国际口腔医学杂志

国际口腔医学杂志 ›› 2026, Vol. 53 ›› Issue (4): 513-525.doi: 10.7518/gjkq.2026128

• 论著 • 上一篇    

植物乳杆菌‑SLH对实验性牙周炎的治疗效果评价

舒路红1,2(),李航基雲1,2,张婷3,4,赵艳3,4,崔古贞3,4,洪伟1,3,廖健1,2()   

  1. 1.贵州医科大学口腔医学院/附属口腔医院 贵阳 550004
    2.贵州医科大学口腔疾病研究重点实验室 贵阳 550004
    3.贵州医科大学地方病与少数民族疾病教育部重点实验室 贵阳 550004
    4.贵州医科大学基础医学院微生物学教研室 贵阳 550025
  • 收稿日期:2025-06-09 修回日期:2026-02-11 出版日期:2026-07-01 发布日期:2026-06-25
  • 通讯作者: 廖健
  • 作者简介:舒路红，硕士，Email：1935679099@qq.com
  • 基金资助:
    国家自然科学基金(82260198);国家自然科学基金(32170134);贵州医科大学优秀青年人才培养计划(贵医大优秀青年人才（2022）101);贵州省科技计划项目（黔科合基础-ZK［2022］一般401）

Evaluation of the therapeutic effect of Lactobacillus plantarum SLH on experimental periodontitis

Luhong Shu1,2(),Hangjiyun Li1,2,Ting Zhang3,4,Yan Zhao3,4,Guzhen Cui3,4,Wei Hong1,3,Jian Liao1,2()   

  1. 1.School of Stomatology/Stomatological Hospital of Guizhou Medical University, Guiyang 550004, China
    2.Key Laboratory of Oral Disease Research, Guizhou Medical University, Guiyang 550004, China
    3.Key Laboratory of Endemic and Ethnic Diseases of the Ministry of Education, Guizhou Medical University, Guiyang 550004, China
    4.Dept. of Microbiology, School of Basic Medical Sciences, Guizhou Medical University, Guiyang 550025, China
  • Received:2025-06-09 Revised:2026-02-11 Online:2026-07-01 Published:2026-06-25
  • Contact: Jian Liao
  • Supported by:
    National Natural Science Foundation of China(82260198);Outstanding Young Ta-lent Training Program of Guizhou Medical University [Outstanding Young Talents at Guizhou Medical University (2022)101];Science and Technology Plan Project in Guizhou Province (Qiankehe Foundation-ZK[2022] General 401);National Natural Science Foundation of China (82260198, 32170134); Outstanding Young Ta-lent Training Program of Guizhou Medical University [Outstanding Young Talents at Guizhou Medical University (2022)101]; Science and Technology Plan Project in Guizhou Province (Qiankehe Foundation-ZK[2022] General 401)

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摘要：

目的 研究临床分离植物乳杆菌-SLH对实验性牙周炎的治疗和缓解作用。 方法 从健康婴儿粪便样品中分离纯化出1株植物乳杆菌-SLH为实验对象，革兰染色后进行形态学观察，绘制其生长曲线和产酸曲线，通过牛津杯琼脂扩散法检测菌体及发酵上清液对具核梭杆菌和牙龈卟啉单胞菌的抑菌活性，结晶紫染色法检测该菌株对牙周生物膜形成的抑制能力，并对菌株的溶菌酶耐受能力、体外抗炎能力进行评估。再用4-0丝线结扎法构建实验性牙周炎大鼠模型，将24只大鼠随机分配为正常对照组、牙周炎组和植物乳杆菌-SLH组，每组8只。用磷酸盐缓冲盐水（PBS）配置 2% 羧甲基纤维素（CMC）溶液作为植物乳杆菌-SLH的重悬溶剂施用于大鼠造模位点的牙周袋内，在正常对照组和牙周炎组分别注射同等量2% CMC的PBS，连续7周给药。给药结束后，对各组大鼠龈沟出血指数及牙周探诊深度进行检测和牙周组织病理学评估。 结果 成功分离纯化得到1株益生菌，经16S rDNA测序鉴定后命名为植物乳杆菌-SLH。抑菌实验及生物膜形成实验表明，该菌株的发酵上清液对牙周致病菌牙龈卟啉单胞菌和具核梭杆菌有抑菌活性（P<0.05）。溶菌酶耐受实验表明，植物乳杆菌-SLH具有耐受较高（3 mg/mL）溶菌酶的能力。另外，植物乳杆菌-SLH可降低炎症细胞模型中白细胞介素（IL）-1β、肿瘤坏死因子-α和IL-6炎症因子的表达（P<0.000 1）。与对照组比较，牙周炎组大鼠龈沟出血指数、牙周探诊深度增加（P<0.000 1）。植物乳杆菌-SLH组与牙周炎组比较，其龈沟出血指数、牙周探诊深度降低（P<0.000 1）。与对照组相比，牙周炎组体重减少（P<0.05），植物乳杆菌-SLH组与牙周炎组比较体重增加（P<0.000 1）。Micro-CT结果显示，牙周炎组骨吸收明显，植物乳杆菌-SLH组比牙周炎组骨吸收减少（P<0.001）。苏木精-伊红染色结果显示，植物乳杆菌-SLH组大鼠牙周组织炎症明显减轻。 结论 植物乳杆菌-SLH对实验性牙周炎具有显著的缓解作用，具有辅助治疗牙周炎的潜力。

