tissue in rats [J]. West China Journal of Stomatology, 2021, 39(2): 143-152.
[13]	Wang Lin, Wang Xi, Ji Nan, Li Haimei, Cai Shixin. Mechanisms of the mechanically activated ion channel Piezo1 protein in mediating osteogenic differentiation of periodontal ligament stem cells via the Notch signaling pathway [J]. West China Journal of Stomatology, 2020, 38(6): 628-636.
[14]	Liu Xinchen, Lu Jinjin, Chen Yumeng, Qiu Ying, Zheng Mengdan, Wang Zilin, Li Xiangwei. Roles of autophagy onself-renewal and differentiation of mesenchymal stem cells [J]. West China Journal of Stomatology, 2020, 38(6): 704-707.
[15]	Wen Jiahui, Wu Yanmin, Chen Lili. Functions of non-coding RNAs in the osteogenic differentiation of human periodontal ligament-derived cells [J]. West China Journal of Stomatology, 2020, 38(3): 330-337.