芍药苷促β-防御素表达在念珠菌性口炎中的作用及机制研究

doi:10.13701/j.cnki.kqyxyj.2025.08.011

口腔医学研究 ›› 2025, Vol. 41 ›› Issue (8): 700-706.DOI: 10.13701/j.cnki.kqyxyj.2025.08.011

芍药苷促β-防御素表达在念珠菌性口炎中的作用及机制研究

肖丽君¹, 潘燕¹, 杨建堂^2*

1.滨州医学院附属烟台口腔医院黏膜科山东烟台 264000;
2.遵义医科大学附属口腔医院黏膜科贵州遵义 563006

收稿日期:2024-11-29 出版日期:2025-08-28 发布日期:2025-08-15
通讯作者: ^*杨建堂,E-mail:945810934@qq.com
作者简介:肖丽君(1996~ ),女,山东威海人,硕士,住院医师,研究方向:口腔黏膜病学。
基金资助:
国家自然科学基金(编号:81960202); 贵州省科技厅项目(黔科合基础-ZK[2023]一般534); 滨州医学院“口腔医学+X”院校融合创新项目(编号:KQRH2024MS006)

Role and Mechanism of Paeoniflorin Promoting β-Defensin Expression in Candidal Stomatitis

XIAO Lijun¹, PAN Yan¹, YANG Jiantang^2*

1. Department of Oral Mucosa, the Affiliated Yantai Stomatological Hospital, Binzhou Medical University, Yantai 264000, China;
2. Department of Oral Mucosa, the Affiliated Stomatological Hospital, Zunyi Medical University, Zunyi 563006, China

Received:2024-11-29 Online:2025-08-28 Published:2025-08-15

摘要/Abstract

摘要： 目的: 探讨芍药苷(paeoniflorin, PF)促进β-防御素表达在念珠菌性口炎中的作用及机制。方法: 采用不同浓度PF处理Leuk-1细胞24、48、72 h,CCK-8试剂盒(cell counting kit-8,CCK-8)法检测细胞活力,筛选后续实验PF浓度与作用时间;建立白色念珠菌(Candida albcians,C.a)感染口腔黏膜上皮细胞系Leuk-1的念珠菌性口炎体外模型,检测PF的抗菌能力;实时荧光定量PCR(quantitative reverse transcription polymerase chain reaction,RT-qPCR)、酶联免疫吸附实验(enzyme-linked immunosorbent assays,ELISA)检测PF处理口腔黏膜上皮细胞后β-防御素(human β-defensin, HBD-1、HBD-2、HBD-3)基因和蛋白的表达水平;Western blot检测核因子-κB(NF-κB)信号通路中磷酸化P65蛋白活化情况。结果: CCK-8结果显示,5 μmol/L、10 μmol/L为PF作用Leuk-1细胞的最佳浓度,可明显降低细胞模型中C.a的黏附率(P<0.05)。PF能够诱导Leuk-1中HBD-1、HBD-2、HBD-3基因和蛋白的表达,与空白组比较差异均有统计学意义(P<0.01),且C.a的生存率显著下降。PF作用细胞后,p-P65/P65蛋白相对表达量升高,阻断P65-NF-κB信号通路后,HBDs的基因、蛋白表达量明显降低(P<0.05),同时C.a的生存率显著下降(P<0.0001)。结论: PF可通过激活P65-NF-κB通路蛋白,有效诱导Leuk-1细胞合成分泌β-防御素,进而抑制C.a菌丝生长、控制C.a的感染与侵袭,为PF防治念珠菌性口炎和口腔黏膜疾病提供依据。

关键词: 芍药苷, 念珠菌性口炎, β-防御素

Abstract: Objective: To study the role and mechanism of paeoniflorin (PF) promoting β-defensin expression in candidal stomatitis. Methods: The model of candida albicans (C.a) infected Leuk-1 cells to detect the antibacterial ability of β-defensin was established. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbentAssays (ELISA) were used to detect the gene and protein expression level of β-defensin (HBD-1, HBD- 2, HBD-3). The expressions of phosphorylated P65 in NF-κB signaling pathway were detected by Western blot. Results: CCK-8 results showed that PF at the concentration of 5 μmol/L and 10 μmol/L was selected to treat Leuk-1 cells for 24 h for subsequent experiments which could significantly inhibit the adhesion rate of C.a to Leuk-1 (P<0.05). RT-qPCR and ELISA results showed that 5 μmol/L and 10 μmol/L concentrations of PF could induce Leuk-1 to express HBD-1, -2, and -3, which were statistically significant compared with the blank control group (P<0.01), and the survival rate of C.a under the action of PF decreased significantly. The expression of p-P65 protein was increased by PF. Under the action of inhibitors, the expression of HBD-1, -2, and -3 genes and proteins decreased significantly, the survival rate of C.a increased significantly (P<0.0001). Conclusion: PF induces Leuk-1 cells to secrete and express β-defensins and mucoprotein 1 mainly via P65-NF-κB signal pathway, which could inhibit the growth and control the infection and invasion of C.a, which provided a basis for PF to prevent and treat candida stomatitis and other oral mucosal disease.

