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牙龈炎和牙周炎患者龈沟液内一氧化氮水平变化

2017-07-06周燕平覃沅华曲晓东

现代仪器与医疗 2017年3期
关键词:牙龈炎一氧化氮牙周炎

周燕平+覃沅华+曲晓东

[摘 要] 目的:观察牙龈炎和牙周炎患者龈沟液内一氧化氮(NO)水平变化。方法:随机抽取牙龈炎组、牙周炎组及正常组。使用硝酸还原酶法测定3组受试者龈沟液内NO水平,并检测牙龈炎组、牙周炎组患者治疗前后牙周临床指标、龈沟液内NO水平变化。结果:正常组龈沟液NO水平低于牙龈炎组,牙龈炎组龈沟液NO水平低于牙周炎组,差异有统计学意义(P<0.05)。两组患者治疗后龈沟液内NO水平有所下降仍高于正常组,差异有统计学意义(P<0.05)。牙龈炎组、牙周炎组患者牙周临床指标均较治疗前降低,牙龈炎组治疗后牙周探诊深度、附着丧失低于牙周炎组,差异有统计学意义(P<0.05)。结论:健康人群龈沟液内亦可检出NO,但水平较低,NO水平的上升伴随着牙周疾病的发生、发展,且与牙周疾病炎症程度具有一定关联。

[关键词] 牙龈炎;牙周炎;龈沟液;一氧化氮

中图分类号:R781.05 文献标识码:A 文章编号:2095-5200(2017)03-090-03

DOI:10.11876/mimt201703037

[Abstract] Objective: This study was conducted to observe the changes of nitric oxide (NO) level in gingival crevicular fluid of patients with gingivitis and periodontitis. Methods: Gingivitis group, periodontitis group and normal group were randomly selected. The levels of NO in gingival crevicular fluid of all patients in the three groups were measured by nitric acid reductase method. The changes of NO level in gingival crevicular fluid were observed before and after treatment in gingivitis group and periodontitis group. Results: The NO levels of the normal group in the gingival crevicular fluid was lower than that in the gingivitis group, and the NO levels of gingivitis group in gingival crevicular fluid was lower than that in the periodontitis group, the difference was statistically significant (P<0.05). The NO levels in gingival crevicular fluid of gingivitis group and periodontitis group were decreased after treatment, but were still higher than that in the normal group, the difference was statistically significant (P<0.05). The periodontal clinical indexes of gingivitis group and periodontitis group were lower than those before treatment, and periodontal probing depth and detachment loss in periodontitis group were both significantly lower than those in periodontitis group (P<0.05). Conclusions: NO can be detected in the gingival crevicular fluid of healthy people, but the levels of NO is related to the occurrence and development of periodontal disease, and it is related to the degree of inflammation of periodontal disease.

[Key words] gingivitis; periodontitis; gingival crevicular fluid; nitric oxide

牙周病是一種菌斑感染导致的牙齿支持组织慢性炎症性病变,包括牙龈炎、牙周炎等,以牙周支持组织破坏为主要表现[1]。作为一种化学性质极其活泼的自由基,一氧化氮(NO)不仅参与了循环、消化、神经、免疫等多个生理过程,也在牙周组织病理改变过程中扮演了重要角色[2]。龈沟液为牙龈结缔组织渗入到龈沟内的体液,检测牙龈炎和牙周炎患者龈沟液内NO水平变化,了解其引发牙周病的机制,对于疾病的诊断、评估与治疗均具有价值[3]。为此,本研究进行了对照分析,现报道如下。

1 资料与方法

1.1 一般资料

随机抽取2012年3月—2015年3月41例牙龈炎和57例慢性牙周炎患者,入组前6个月内无牙周病治疗史、抗生素使用史;选取同期35名健康体检者,均经全面检查排除口腔粘膜与全身代谢性疾病[4],纳入正常组。本临床研究已征得我院医学伦理委员会批准,受试者均知情同意并签署知情同意书。

1.2 检测方法

1.2.1 龈沟液内NO检测 取受试者双侧上下前后牙各1颗,其中前牙取中切牙(若缺失则取侧切牙),后牙取第一恒磨牙(若缺失则取第二恒磨牙),清除牙垢、牙石,漱口,干燥牙面,覆盖棉卷隔湿,采用袋底法提取龈沟液[5],取样部位包括颊、腭舌侧近中、远中牙周袋内,各取样部位留置30 s,确保吸潮纸未受血液或唾液污染,立即将其置于EP管内,3 min后重复上次操作,相同部位吸潮纸置于同一EP管内,-70℃液氮保存,待测。检测时将标本取出,4℃解冻,以300 μL蒸馏水浸没,3000 r/min离心5 min,留取上清液,以硝酸酶还原法实施NO检测[6]。

