HMBOX1 attenuates LPS‑induced periodontal ligament stem cell injury by inhibiting CXCL10 expression through the NF‑κB signaling pathway

Nie,Minyuan; Li,Heng; Liu,Puhe; Dang,Ping

doi:10.3892/etm.2022.11148

HMBOX1 attenuates LPS‑induced periodontal ligament stem cell injury by inhibiting CXCL10 expression through the NF‑κB signaling pathway

Authors:
- Minyuan Nie
- Heng Li
- Puhe Liu
- Ping Dang
View Affiliations

Affiliations: Department of Stomatology, PLA Strategic Support Force Characteristic Medical Center, Beijing 100101, P.R. China, Department of Paediatric Dentistry, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, Jiangsu 210008, P.R. China, Department of Stomatology, The First Dental Hospital, Wuhai, Inner Mongolia Autonomous Region 016000, P.R. China, Department of Stomatology, Amcare Women's and Children's Hospital, Beijing 100016, P.R. China
Published online on: January 17, 2022 https://doi.org/10.3892/etm.2022.11148
Article Number: 224
Copyright: © Nie et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Homeobox containing 1 (HMBOX1) is a member of the homeobox transcription factor family that has been reported to serve an important role in numerous biological processes. The present study aimed to determine the role of HMBOX1 in the pathogenesis of periodontitis. Human periodontal ligament stem cells (hPDLSCs) were treated with liposaccharide (LPS) and transfected with a HMBOX1 overexpression (Ov‑HMBOX1) plasmid or small interfering (si)‑C‑X‑C motif chemokine ligand 10 (CXCL10) plasmids. The effects of Ov‑HMBOX1 on cell proliferation, inflammation and apoptosis were subsequently investigated using Cell Counting Kit‑8, ELISA for analysis of IL‑6, TNF‑α and IL‑1β levels, TUNEL and western blotting assays for analysis of Bcl‑2, Bax, cleaved caspase‑3 and caspase‑3 levels, respectively. Furthermore, the potential effects of HMBOX1 on the mRNA and protein levels of CXCL10 and the NF‑κB signaling pathway were investigated by using reverse transcription‑quantitative PCR and western blotting. Finally, the physiological processes of lipopolysaccharide (LPS)‑induced hPDLSCs overexpressing HMBOX1 were assessed following treatment with phorbol 12‑myristate 13‑acetate (PMA), a NF‑κB agonist. The results revealed that Ov‑HMBOX1 transfection promoted proliferation whilst alleviating inflammation and apoptosis in LPS‑induced hPDLSCs. Ov‑HMBOX1 reduced the expression of CXCL10 by suppressing the NF‑κB signaling pathway. PMA treatment inhibited the proliferation of LPS‑induced hPDLSCs transfected with Ov‑HMBOX1, which was reversed by transfection with si‑CXCL10. In conclusion, results of the present study provided evidence that HMBOX1 can attenuate LPS‑induced hPDLSC injury by downregulating CXCL10 expression via the NF‑κB signaling pathway, which may provide a novel insight into the development of potentially novel treatment strategies for periodontitis.