Key words: paeoniflorin, candidal stomatitis, β-defensin

肖丽君, 潘燕, 杨建堂. 芍药苷促β-防御素表达在念珠菌性口炎中的作用及机制研究[J]. 口腔医学研究, 2025, 41(8): 700-706.

XIAO Lijun, PAN Yan, YANG Jiantang. Role and Mechanism of Paeoniflorin Promoting β-Defensin Expression in Candidal Stomatitis[J]. Journal of Oral Science Research, 2025, 41(8): 700-706.

参考文献

[1] Hellstein JW, Marek CL. Candidiasis: Red and white manifestations in the oral cavity [J]. Head Neck Pathol, 2019, 13(1): 25-32.
[2] Gan Y, Cui X, Ma T, et al. Paeoniflorin upregulates β-defensin-2 expression in human bronchial epithelial cell through the p38 MAPK, ERK, and NF-κB signaling pathways [J]. Inflammation, 2014, 37(5): 1468-1475.
[3] Millsop JW, Fazel N. Oral candidiasis [J]. Clin Dermatol, 2016, 34(4): 487-494.
[4] Höfs S, Mogavero S, Hube B. Interaction of Candida albicans with host cells: virulence factors, host defense, escape strategies, and the microbiota [J]. J Microbiol, 2016, 54(3): 149-169.
[5] Pereira R, Dos Santos Fontenelle RO, de Brito EHS, et al. Biofilm of Candida albicans: formation, regulation and resistance [J]. J Appl Microbiol, 2021, 131(1): 11-22.
[6] Swidergall M, Filler SG. Oropharyngeal Candidiasis: Fungal invasion and epithelial cell responses [J]. PLoS Pathog, 2017, 13(1): e1006056.
[7] Polke M, Hube B, Jacobsen ID. Candida survival strategies [J]. Adv Appl Microbiol, 2015, 91: 139-235.
[8] Moyes DL, Wilson D, Richardson JP, et al. Candidalysin is a fungal peptide toxin critical for mucosal infection [J]. Nature, 2016, 532(7597): 64-68.
[9] Tang SX, Moyes DL, Richardson JP, et al. Epithelial discrimination of commensal and pathogenic Candida albicans [J]. Oral Dis, 2016, 22 Suppl 1: 114-119.
[10] Prado-Montes de Oca E. Human beta-defensin 1: a restless warrior against allergies, infections and cancer [J]. Int J Biochem Cell Biol, 2010, 42(6): 800-804.
[11] Cruz Díaz LA, Gutiérrez Ortega A, Chávez álvarez RDC, et al. Regulatory SNP rs5743417 impairs constitutive expression of human β-defensin 1 and has high frequency in Africans and Afro-Americans [J]. Int J Immunogenet, 2020, 47(4): 332-341.
[12] Dornelas Figueira LM, Ricomini Filho AP, da Silva WJ, et al. Glucose effect on Candida albicans biofilm during tissue invasion [J]. Arch Oral Biol, 2020, 117: 104728.
[13] Fusco A, Savio V, Donniacuo M, et al. Antimicrobial peptides human beta-defensin-2 and -3 protect the gut during Candida albicans infections enhancing the intestinal barrier integrity: In vitro study [J]. Front Cell Infect Microbiol, 2021, 11: 666900.
[14] Miró MS, Caeiro JP, Rodriguez E, et al. Candida albicans modulates murine and human beta defensin-1 during vaginitis [J]. J Fungi (Basel), 2021, 8(1):20.
[15] Casaroto AR, da Silva RA, Salmeron S, et al. Candida albicans-cell interactions activate innate immune defense in human palate epithelial primary cells via nitric oxide (NO) and β-defensin 2 (hBD-2) [J]. Cells, 2019, 8(7):707.
[16] álvarez áH, Martínez Velázquez M, Prado Montes de Oca E. Human β-defensin 1 update: Potential clinical applications of the restless warrior [J]. Int J Biochem Cell Biol, 2018, 104: 133-137.
[17] Ryan LK, Diamond G. Modulation of human β-defensin-1 production by viruses [J]. Viruses, 2017, 9(6):153.
[18] Shelley JR, Davidson DJ, Dorin JR. The dichotomous responses driven by β-defensins [J]. Front Immunol, 2020, 11: 1176.
[19] Liang W, Diana J. The dual role of antimicrobial peptides in autoimmunity [J]. Front Immunol, 2020, 11: 2077.
