应激颗粒形成对人牙周膜细胞凋亡及对LPS刺激下炎症相关因子表达及ROS生成的影响

doi:10.12439/kqhm.1005-4979.2025.03.003

摘要/Abstract

摘要：

目的：探讨应激状态下人牙周膜细胞（human periodontal ligament cells，hPDLCs）中应激颗粒（stress granule，SG）的形成及其对细胞凋亡、脂多糖（lipopolysaccharide，LPS）诱导下炎症相关因子表达和活性氧（reactive oxygen species，ROS）生成的影响。方法：采用亚砷酸钠（sodium arsenite，SA）诱导 hPDLCs 形成 SG，通过免疫荧光染色对 SG进行量化分析；通过活死细胞染色、实时定量聚合酶链反应（real-time quantitative polymerase chain reaction，RT-qPCR）和免疫荧光检测 hPDLCs 中半胱氨酰天冬氨酸特异性蛋白酶 3（cysteinyl aspartate-specific proteinase 3，Caspase-3）的定位及表达，评估 SG 对细胞凋亡的影响；采用 RT-qPCR 检测 LPS 刺激下炎症因子白细胞介素1β（interleukin 1β，IL-1β）和白细胞介素 6 （interleukin 6，IL-6）的 mRNA 表达变化；应用 ROS 检测试剂盒检测 SG 对 LPS 诱导下 hPDLCs 中的 ROS 水平影响。结果：SA 成功诱导 hPDLCs 形成 SG；免疫荧光结果显示 Caspase-3 定位于 SG，且 SG 的形成显著抑制了 hPDLCs 凋亡；RT-qPCR 结果显示，SG 可下调 LPS 刺激下 hPDLCs 相关炎症因子的 mRNA 相对表达；ROS 检测结果显示 SG 能抑制 LPS 刺激下 hPDLCs 的 ROS 生成。结论：SG 可保护 hPDLCs 免受应激诱导的细胞凋亡，并显著抑制LPS 触发的炎症因子的表达及 ROS 生成。

关键词:

应激颗粒, 凋亡, 牙周炎, 活性氧

Abstract:

Objective: To investigate the formation of stress granule (SG) in human periodontal ligament cells (hPDLCs) under stress, and its effect on the apoptosis, the expression of inflammatory factors, and the generation of reactive oxygen species (ROS) under lipopolysaccharide (LPS) stimulation. Methods: Sodium arsenite (SA) was used to induce SG formation in hPDLCs, and immunofluorescence staining was used to quantify SG formation. Live and dead cell staining, real-time quantitative polymerase chain reaction (RT-qPCR) and immunofluorescence staining were used to detect the localization and expression of cysteinyl aspartate-specific proteinase 3 (Caspase-3) in hPDLCs to evaluate the effect of SG formation on apoptosis. RT-qPCR was used to detect the mRNA expression changes of inflammatory-related cytokines interleukin 1β (IL-1β) and interleukin 6 (IL-6) during LPS stimulation. ROS detection was used to assess the effect of SG on LPS-induced ROS levels of hPDLCs. Results: SA successfully induced SG formation in hPDLCs. The results of immunofluorescence staining showed that Caspase-3 was localized in SG, which significantly inhibited the apoptosis of hPDLCs. RT-qPCR results indicated that SG could downregulate the mRNA relative expression of LPS-stimulated inflammatory cytokines in hPDLCs. ROS detection showed that SG inhibited LPS-induced ROS generation in hPDLCs. Conclusion: SG can protect hPDLCs from stress-induced apoptosis and inhibit the expression of LPS-triggered inflammatory-related factors and ROS generation.

Key words:

stress granule, apoptosis, periodontitis, reactive oxygen species

中图分类号:

R782

徐辉, 乔广艳, 苏俭生.

应激颗粒形成对人牙周膜细胞凋亡及对LPS刺激下炎症相关因子表达及ROS生成的影响[J]. 《口腔颌面外科杂志》, 2025, 35(3): 181-189.

XU Hui, QIAO Guangyan, SU Jiansheng.