1.2.2 相关牙周检查及指标记录 记录3组受试者牙龈指数、出血指数、牙周探诊深度,按照文献[7] 进行评分。若探诊深度超过3 mm且袋底位于牙周袋根部则判断发生附着丧失,若牙龈无退缩,附着丧失=牙周探诊深度-釉牙骨质界到龈缘距离,若牙龈存在退缩,则附着丧失=牙周探诊深度+釉牙骨质界到龈缘距离。检测区域包括舌侧边缘龈、颊侧近中龈乳头、颊侧边缘龈及颊侧远中龈乳头,各区域检查结果平均值为最终结果。

1.3 治疗方案

牙龈炎、牙周炎患者均接受相关治疗,包括菌斑控制、龈上洁治、龈下刮治、咬合调整、牙周袋处理等[8],并于治疗后再次检测龈沟液内NO、相关牙周指标,观察患者上述指标变化情况。

2 结果

2.1 NO检测结果

正常组、牙龈炎组、牙周炎组受试者龈沟液内NO水平分别为(0.63±0.14)μmol/L、(5.19±1.33)μmol/L、

(7.26±2.08)μmol/L,正常组龈沟液NO水平低于牙龈炎组,牙龈炎组龈沟液NO水平低于牙周炎组,差异有统计学意义(P<0.05)。牙龈炎组治疗后龈沟液内NO水平为(2.87±0.45)μmol/L,低于牙周炎组患者治疗后的(4.68±1.24)μmol/L,但两组患者治疗后龈沟液内NO水平仍高于正常组,差异有统计学意义(P<0.05)。

2.2 牙周临床指标

正常组牙龈指数、出血指数、牙周探诊深度、附着丧失均低于牙龈炎组,牙龈炎组上述牙周临床指标均低于牙周炎组,差异有统计学意义(P<0.05)。

牙龈炎组、牙周炎组患者牙周临床指标均较治疗前降低,牙龈炎组治疗后牙周探诊深度、附着丧失低于牙周炎组,差异有统计学意义(P<0.05)。见表1。

3 讨论

NO是左型精氨酸在一氧化氮合酶作用下生成的小分子气体自由基,具有极其活泼的化学性质,可参与机体循环、消化、免疫等多种系统病理生理过程[9]。自国外学者首次于唾液中检出NO存在以来,NO在牙周组织疾病中作用时有报道 [10]。

本研究健康人群龈沟液亦可检出NO,但水平较低,说明NO主要来源于龈沟液,并可参与牙周微循环调节、维持牙周正常菌群平衡、抗微生物感染等作用环节[11-12]。而牙龈炎和牙周炎患者龈沟液内NO水平明显上升,考虑与患者牙周组织均存在明显炎症有关,此时大量聚集的炎症细胞可增强诱导型NO(iNO)表达,进而引发NO水平上升[13]。既往研究也表明,随着炎症程度的加剧,牙周组织疾病患者龈沟液内NO水平呈上升趋势[14]。

在牙周指标的观察中,可以发现,NO与牙周指标的变化趋势相同,即随着龈沟液内NO水平的上升,患者牙龈指数、出血指数、牙周探诊深度、附着丧失逐渐增加,说明过量的NO主要扮演细胞毒效应分子和炎性产物,从而造成内环境稳态调节障碍、血管内皮破坏、微循环异常,引发疾病的进一步发展[15],其机制是炎症诱导的NO大量生成可导致自然杀伤细胞活化,发挥病原微生物和肿瘤细胞杀伤作用,但过量NO可造成扩血管因子与缩血管因子间平衡被打破,进而造成多重级联信号活化、引发炎症瀑布级联反应,扩大信号范围,加剧炎症进展[16];龈沟液内NO主要来自于巨噬细胞、淋巴细胞、中性粒白细胞等,而大量生成的NO可诱导上述细胞自身凋亡加剧,从而造成机体调控炎癥反应能力下降、抗微生物防御能力不足,故牙龈炎和牙周炎均不存在自愈性,且随着时间的推移,患者症状逐渐加重[17]。

经规范化治疗,牙龈炎、牙周炎患者牙周临床指标均得到明显改善,牙周组织炎症状态的改善也伴随着龈沟液内NO水平的下降,而有学者发现,在牙周疾病静止期、痊愈期,龈沟液内NO变化显著[18],因此,通过检测牙龈炎和牙周炎患者龈沟液内NO水平变化,不仅能够判断治疗效果、预测疾病进展,还可评估牙周疾病活动期,为治疗方案的调整提供重要参考。

参 考 文 献

[1] Leppilahti J M, Hernández‐Ríos P A, Gamonal J A, et al. Matrix metalloproteinases and myeloperoxidase in gingival crevicular fluid provide site‐specific diagnostic value for chronic periodontitis[J]. J Clin Periodontol, 2014, 41(4): 348-356.

