Email Alert | RSS

北京大学学报（医学版） ›› 2021, Vol. 53 ›› Issue (4): 744-749. doi: 10.19723/j.issn.1671-167X.2021.04.021

• 论著 • 上一篇    下一篇

钙结合蛋白在健康牙周组织和实验性牙周炎组织的表达分布

郜洪宇1,2,孟焕新1,Δ(),侯建霞1,黄宝鑫1,李玮1   

  1. 1.北京大学口腔医学院·口腔医院,牙周科 国家口腔医学中心 国家口腔疾病临床医学研究中心 口腔数字化医疗技术和材料国家工程实验室,北京 100081
    2.天津医科大学口腔医院牙周科,天津 300070
  • 收稿日期:2019-12-11 出版日期:2021-08-18 发布日期:2021-08-25
  • 通讯作者: 孟焕新 E-mail:kqhxmeng@bjmu.edu.cn
  • 基金资助:
    国家自然科学基金(81570980);国家自然科学基金(81870773);天津市教委科研计划项目(2016YD16);天津医科大学口腔医院科研基金(2015YKYB02)

Expression and distribution of calprotectin in healthy and inflamed periodontal tissues

GAO Hong-yu1,2,MENG Huan-xin1,Δ(),HOU Jian-xia1,HUANG Bao-xin1,LI Wei1   

  1. 1. Department of Periodontology, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing 100081, China
    2. Department of Periodontology, Stomatological Hospital of Tianjin Medical University, Tianjin 300070, China
  • Received:2019-12-11 Online:2021-08-18 Published:2021-08-25
  • Contact: Huan-xin MENG E-mail:kqhxmeng@bjmu.edu.cn
  • Supported by:
    National Natural Foundation of China(81570980);National Natural Foundation of China(81870773);Science & Technology Development Fund of Tianjin Education Commission for Higher Education(2016YD16);Science Foundation of Tianjin Medical University Stomatological Hospital(2015YKYB02)

RICH HTML

 11   

PDF (PC)

140

摘要：

目的: 观察比格犬相对健康的牙周组织和实验性牙周炎的牙周组织标本中钙结合蛋白的表达分布和细胞定位,探讨其与牙周炎症的关系和可能发挥的作用。方法: 结扎法诱导建立比格犬下颌第二磨牙实验性牙周炎模型,对侧同名牙作为健康对照,诱导12周后处死,组织脱矿后常规制作连续切片,免疫组织化学法检测钙结合蛋白在比格犬健康和实验性牙周炎的组织标本中的表达和分布,明确细胞定位;免疫细胞化学法检测体外培养的原代人牙周组织细胞中钙结合蛋白的表达。结果: 在健康牙龈组织中,钙结合蛋白表达于牙龈上皮、中性粒细胞,且在结合上皮处呈强阳性表达;牙周炎症病损中,牙龈上皮细胞钙结合蛋白表达水平上调,存在强表达钙结合蛋白的中性粒细胞大量浸润,成纤维样细胞(牙龈结缔组织、牙周膜组织)、骨髓成纤维细胞、微血管内皮细胞存在钙结合蛋白的诱导表达;在体外培养的人牙周膜细胞、人牙龈成纤维细胞中均检测到钙结合蛋白的表达。结论: 钙结合蛋白组成性表达于牙龈上皮细胞、中性粒细胞,可能对维持牙周组织内环境稳定和完整性发挥重要作用。牙周炎症诱导牙龈成纤维细胞、牙周膜细胞、骨髓成纤维细胞和血管内皮细胞表达的钙结合蛋白可能在抗微生物感染、促炎症细胞迁移等方面发挥作用。

关键词: 钙结合蛋白, 牙周炎, 结合上皮, 牙龈成纤维细胞, 牙周膜细胞

Abstract:

