Nie Minyuan, Li Heng, Liu Puhe, Dang Ping
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.
Exp Ther Med. 2022 Mar;23(3):224. doi: 10.3892/etm.2022.11148. Epub 2022 Jan 17.
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.
含同源框1(HMBOX1)是同源框转录因子家族的成员,据报道在众多生物学过程中发挥重要作用。本研究旨在确定HMBOX1在牙周炎发病机制中的作用。用人牙周膜干细胞(hPDLSCs)进行脂多糖(LPS)处理,并转染HMBOX1过表达(Ov-HMBOX1)质粒或小干扰(si)-C-X-C基序趋化因子配体10(CXCL10)质粒。随后使用细胞计数试剂盒-8、用于分析IL-6、TNF-α和IL-1β水平的ELISA、分别用于分析Bcl-2、Bax、裂解的caspase-3和caspase-3水平的TUNEL和蛋白质印迹分析,研究Ov-HMBOX1对细胞增殖、炎症和凋亡的影响。此外,通过逆转录定量PCR和蛋白质印迹研究HMBOX1对CXCL10的mRNA和蛋白质水平以及NF-κB信号通路的潜在影响。最后,在用佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)(一种NF-κB激动剂)处理后,评估过表达HMBOX1的脂多糖(LPS)诱导的hPDLSCs的生理过程。结果显示,Ov-HMBOX1转染促进了LPS诱导的hPDLSCs的增殖,同时减轻了炎症和凋亡。Ov-HMBOX1通过抑制NF-κB信号通路降低了CXCL10的表达。PMA处理抑制了用Ov-HMBOX1转染的LPS诱导的hPDLSCs的增殖,而用si-CXCL10转染可逆转这种抑制。总之,本研究结果提供了证据,表明HMBOX1可通过NF-κB信号通路下调CXCL10表达来减轻LPS诱导的hPDLSC损伤,这可能为牙周炎潜在新治疗策略的开发提供新的见解。