[2] Belibasakis G N, ?ztürk V ?, Emingil G, et al. Soluble triggering receptor expressed on myeloid cells 1 (sTREM-1) in gingival crevicular fluid: association with clinical and microbiologic parameters[J]. J Periodontol, 2014, 85(1): 204-210.

[3] 戚慧. 一氧化氮与牙周病相关性的实验研究[D]. 昆明:昆明医学院, 2010.

[4] Pourabbas R, Kashefimehr A, Rahmanpour N, et al. Effects of photodynamic therapy on clinical and gingival crevicular fluid inflammatory biomarkers in chronic periodontitis: a split-mouth randomized clinical trial[J]. J Periodontol, 2014, 85(9): 1222-1229.

[5] Leppilahti J M, Kallio M A, Tervahartiala T, et al. Gingival crevicular fluid matrix metalloproteinase-8 levels predict treatment outcome among smokers with chronic periodontitis[J]. J Periodontol, 2014, 85(2): 250-260.

[6] 王艳芝, 李纾. 一氧化氮等气体信号分子与牙周疾病[J]. 国际口腔医学杂志, 2014, 41(2): 204-208.

[7] Baltac?o?lu E, Kehribar M A, Yuva P, et al. Total oxidant status and bone resorption biomarkers in serum and gingival crevicular fluid of patients with periodontitis[J]. J Periodontol, 2014, 85(2): 317-326.

[8] 德特恩维拉. 重度牙周炎治疗临床指南[M]. 北京:人民军医出版社, 2008.

[9] Papathanasiou E, Teles F, Griffin T, et al. Gingival crevicular fluid levels of interferon‐γ, but not interleukin‐4 or‐33 or thymic stromal lymphopoietin, are increased in inflamed sites in patients with periodontal disease[J]. J Periodontol Res, 2014, 49(1): 55-61.

[10] Aurer A, Aleksi? J, Ivi?kardum M, et al. Nitric oxide synthesis is decreased in periodontitis.[J]. J Clin Periodontol, 2001, 28(6):565-568.

[11] 缪羽, 王丽娟, 谢学英. 不同材料全冠修复体对牙周组织及龈沟液成分影响的研究[J]. 中华老年口腔医学杂志, 2015, 13(1): 51-53.

[12] Gul S S, Douglas C W I, Griffiths G S, et al. A pilot study of active enzyme levels in gingival crevicular fluid of patients with chronic periodontal disease[J]. J Clin Periodontol, 2016, 43(8): 629-636.

[13] Kinney J S, Morelli T, Oh M, et al. Crevicular fluid biomarkers and periodontal disease progression[J]. J Clin Periodontol, 2014, 41(2): 113-120.

[14] 沈妍欣, 郭淑娟, 吳亚菲. 慢性牙周炎的氧化应激及抗氧化治疗研究进展[J]. 中华口腔医学杂志, 2016, 51(7): 442-446.

[15] Oh H, Hirano J, Takai H, et al. Effects of initial periodontal therapy on interleukin-1β level in gingival crevicular fluid and clinical periodontal parameters[J]. J Oral Sci, 2015, 57(2): 67-71.

[16] A?ral? ? B, Kuru B E, Yarat A, et al. Evaluation of gingival crevicular fluid transforming growth factor?β1 level after treatment of intrabony periodontal defects with enamel matrix derivatives and autogenous bone graft: A randomized controlled clinical trial[J]. Niger J Clin Pract, 2016, 19(4): 535-543.

[17] Singhal S, Pradeep A R, Kanoriya D, et al. Human soluble receptor for advanced glycation end products and tumor necrosis factor-α as gingival crevicular fluid and serum markers of inflammation in chronic periodontitis and type 2 diabetes[J]. J Oral Sci, 2016, 58(4): 547-553.

[18] Leppilahti J M, Sorsa T, Kallio M A, et al. The utility of gingival crevicular fluid matrix metalloproteinase-8 response patterns in prediction of site-level clinical treatment outcome[J]. J Periodontol, 2015, 86(6): 777-787.

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