Effects of stress granule formation on apoptosis of human periodontal ligament cells and the expression of inflammatory-related factors and ROS generation under LPS stimulation[J]. 《Journal of Oral and Maxillofacial Surgery》, 2025, 35(3): 181-189.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2025/V35/I3/181

参考文献

[1] Pan WY, Wang QX, Chen QM. The cytokine network involved in the host immune response to periodontitis[J]. Int J Oral Sci, 2019, 11(3): 30.

[2] Silva LM, Doyle AD, Greenwell-Wild T, et al. Fibrin is a critical regulator of neutrophil effector function at the oral mucosal barrier[J]. Science, 2021, 374(6575): eabl5450.

[3] Li ZX, Liu XH, Liu MJ. Stress granule homeostasis, aberrant phase transition, and amyotrophic lateral sclerosis[J]. ACS Chem Neurosci, 2022, 13(16): 2356-2370.

[4] Fujikawa D, Nakamura T, Yoshioka D, et al. Stress granule formation inhibits stress-induced apoptosis by selectively sequestering executioner caspases[J]. Curr Biol, 2023, 33(10): 1967-1981.e8.

[5] Coll RC, Schroder K, Pelegrín P. NLRP3 and pyroptosis blockers for treating inflammatory diseases[J]. Trends Pharmacol Sci, 2022, 43(8): 653-668.

[6] Samir P, Kesavardhana S, Patmore DM, et al. DDX3X acts as a live-or-die checkpoint in stressed cells by regulating NLRP3 inflammasome[J]. Nature, 2019, 573(7775): 590-594.

[7] Amen T, Kaganovich D. Stress granules inhibit fatty acid oxidation by modulating mitochondrial permeability[J]. Cell Rep, 2021, 35(11): 109237.

[8] Park JY, Shin OS. Stress granules inhibit coxsackievirus B3-mediated cell death via reduction of mitochondrial reactive oxygen species and viral extracellular release[J]. J Microbiol Biotechnol, 2023, 33(5): 582-590.

[9] Dai HQ, Wang GL, Cao WM, et al. Stress granules affect the sensitivity of renal cancer cells to sorafenib by sequestering and stabilizing COX-2 mRNA[J]. Oncol Lett, 2023, 25(6): 274.

[10] Paget M, Cadena C, Ahmad S, et al. Stress granules are shock absorbers that prevent excessive innate immune responses to dsRNA[J]. Mol Cell, 2023, 83(7): 1180-1196.e8.

[11] Protter DSW, Parker R. Principles and properties of stress granules[J]. Trends Cell Biol, 2016, 26(9): 668-679.

[12] Li TS, Zeng ZY, Fan CM, et al. Role of stress granules in tumorigenesis and cancer therapy[J]. Biochim Biophys Acta Rev Cancer, 2023, 1878(6): 189006.

[13] Yu QY, Ye LQ, Li HL. Molecular interaction of stress granules with Tau and autophagy in Alzheimer's disease[J].

Neurochem Int, 2022, 157: 105342.

[14] Chang AM, Kantrong N, Darveau RP. Maintaining homeostatic control of periodontal epithelial tissue[J]. Periodontol 2000, 2021, 86(1): 188-200.

[15] Baymiller M, Moon SL. Stress granules as causes and consequences of translation suppression[J]. Antioxid Redox Signal, 2023, 39(4-6): 390-409.

[16] Arimoto-Matsuzaki K, Saito H, Takekawa M. TIA1 oxidation inhibits stress granule assembly and sensitizes cells to stress-induced apoptosis[J]. Nat Commun, 2016, 7: 10252.

[17] Verma A, Sumi S, Seervi M. Heat shock proteins-driven stress granule dynamics: Yet another avenue for cell survival[J]. Apoptosis, 2021, 26(7-8): 371-384.

[18] Loos BG, Van Dyke TE. The role of inflammation and genetics in periodontal disease[J]. Periodontol 2000, 2020, 83(1): 26-39.

[19] Sczepanik FSC, Grossi ML, Casati M, et al. Periodontitis is an inflammatory disease of oxidative stress: We should treat it that way[J]. Periodontol 2000, 2020, 84(1): 45-68.

[20] Takahashi M, Higuchi M, Matsuki H, et al. Stress granules inhibit apoptosis by reducing reactive oxygen species production[J]. Mol Cell Biol, 2013, 33(4): 815-829.

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