关键词: 牙周炎, 植物乳杆菌, 牙龈卟啉单胞菌, 具核梭杆菌, 动物模型, 炎症反应

Abstract:

Objective　This study aimed to investigate the therapeutic potential of clinically isolated Lactobacillus plantarum SLH in experimental periodontitis. Methods　A strain of Lactobacillus plantarum SLH, isolated and purified from fecal samples of healthy infants, served as the experimental subject. Following Gram staining for morphological observation, its growth curve and acid production curve were plotted. The antibacterial activity of the bacterial cells and the cell-free fermentation supernatant against Fusobacterium nucleatum and Porphyromonas gingivalis was assessed using Oxford cup agar diffusion assay. The inhibitory effect of this strain on periodontal biofilm formation was evaluated via crystal violet staining assay. Furthermore, the strain’s tolerance to lysozyme and its in vitro anti-inflammatory capacity were determined. Subsequently, an experimental periodontitis model was established in rats by using ligature-induced method with 4-0 silk sutures. Twenty-four rats were randomly allocated into three groups (n=8 per group): a healthy control group, a periodontitis group, and an Lactobacillus plantarum SLH treatment group. A suspension of Lactobacillus plantarum SLH was prepared in phosphate buffered saline (PBS) containing 2% sodium carboxymethyl cellulose (CMC) as a vehicle. This suspension was administered locally into the periodontal pockets at the ligation sites in the treatment group. The healthy control and periodontitis groups received local injections of an equal volume of the vehicle (2% CMC in PBS) only. This treatment regimen continued for 7 consecutive weeks. Upon completion of the administration period, clinical periodontal parameters, including the gingival sulcus bleeding index and probing pocket depth, were measured for all rats, followed by histopathological evaluation of the periodontal tissues. Results　A probiotic strain was successfully isolated and purified. Following identification by 16S rDNA sequencing, it was designated as Lactobacillus plantarum SLH. Antibacterial and biofilm formation assays demonstrated that the cell-free fermentation supernatant of this strain exhibited significant inhibitory activity against the periodontal pathogens Porphyromonas gingivalis and Fusobacterium nucleatum (P<0.05). Lysozyme tolerance test indicated that Lactobacillus plantarum SLH could withstand a relatively high concentration (3 mg/mL) of lysozyme. Furthermore, Lactobacillus plantarum SLH significantly reduced the expression levels of the inflammatory cytokines interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and IL-6 in an inflammatory cell model (P<0.000 1). In the animal experiment, compared with the control group, the periodontitis group showed significant increases in the gingival sulcus bleeding index and probing pocket depth (P<0.000 1). Administration of Lactobacillus plantarum SLH resulted in a significant reduction in sulcus bleeding index (SBI) and probing depth (PD) compared with the treatment of periodontitis groups (P<0.000 1). The body weight of the periodontitis group decreased compared with that of the controls (P<0.05), and the Lactobacillus plantarum SLH treatment group showed a significant increase in body weight compared with the periodontitis group (P<0.000 1). Micro-computed tomography analysis revealed pronounced alveolar bone resorption in the periodontitis group. This bone loss was significantly attenuated in the Lactobacillus plantarum SLH treatment group compared with the periodontitis group (P<0.001). Histopathological examination using hematoxylin and eosin staining showed a marked alleviation of inflammatory cell infiltration and tissue damage in the periodontal tissues of rats treated with Lactobacillus plantarum SLH. Conclusion　Lactobacillus plantarum SLH exerts significant ameliorative effects on experimental periodontitis and holds potential as an adjunctive therapy for periodontal disease.