Objective: Calprotectin, the heterdimer of S100A8 and S100A9, is the major cytoplasmic protein of neutrophils, which is also expressed or induced in gingival epithelial cells, activated mononuclear macrophages and vascular endothelial cells. Calprotectin is intimately associated with the initiation and progression of periodontitis, but the in vivo expression patterns of calprotectin in healthy and inflamed periodontal tissue are not fully understood. To observe the expression, distribution and cellular localization of calprotectin in the samples of healthy periodontal tissues and experimental periodontitis tissues of Beagles and to explore their relationship with periodontal inflammation and possible effect. Methods: Experimental periodontitis model was established by ligation around the mandibular second molar of the Beagle dogs, while the contralateral teeth were healthy controls. Induction duration was 12 weeks, before the dogs were executed. Tissue specimens were demineralized and serial sections were made conventionally. The in vivo expression of calprotectin in the healthy and inflamed periodontal tissues were examined by immunohistochemistry. The in vitro expression of calprotectin in human primary gingival fibroblasts (GFs) and periodontal ligament (PDL) cells were detected by immunocytochemistry. Results: Immunohistochemistry analysis indicated that calprotectin was expressed in gingival epithelial cells and infiltrated neutrophils in the healthy periodontium within the gingival epithelium, S100A8/A9 was most strongly expressed in the junctional epithelium, followed by surface epithelium, and least expressed in the sulcular epithelium. The S100A8/A9 expression levels were sharply defined at the junction between the junctional epithelium and the sulcular epithelium. In periodontal inflammatory lesions, the expression level of calprotectin in sulcular epithelium and junctional epithelium was up-regulated than that in the healthy gingival epithelium. Calprotectin was inducibly expressed in fibroblast-like cells in gingival connective tissue and periodontal ligament tissue, microvascular endothelial cells (ECs) and bone marrow fibroblasts under inflammatory conditions. Additionally, the expression of calprotectin in primary human GFs and PDL cells was confirmed by immunnocytochemistry staining. Conclusion: Constitutively expressed in neutrophils and gingival epithelial cells, and calprotectin might maintain the homeostasis and integrity of periodontium. Inflammation-induced expression of calprotectin in GFs, PDL cells, microvascular ECs and bone marrow fibroblasts might process anti-microbial function and promote leukocytes transmigration to defend the host against the microorganisms.

Key words: Calprotectin, Periodontitis, Junctional epithelium, Gingvial fibroblasts, Periodontal ligament cells.

中图分类号: 

  • R781.4

图1

S100A8/A9在健康及实验性牙周炎牙龈组织中的表达分布"

图2

S100A8/A9在健康及实验性牙周炎根分叉组织中的表达分布"

图3

S100A8/A9在人牙龈成纤维细胞、牙周膜细胞的表达"