View Figures

View References

1	Krishna R and De Stefano JA: Ultrasonic vs. hand instrumentation in periodontal therapy: Clinical outcomes. Periodontol 2000. 71:113–127. 2016.PubMed/NCBI View Article : Google Scholar
2	Van Dyke TE: The management of inflammation in periodontal disease. J Periodontol. 79 (Suppl 8):S1601–S1608. 2008.PubMed/NCBI View Article : Google Scholar
3	Papapanou PN, Sanz M, Buduneli N, Dietrich T, Feres M, Fine DH, Flemmig TF, Garcia R, Giannobile WV, Graziani F, et al: Periodontitis: Consensus report of workgroup 2 of the 2017 World Workshop on the classification of periodontal and Peri-implant diseases and conditions. J Periodontol. 89 (Suppl 1):S173–S182. 2018.PubMed/NCBI View Article : Google Scholar
4	Slots J: Periodontitis: Facts, fallacies and the future. Periodontol 2000. 75:7–23. 2017.PubMed/NCBI View Article : Google Scholar
5	Borgnakke WS, Ylostalo PV, Taylor GW and Genco RJ: Effect of periodontal disease on diabetes: Systematic review of epidemiologic observational evidence. J Periodontol. 40 (Suppl 14):S135–S152. 2013.PubMed/NCBI View Article : Google Scholar
6	Manresa C, Sanz-Miralles EC, Twigg J and Bravo M: Supportive periodontal therapy (SPT) for maintaining the dentition in adults treated for periodontitis. Cochrane Database Syst Rev. 1(CD009376)2018.PubMed/NCBI View Article : Google Scholar
7	Chen S, Saiyin H, Zeng X, Xi J, Liu X, Li X and Yu L: Isolation and functional analysis of human HMBOX1, a homeobox containing protein with transcriptional repressor activity. Cytogenet Genome Res. 114:131–136. 2006.PubMed/NCBI View Article : Google Scholar
8	Diao N, Li Y, Yang J, Jin C, Meng X, Jiao W, Feng J, Liu Z and Lu N: High expression of HMBOX1 contributes to poor prognosis of gastric cancer by promoting cell proliferation and migration. Biomed Pharmacother. 115(108867)2019.PubMed/NCBI View Article : Google Scholar
9	Yuan HX, Feng XE, Liu EL, Ge R, Zhang YL, Xiao BG and Li QS: 5,2'-dibromo-2,4',5'-trihydroxydiphenylmethanone attenuates LPS-induced inflammation and ROS production in EA.hy926 cells via HMBOX1 induction. J Cell Mol Med. 23:453–463. 2019.PubMed/NCBI View Article : Google Scholar
10	Ma H, Su L, He X and Miao J: Loss of HMBOX1 promotes LPS-induced apoptosis and inhibits LPS-induced autophagy of vascular endothelial cells in mouse. Apoptosis. 24:946–957. 2019.PubMed/NCBI View Article : Google Scholar
11	Guzeldemir-Akcakanat E, Alkan B, Sunnetci-Akkoyunlu D, Gurel B, Balta VM, Kan B, Akgun E, Yilmaz EB, Baykal AT, Cine N, et al: Molecular signatures of chronic periodontitis in gingiva: A genomic and proteomic analysis. J Periodontol. 90:663–673. 2019.PubMed/NCBI View Article : Google Scholar
12	Zhao H, Han Q, Lu N, Xu D, Tian Z and Zhang J: HMBOX1 in hepatocytes attenuates LPS/D-GalN-induced liver injury by inhibiting macrophage infiltration and activation. Mol Immunol. 101:303–311. 2018.PubMed/NCBI View Article : Google Scholar
13	Zhang F, Mears JR, Shakib L, Beynor JI, Shanaj S, Korsunsky I and Nathan A: Accelerating Medicines Partnership Rheumatoid Arthritis and Systemic Lupus Erythematosus (AMP RA/SLE) Consortium. Donlin LT and Raychaudhuri S: IFN-γ and TNF-α drive a CXCL10⁺ CCL2⁺ macrophage phenotype expanded in severe COVID-19 and other diseases with tissue inflammation. bioRxiv: Aug 5, 2020 (Epub ahead of print). doi: 10.1101/2020.08.05.238360.
14	Luster AD, Unkeless JC and Ravetch JV: Gamma-interferon transcriptionally regulates an early-response gene containing homology to platelet proteins. Nature. 315:672–676. 1985.PubMed/NCBI View Article : Google Scholar
15	Groom JR and Luster AD: CXCR3 ligands: Redundant, collaborative and antagonistic functions. Immunol Cell Biol. 89:207–215. 2011.PubMed/NCBI View Article : Google Scholar
16	Gao J, Wu L, Wang S and Chen X: Role of chemokine (C-X-C Motif) ligand 10 (CXCl10) in renal diseases. Mediators Inflamm. 2020(6194864)2020.PubMed/NCBI View Article : Google Scholar