Key words: periodontitis, Lactobacillus plantarum, Porphyromonas gingivalis, Fusobacterium nucleatum, animal model, inflammatory response

中图分类号: 

  • R781.4+2

表 1

引物序列"

基因引物序列（5’—3’）
β-actin

F：AGGTCGGTGTGAACGGATTTG

R：TGTAGACCATGTAGTTGAGGTCA

IL-6

F：AGTCCGGAGAGGAGACTTC

R：AGTCCGGAGAGGAGACTTC

IL-1β

F：ACAAAATACCTGTGGCCTTGGG

R：TTGTTTCCCAGGAAGACAGGCT

TNF-α

F：AGAAAGCATGATCCGCGACGT

R：AGAAAGCATGATCCGCGACGT

图 1

植物乳杆菌-SLH的菌落形态及革兰染色A：菌落形态；B：革兰染色 × 1 000。"

图 2

植物乳杆菌-SLH 16S rDNA系统发育树绿色圆点所标注为本实验分离的植物乳杆菌-SLH菌株。"

图 3

植物乳杆菌-SLH的生长曲线及产酸曲线A：生长曲线；B：产酸曲线。"

图 4

植物乳杆菌-SLH菌体及发酵上清液的抑菌效果A：对具核梭杆菌的抑菌活性；B：对牙龈卟啉单胞菌的抑菌活性；①氯己定，②植物乳杆菌-SLH上清液，③MRS培养基，④植物乳杆菌-SLH菌体。"

表 2

植物乳杆菌-SLH菌体及上清液对具核梭杆菌及牙龈卟啉单胞菌的抑菌活性"

组别抑制具核梭杆菌的抑菌圈直径/mm抑制牙龈卟啉单胞菌的抑菌圈直径/mm
氯己定32.43±2.5335.75±1.23
植物乳杆菌-SLH菌体--
MRS培养基15.03±1.4915.27±0.29
植物乳杆菌-SLH上清液26.20±1.98***31.77±2.63****

图 5

抑制牙菌斑生物膜形成的测定A：结晶紫染色观察各处理组生物膜形态；B：各组生物膜形成的OD值测定。****：P<0.000 1。"

图 6

植物乳杆菌-SLH在不同溶菌酶浓度下的生长密度"

图 7

植物乳杆菌-SLH对细胞炎症因子mRNA表达的影响A：IL-1β基因相对表达量；B：IL-6基因相对表达量；C：TNF-α基因相对表达量。****：P<0.000 1。"

图 8

大鼠牙周炎模型的建立和牙周临床指标检测A：大鼠4-0丝线结扎造模；B：正常对照组牙龈组织大体观察情况；C：牙周炎组牙龈组织大体观察情况；D：植物乳杆菌-SLH组牙龈组织大体观察情况；E：大鼠体重变化；F：SBI、PD变化。**P<0.01；****：P<0.000 1。"

图 9

牙周组织HE染色结果和Micro-CT扫描结果分析A：大鼠牙周组织HE染色结果（AB为牙槽骨；FM为第一磨牙；SM为第二磨牙）；B：大鼠颌骨三维重建图及冠状面展示，依次为3组大鼠上颌第二磨牙牙槽骨丧失高度标记及冠状面骨量丧失情况，红色线段所示为CEJ-ABC的距离，标尺为2 mm；C：牙槽骨丧失的定量分析。*：P<0.05；***：P<0.001；****：P<0.000 1。"