[1] Hajishengallis G. Periodontitis: from microbial immune subversion to systemic inflammation [J]. Nat Rev Immunol, 2015, 15(1):30-44.
doi: 10.1038/nri3785 pmid: 25534621
[2] Page RC, Kornman KS. The pathogenesis of human periodontitis: an introduction [J]. Periodontol 2000, 1997, 14(1):9-11.
doi: 10.1111/prd.1997.14.issue-1
[3] Di Benedetto A, Gigante I, Colucci S, et al. Periodontal disease: linking the primary inflammation to bone loss [J/OL]. Clin Dev Immunol, 2013, 2013: 503754(2013-05-23)[2019-12-01]. https//www.hindawi.com/journals/ .
[4] Kolaczkowska E, Kubes P. Neutrophil recruitment and function in health and inflammation [J]. Nat Rev Immunol, 2013, 13(3):159-175.
doi: 10.1038/nri3399 pmid: 23435331
[5] Jin L. An update on innate defense molecules of human gingiva [J]. Periodontol 2000, 2011, 56(1):125-142.
doi: 10.1111/prd.2011.56.issue-1
[6] Goyette J, Geczy CL. Inflammation-associated S100 proteins: new mechanisms that regulate function [J]. Amino Acids, 2011, 41(4):821-842.
doi: 10.1007/s00726-010-0528-0
[7] Lundy FT, Chalk R, Lamey PJ, et al. Identification of MRP-8 (calgranulin A) as a major responsive protein in chronic periodontitis [J]. J Pathol, 2000, 192(4):540-544.
pmid: 11113873
[8] Lundy FT, Chalk R, Lamey PJ, et al. Quantitative analysis of MRP-8 in gingival crevicular fluid in periodontal health and disease using microbore HPLC [J]. J Clin Periodontol, 2001, 28(12):1172-1177.
pmid: 11737516
[9] Kido J, Nakamura T, Kido R, et al. Calprotectin in gingival crevicular fluid correlates with clinical and biochemical markers of periodontal disease [J]. J Clin Periodontol, 1999, 26(10):653-657.
pmid: 10522776
[10] Andersen E, Dessaix IM, Perneger T, et al. Myeloid-related protein (MRP8/14) expression in gingival crevice fluid in periodontal health and disease and after treatment [J]. J Periodontal Res, 2010, 45(4):458-463.
doi: 10.1111/j.1600-0765.2009.01257.x pmid: 20337885
[11] Ren XY, Xu L, Meng HX, et al. Family-based association analysis of S100A8 genetic polymorphisms with aggressive periodontitis [J]. J Periodontal Res, 2009, 44(2):184-192.
doi: 10.1111/j.1600-0765.2008.01103.x pmid: 19210342
[12] Li QY, Meng HX, Zhang L, et al. Correlation between single nucleotide polymorphisms in a calprotectin subunit gene and risk of periodontitis in a Chinese population [J]. Ann Hum Genet, 2007, 71(Pt 3):312-324.
doi: 10.1111/ahg.2007.71.issue-3
[13] Sun XJ, Meng HX, Shi D, et al. Analysis of plasma calprotectin and polymorphisms of S100A8 in patients with aggressive periodontitis [J]. J Periodontal Res, 2011, 46(3):354-360.
doi: 10.1111/j.1600-0765.2011.01350.x pmid: 21463326