17	Aldahlawi S, Youssef AR and Shahabuddin S: Evaluation of chemokine CXCL10 in human gingival crevicular fluid, saliva, and serum as periodontitis biomarker. J Inflamm Res. 11:389–396. 2018.PubMed/NCBI View Article : Google Scholar
18	You N, Li J, Huang X, Wu K, Tang Y, Wang L, Li H, Mi N and Zheng L: COMMD7 activates CXCl10 production by regulating NF-κB and the production of reactive oxygen species. Mol Med Rep. 17:6784–6788. 2018.PubMed/NCBI View Article : Google Scholar
19	Chen H, Lin W, Lin P, Zheng M, Lai Y, Chen M, Zhang Y, Chen J, Lin X, Lin L, et al: IL-10 produces a dual effect on OGD-induced neuronal apoptosis of cultured cortical neurons via the NF-κB pathway. Aging (Albany NY). 11:10796–10813. 2019.PubMed/NCBI View Article : Google Scholar
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
21	Zhou W, Su L, Duan X, Chen X, Hays A, Upadhyayula S, Shivde J, Wang H, Li Y, Huang D and Liang S: MicroRNA-21 down-regulates inflammation and inhibits periodontitis. Mol Immunol. 101:608–614. 2018.PubMed/NCBI View Article : Google Scholar
22	Holland PW, Booth HA and Bruford EA: Classification and nomenclature of all human homeobox genes. BMC Biol. 5(47)2007.PubMed/NCBI View Article : Google Scholar
23	Chen S, Li Y, Zhi S, Ding Z, Wang W, Peng Y, Huang Y, Zheng R, Yu H, Wang J, et al: WTAP promotes osteosarcoma tumorigenesis by repressing HMBOX1 expression in an m⁶A-dependent manner. Cell Death Dis. 11(659)2020.PubMed/NCBI View Article : Google Scholar
24	Li J, Zhao Y and Wang J: Extracellular vesicle-associated microRNA-221-3p secreted by drug-resistant lung cancer cells targets HMBOX1 to promote the progression of lung cancer. Cancer Gene Ther. 28:679–692. 2021.PubMed/NCBI View Article : Google Scholar
25	Zhao H, Jia H, Han Q and Zhang J: Homeobox containing 1 inhibits liver cancer progression by promoting autophagy as well as inhibiting stemness and immune escape. Oncol Rep. 40:1657–1665. 2018.PubMed/NCBI View Article : Google Scholar
26	Tsai CF, Chen JH and Yeh WL: Pulmonary fibroblasts-secreted CXCL10 polarizes alveolar macrophages under pro-inflammatory stimuli. Toxicol Appl Pharmacol. 380(114698)2019.PubMed/NCBI View Article : Google Scholar
27	McGrath-Morrow SA, Lee S, Gibbs K, Lopez A, Collaco JM, Neptune E, Soloski MJ, Scott A and D'Alessio F: Immune response to intrapharyngeal LPS in neonatal and juvenile mice. Am J Respir Cell Mol Biol. 52:323–331. 2015.PubMed/NCBI View Article : Google Scholar
28	Chalin A, Lefevre B, Devisme C, Pronier C, Carrière V, Thibault V, Amiot L and Samson M: Serum CXCL10, CXCL11, CXCL12, and CXCL14 chemokine patterns in patients with acute liver injury. Cytokine. 111:500–504. 2018.PubMed/NCBI View Article : Google Scholar
29	Zhai Y, Shen XD, Gao F, Zhao A, Freitas MC, Lassman C, Luster AD, Busuttil RW and Kupiec-Weglinski JW: CXCL10 regulates liver innate immune response against ischemia and reperfusion injury. Hepatology. 47:207–214. 2008.PubMed/NCBI View Article : Google Scholar
30	Shimada Y, Tabeta K, Sugita N and Yoshie H: Profiling biomarkers in gingival crevicular fluid using multiplex bead immunoassay. Arch Oral Biol. 58:724–730. 2013.PubMed/NCBI View Article : Google Scholar
31	Dong M, Yu X, Chen W, Guo Z, Sui L, Xu Y, Shang Y, Niu W and Kong Y: Osteopontin promotes bone destruction in periapical periodontitis by activating the NF-κB pathway. Cell Physiol Biochem. 49:884–898. 2018.PubMed/NCBI View Article : Google Scholar
32	Hiyari S, Wong RL, Yaghsezian A, Naghibi A, Tetradis S, Camargo PM and Pirih FQ: Ligature-induced peri-implantitis and periodontitis in mice. J Clin Periodontol. 45:89–99. 2018.PubMed/NCBI View Article : Google Scholar
33	Huang Y, Zeng J, Chen G, Xie X, Guo W and Tian W: Periodontitis contributes to adipose tissue inflammation through the NF-B, JNK and ERK pathways to promote insulin resistance in a rat model. Microbes Infect. 18:804–812. 2016.PubMed/NCBI View Article : Google Scholar
34	Jin WJ, Kim B, Kim D, Park Choo HY, Kim HH, Ha H and Lee ZH: NF-κB signaling regulates cell-autonomous regulation of CXCL10 in breast cancer 4T1 cells. Exp Mol Med. 49(e295)2017.PubMed/NCBI View Article : Google Scholar