[1] Zhang SP, Yu N, Arce RM. Periodontal inflammation: integrating genes and dysbiosis[J]. Periodontol 2000, 2020, 82(1): 129-142.
[2] Jiao J, Jing WD, Si Y, et al. The prevalence and severity of periodontal disease in mainland China: data from the Fourth National Oral Health Survey (2015—2016)[J]. J Clin Periodontol, 2021, 48(2): 168-179.
[3] Ebersole JL, Dawson Ⅲ D, Emecen-Huja P, et al. The periodontal war: microbes and immunity[J]. Pe-riodontol 2000, 2017, 75(1): 52-115.
[4] Lamont RJ, Koo H, Hajishengallis G. The oral microbiota: dynamic communities and host interactions[J]. Nat Rev Microbiol, 2018, 16(12): 745-759.
[5] Abdulkareem AA, Al-Taweel FB, Al-Sharqi AJB, et al. Current concepts in the pathogenesis of periodontitis: from symbiosis to dysbiosis[J]. J Oral Micro-biol, 2023, 15(1): 2197779.
[6] Tonetti MS, Van Dyke TE, GroupWorking 1 of the Joint EFP/AAP Workshop. Periodontitis and atherosclerotic cardiovascular disease: consensus report of the Joint EFP/AAP Workshop on Periodontitis and Systemic Diseases[J]. J Clin Periodontol, 2013, 40(s14): S24-S29.
[7] Baeza M, Morales A, Cisterna C, et al. Effect of periodontal treatment in patients with periodontitis and diabetes: systematic review and meta-analysis[J]. J Appl Oral Sci, 2020, 28: e20190248.
[8] Hajishengallis G, Chavakis T. Local and systemic mechanisms linking periodontal disease and inflammatory comorbidities[J]. Nat Rev Immunol, 2021, 21(7): 426-440.
[9] Bui FQ, Almeida-da-Silva CLC, Huynh B, et al. Association between periodontal pathogens and syste-mic disease[J]. Biomed J, 2019, 42(1): 27-35.
[10] Slots J. Concise evaluation and therapeutic guidelines for severe periodontitis: a public health perspective[J]. Periodontol 2000, 2022, 90(1): 262-265.
[11] Wei Y, Deng YX, Ma ST, et al. Local drug delivery systems as therapeutic strategies against periodontitis: a systematic review[J]. J Control Release, 2021, 333: 269-282.
[12] Gibson GR, Hutkins R, Sanders ME, et al. Expert consensus document: the International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics[J]. Nat Rev Gastroenterol Hepatol, 2017, 14(8): 491-502.
[13] Król-Górniak A, Pomastowski P, Railean-Plugaru V, et al. The study of the molecular mechanism of Lactobacillus paracasei clumping via divalent metal ions by electrophoretic separation[J]. J Chromatogr A, 2021, 1652: 462127.
[14] Khalaf H, Nakka SS, Sandén C, et al. Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis [J]. BMC Microbiol, 2016, 16(1): 188.
[15] Sulijaya B, Yamada-Hara M, Yokoji-Takeuchi M, et al. Antimicrobial function of the polyunsaturated fatty acid KetoC in an experimental model of periodontitis[J]. J Periodontol, 2019, 90(12): 1470-1480.
[16] 张晓旭, 刘欢, 肖苗, 等. 益生菌代谢产物对病原菌的抑菌作用研究进展[J]. 食品与发酵工业, 2023, 49(8): 297-302.
Zhang XX, Liu H, Xiao M, et al. Research progress in the antibacterial effect of probiotic metabolites on pathogens[J]. Food Ferment Ind, 2023, 49(8): 297-302.
[17] Plaza-Diaz J, Ruiz-Ojeda FJ, Gil-Campos M, et al. Mechanisms of action of probiotics[J]. Adv Nutr, 2019, 10: S49-S66.
[18] Abe-Yutori M, Chikazawa T, Shibasaki K, et al. Decreased expression of E-cadherin by Porphyromonas gingivalis-lipopolysaccharide attenuates epithelial barrier function[J]. J Periodontal Res, 2017, 52(1): 42-50.