[14] 孙晓军, 孟焕新, 释栋, 等. 侵袭性牙周炎患者血浆钙结合蛋白与血液中性粒细胞数相关性的研究 [J]. 中华口腔医学杂志, 2014, 49(11):649-651.
[15] Suryono, Kido J, Hayashi N, et al. Effect of porphyromonas gingivalis lipopolysaccharide, tumor necrosis factor-alpha, and interleukin-1beta on calprotectin release in human monocytes [J]. J Periodontol, 2003, 74(12):1719-1724.
doi: 10.1902/jop.2003.74.12.1719 pmid: 29539076
[16] Schlegel Gomez R, Langer P, Pelka M, et al. Variational expression of functionally different macrophage markers (27E10, 25F9, RM3/1) in normal gingiva and inflammatory periodontal disease [J]. J Clin Periodontol, 1995, 22(5):341-346.
pmid: 7541406
[17] Li W, Huang B, Liu K, et al. Up-regulated leptin in periodontitis promotes inflammatory cytokine expression in periodontal ligament cells [J]. J Periodontol, 2015, 86(7):917-926.
doi: 10.1902/jop.2015.150030
[18] Gao H, Hou J, Meng H, et al. Proinflammatory effects and mechanisms of calprotectin on human gingival fibroblasts [J]. J Periodontal Res, 2017, 52(6):975-983.
doi: 10.1111/jre.12465 pmid: 28643937
[19] Gao H, Zhang X, Zheng Y, et al. S100A9-induced release of interleukin (IL)-6 and IL-8 through toll-like receptor 4 (TLR4) in human periodontal ligament cells [J]. Mol Immunol, 2015, 67(2 Pt B):223-232.
doi: 10.1016/j.molimm.2015.05.014
[20] Hiroshima Y, Bando M, Kataoka M, et al. Regulation of antimicrobial peptide expression in human gingival keratinocytes by interleukin-1alpha [J]. Arch Oral Biol, 2011, 56(8):761-767.
doi: 10.1016/j.archoralbio.2011.01.004 pmid: 21316034
[21] Hsu K, Champaiboon C, Guenther BD, et al. Anti-infective protective properties of S100 calgranulins [J]. Antiinflamm Antiallergy Agents Med Chem, 2009, 8(4):290-305.
doi: 10.2174/187152309789838975
[22] Jonsson D, Nebel D, Bratthall G, et al. The human periodontal ligament cell: a fibroblast-like cell acting as an immune cell [J]. J Periodontal Res, 2011, 46(2):153-157.
doi: 10.1111/jre.2011.46.issue-2
[23] Ara T, Kurata K, Hirai K, et al. Human gingival fibroblasts are critical in sustaining inflammation in periodontal disease [J]. J Periodontal Res, 2009, 44(1):21-27.
doi: 10.1111/jre.2009.44.issue-1
[24] Nisapakultorn K, Ross KF, Herzberg MC. Calprotectin expression in vitro by oral epithelial cells confers resistance to infection by porphyromonas gingivalis [J]. Infect Immun, 2001, 69(7):4242-4247.
pmid: 11401960
[25] Yen T, Harrison CA, Devery JM, et al. Induction of the S100 chemotactic protein, CP-10, in murine microvascular endothelial cells by proinflammatory stimuli [J]. Blood, 1997, 90(12):4812-4821.
pmid: 9389698