[20] Tomalka J, Azodi E, Narra HP, et al. β-Defensin 1 plays a role in acute mucosal defense against Candida albicans [J]. J Immunol, 2015, 194(4): 1788-1795.
[21] Järvå M, Phan TK, Lay FT, et al. Human β-defensin 2 kills Candida albicans through phosphatidylinositol 4,5-bisphosphate-mediated membrane permeabilization [J]. Sci Adv, 2018, 4(7): eaat0979.
[22] Chang HT, Tsai PW, Huang HH, et al. LL37 and hBD-3 elevate the β-1,3-exoglucanase activity of Candida albicans Xog1p, resulting in reduced fungal adhesion to plastic [J]. Biochem J, 2012, 441(3): 963-970.
[23] Yoo YJ, Kwon I, Oh SR, et al. Antifungal effects of synthetic human beta-defensin-3-C15 peptide on Candida albicans-infected root dentin [J]. J Endod, 2017, 43(11): 1857-1861.
[24] Vylkova S, Li XS, Berner JC, et al. Distinct antifungal mechanisms: beta-defensins require Candida albicans Ssa1 protein, while Trk1p mediates activity of cysteine-free cationic peptides [J]. Antimicrob Agents Chemother, 2006, 50(1): 324-331.
[25] te Welscher YM, van Leeuwen MR, de Kruijff B, et al. Polyene antibiotic that inhibits membrane transport proteins [J]. Proc Natl Acad Sci U S A, 2012, 109(28): 11156-11159.
[26] Anderson TM, Clay MC, Cioffi AG, et al. Amphotericin forms an extramembranous and fungicidal sterol sponge [J]. Nat Chem Biol, 2014, 10(5): 400-406.
[27] Ostrosky-Zeichner L, Casadevall A, Galgiani JN, et al. An insight into the antifungal pipeline: selected new molecules and beyond [J]. Nat Rev Drug Discov, 2010, 9(9): 719-727.
[28] Odds FC, Brown AJ, Gow NA. Antifungal agents: mechanisms of action [J]. Trends Microbiol, 2003, 11(6): 272-279.
[29] Vila T, Sultan AS, Montelongo-Jauregui D, et al. Oral Candidiasis: A disease of opportunity [J]. J Fungi (Basel), 2020, 6(1):15.
[30] Arendrup MC, Perlin DS. Echinocandin resistance: an emerging clinical problem? [J]. Curr Opin Infect Dis, 2014, 27(6): 484-492.
[31] Zhang L, Wei W. Anti-inflammatory and immunoregulatory effects of paeoniflorin and total glucosides of paeony [J]. Pharmacol Ther, 2020, 207: 107452.
[32] Xiang Y, Zhang Q, Wei S, et al. Paeoniflorin: a monoterpene glycoside from plants of Paeoniaceae family with diverse anticancer activities [J]. J Pharm Pharmacol, 2020, 72(4): 483-495.
[33] Wang XZ, Xia L, Zhang XY, et al. The multifaceted mechanisms of Paeoniflorin in the treatment of tumors: State-of-the-Art [J]. Biomed Pharmacother, 2022, 149: 112800.
[34] Wu XX, Huang XL, Chen RR, et al. Paeoniflorin prevents intestinal barrier disruption and inhibits lipopolysaccharide (LPS)-induced inflammation in Caco-2 cell monolayers [J]. Inflammation, 2019, 42(6): 2215-2225.
[35] Fan Q, Guan X, Hou Y, et al. Paeoniflorin modulates gut microbial production of indole-3-lactate and epithelial autophagy to alleviate colitis in mice [J]. Phytomedicine, 2020, 79: 153345.
[36] Luo X, Wang X, Huang S, et al. Paeoniflorin ameliorates experimental colitis by inhibiting gram-positive bacteria-dependent MDP-NOD2 pathway [J]. Int Immunopharmacol, 2021, 90: 107224.

芍药苷促β-防御素表达在念珠菌性口炎中的作用及机制研究

Role and Mechanism of Paeoniflorin Promoting β-Defensin Expression in Candidal Stomatitis

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 3

编辑推荐

Metrics

[1]	肖丽君, 陈谦明, 吴玉琪, 杨建堂. 基于转录组测序分析芍药苷诱导下口腔黏膜上皮细胞差异表达基因[J]. 口腔医学研究, 2023, 39(6): 527-533.
[2]	谷悦, 张静, 梁蓓蓓, 杨佳迪, 王佳宁, 刘娜, 刘庆. 光活化消毒治疗小鼠急性假膜型念珠菌性口炎的效果观察[J]. 口腔医学研究, 2022, 38(8): 762-767.
[3]	李丽丽,陈斌,崔迪,姜苏,闫福华. 人β-防御素-3对小鼠牙周炎进展的保护作用[J]. 口腔医学研究, 2016, 32(9): 897-901.