[19] Yamada M, Takahashi N, Matsuda Y, et al. A bacte-rial metabolite ameliorates periodontal pathogen-induced gingival epithelial barrier disruption via GPR-40 signaling[J]. Sci Rep, 2018, 8(1): 9008.
[20] 翟佳琳, 赵景娜, 王丹丹, 等. 具有抑菌活性植物乳杆菌的筛选及抑菌物质特性的研究[J]. 食品与发酵工业, 2022, 48(13): 84-90.
Zhai JL, Zhao JN, Wang DD, et al. Screening of Lactiplantibacillus plantarum with bacteriostatic activity and study on properties of bacteriostatic substances[J]. Food Ferment Ind, 2022, 48(13): 84-90.
[21] 夏博园, 李艳, 丁旭, 等. 过氧化物酶体增殖物激活受体γ共激活因子-1α在牙周炎诱发大鼠肝损伤中的作用研究[J]. 华西口腔医学杂志, 2021, 39(5): 518-523.
Xia BY, Li Y, Ding X, et al. Effect of peroxisome proliferator-activated receptor-γ coactivator-1α on liver injury induced by periodontitis in rats[J]. West China J Stomatol, 2021, 39(5): 518-523.
[22] 余意, 王超越, 吴正钧, 等. 四株乳杆菌作为口腔益生菌的特性研究[J]. 食品与发酵工业, 2021, 47(15): 77-83.
Yu Y, Wang CY, Wu ZJ, et al. Potentials of Lactobacillus strains as oral probiotics candidates[J]. Food Ferment Ind, 2021, 47(15): 77-83.
[23] Shao Y, Garcia-Mauriño C, Clare S, et al. Primary succession of Bifidobacteria drives pathogen resistance in neonatal microbiota assembly[J]. Nat Microbiol, 2024, 9(10): 2570-2582.
[24] Chen ZH, Leng XY, Zhou F, et al. Screening and identification of probiotic Lactobacilli from the infant gut microbiota to alleviate lead toxicity[J]. Probiotics Antimicrob Proteins, 2023, 15(4): 821-831.
[25] Yu AO, Goldman EA, Brooks JT, et al. Strain diversity of plant-associated Lactiplantibacillus plantarum [J]. Microb Biotechnol, 2021, 14(5): 1990-2008.
[26] Van Holm W, Lauwens K, De Wever P, et al. Probio-tics for oral health: do they deliver what they pro-mise[J]. Front Microbiol, 2023, 14: 1219692.
[27] Guimarães A, Venancio A, Abrunhosa L. Antifungal effect of organic acids from lactic acid bacteria on Penicillium nordicum [J]. Food Addit Contam Part A, 2018, 35(9): 1803-1818.
[28] 杨颖, 陈卫, 张灏, 等. 植物乳杆菌HO-69的口腔益生性质研究[J]. 华西口腔医学杂志, 2008, 26(5): 482-485, 489.
Yang Y, Chen W, Zhang H, et al. Probiotic characte-ristics of Lactobacillus plantarum HO-69 applied in oral cavity[J]. West China J Stomatol, 2008, 26(5): 482-485, 489.
[29] Yoon JH, Oh MS, Lee SY. Effectiveness of organic acids for inactivating pathogenic bacteria inoculated in laboratory media and foods: an updated minireview[J]. Food Sci Biotechnol, 2024, 33(12): 2715-2728.
[30] 张军, 田子罡, 王建华, 等. 有机酸抑菌分子机理研究进展[J]. 畜牧兽医学报, 2011, 42(3): 323-328.
Zhang J, Tian ZG, Wang JH, et al. Advances in antimicrobial molecular mechanism of organic acids[J]. Acta Vet Zootechnica Sin, 2011, 42(3): 323-328.
[31] Wang SY, Nie ZY, Zhu L, et al. Probiotic characte-ristics and the anti-inflammatory effects of Lactiplantibacillus plantarum Z22 isolated from naturally fermented vegetables[J]. Microorganisms, 2024, 12(11): 2159.
[32] de Almeida Brandão D, Spolidorio LC, Johnson F, et al. Dose-response assessment of chemically modified curcumin in experimental periodontitis[J]. J Pe-riodontol, 2019, 90(5): 535-545.
[33] Pudgar P, Povšič K, Čuk K, et al. Probiotic strains of Lactobacillus brevis and Lactobacillus plantarum as adjunct to non-surgical periodontal therapy: 3-month results of a randomized controlled clinical trial[J]. Clin Oral Investig, 2021, 25(3): 1411-1422.
[34] Lam RS, O’Brien-Simpson NM, Lenzo JC, et al. Macrophage depletion abates Porphyromonas gingivalis-induced alveolar bone resorption in mice[J]. J Immunol, 2014, 193(5): 2349-2362.
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