[1] 朱小玲,李文静,王宪娥,宋文莉,徐莉,张立,冯向辉,路瑞芳,释栋,孟焕新. 细胞色素B-245α链及胆固醇酯转运蛋白基因多态性与广泛型侵袭性牙周炎易感性的关系[J]. 北京大学学报（医学版）, 2022, 54(1): 18-22.
[2] 徐欣然,霍芃呈,和璐,孟焕新,朱筠轩,靳东思奇. 伴与不伴糖尿病的牙周炎患者牙周基础治疗的疗效比较及其与白细胞水平的相关分析[J]. 北京大学学报（医学版）, 2022, 54(1): 48-53.
[3] 刘建,王宪娥,吕达,乔敏,张立,孟焕新,徐莉,毛铭馨. 广泛型侵袭性牙周炎患者牙根形态异常与相关致病基因的关联[J]. 北京大学学报（医学版）, 2021, 53(1): 16-23.
[4] 郜洪宇,徐菁玲,孟焕新,和璐,侯建霞. 牙周基础治疗对2型糖尿病伴慢性牙周炎患者红细胞、血小板相关指标的影响[J]. 北京大学学报（医学版）, 2020, 52(4): 750-754.
[5] 闫乐,王宪娥,詹雅琳,苗莉莉,韩烨,张楚人,岳兆国,胡文杰,侯建霞. 超声龈下清创联合手工根面平整术治疗重度牙周炎的临床效果[J]. 北京大学学报（医学版）, 2020, 52(1): 64-70.
[6] 张勇,刘畅,陈彬,陈帆,段晋瑜,张孟钧,焦剑. 糖尿病前期患者糖代谢异常与慢性牙周炎的相关性[J]. 北京大学学报（医学版）, 2020, 52(1): 71-76.
[7] 朱洁,李建红,袁婷婷,和璐,梁宇红. 绝经期妇女牙周状况与骨质密度的相关性分析[J]. 北京大学学报（医学版）, 2019, 51(6): 1115-1118.
[8] 石姝雯,孟洋,焦剑,李文静,孟焕新,栾庆先,王万春. 根分叉病变患牙经牙周非手术治疗后5年失牙状况及多因素分析[J]. 北京大学学报（医学版）, 2019, 51(5): 913-918.
[9] 杜仁杰,焦剑,周彦恒,施捷. 侵袭性牙周炎患者正畸前后的咬合变化[J]. 北京大学学报（医学版）, 2019, 51(5): 919-924.
[10] 刘园，栾庆先. 北京石景山社区中老年人群慢性牙周炎和颈动脉内膜中层厚度的相关性[J]. 北京大学学报(医学版), 2018, 50(2): 264-270.
[11] 张海东，张立，释栋，韩劼，闫夏，谢也斯，孟焕新. 锥形锁柱种植体用于因牙周炎缺牙患者修复的临床观察[J]. 北京大学学报(医学版), 2018, 50(2): 300-307.
[12] 张又文，辛天艺，焦剑，周彦恒，施捷. 慢性牙周炎的减数正畸治疗[J]. 北京大学学报(医学版), 2018, 50(2): 308-313.
[13] 李熠，徐莉，周彦恒，欧阳翔英，曹甜. 牙周正畸牙体联合治疗1例预后无望上前牙患者的长期疗效观察#br#[J]. 北京大学学报(医学版), 2017, 49(4): 740-744.
[14] 高丽，于晓潜，蔡宇. 丝线结扎及局部涂抹牙龈卟啉单胞菌对小鼠牙槽骨骨吸收的影响[J]. 北京大学学报(医学版), 2017, 49(1): 31-035.
[15] 沈潇，施捷，徐莉，焦剑，路瑞芳，孟焕新. 伴错牙合畸形的侵袭性牙周炎患者牙周-正畸联合治疗的临床评价[J]. 北京大学学报(医学版), 2017, 49(1): 60-066.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] 田增民, 陈涛, Nanbert ZHONG, 李志超, 尹丰, 刘爽. 神经干细胞移植治疗遗传性小脑萎缩的临床研究（英文稿）[J]. 北京大学学报（医学版）, 2009, 41(4): 456 -458 .
[2] 郭岩, 谢铮. 用一代人时间弥合差距——健康社会决定因素理论及其国际经验[J]. 北京大学学报（医学版）, 2009, 41(2): 125 -128 .
[3] 成刚, 钱振华, 胡军. 艾滋病项目自愿咨询检测的技术效率分析[J]. 北京大学学报（医学版）, 2009, 41(2): 135 -140 .
[4] 卢恬, 朱晓辉, 柳世庆, 郑杰, 邱晓彦. 白细胞介素2促进宫颈癌细胞系HeLaS3免疫球蛋白G的表达[J]. 北京大学学报（医学版）, 2009, 41(2): 158 -161 .
[5] 袁惠燕, 张苑, 范田园. 离子交换型栓塞微球及其载平阳霉素的制备与性质研究[J]. 北京大学学报（医学版）, 2009, 41(2): 217 -220 .
[6] 徐莉, 孟焕新, 张立, 陈智滨, 冯向辉, 释栋. 侵袭性牙周炎患者血清中抗牙龈卟啉单胞菌的IgG抗体水平的研究[J]. 北京大学学报（医学版）, 2009, 41(1): 52 -55 .
[7] 董稳, 刘瑞昌, 刘克英, 关明, 杨旭东. 氯诺昔康和舒芬太尼用于颌面外科术后自控静脉镇痛的比较[J]. 北京大学学报（医学版）, 2009, 41(1): 109 -111 .
[8] 祁琨, 邓芙蓉, 郭新彪. 纳米二氧化钛颗粒对人肺成纤维细胞缝隙连接通讯的影响[J]. 北京大学学报（医学版）, 2009, 41(3): 297 -301 .
[9] Jian-wei GU, Emily YOUNG, Zhi-jun PAN, Kevan B. TUCKER, Megan SHPARAGO, Min HUANG, Amelia Purser BAILEY. SD大鼠长期高盐饮食可导致其高血压并改变肾细胞因子基因表达谱[J]. 北京大学学报（医学版）, 2009, 41(5): 505 -515 .
[10] 李宏亮*, 安卫红*, 赵扬玉, 朱曦. 妊娠合并高脂血症性胰腺炎行血液净化治疗1例[J]. 北京大学学报（医学版）, 2009, 41(5): 599 -601 .
ISSN 1671-167X
CN 11-4691/R
主管单位：中华人民共和国教育部
主办单位：北京大学
地址： 北京市海淀区学院路38号 北京大学医学部 《北京大学学报（医学版）》
邮编：100191
电话：010-82801551
Email： xbbjb2@bjmu.edu.cn

《北京大学学报（医学版）》
微信公众号
版权所有 © 2018 《北京大学学报（医学版）》编辑部