不同低聚糖对变异链球菌、牙龈卟啉单胞菌和具核梭杆菌作用的体外研究

杨云; 姜国庆; 鲁绯; 李晶

doi:10.13386/j.issn1002-0306.2022060023

不同低聚糖对变异链球菌、牙龈卟啉单胞菌和具核梭杆菌作用的体外研究

doi: 10.13386/j.issn1002-0306.2022060023

杨云^{1, 2,},
姜国庆^{2, 3},
鲁绯^{2, 3},
李晶^{2, 3, ,}

1.
中国农业大学食品科学与营养工程学院，北京 100083
2.
北京市营养源研究所，北京 100069
3.
北京市系统营养工程技术研究中心，北京 100069

基金项目: 丰台区科技新星计划项目（Kjxx202004）；北京市科协青年人才托举工程。

详细信息

作者简介:
杨云（1995−），女，硕士研究生，研究方向：天然产物与功能食品，E-mail：yunyangy@163.com

通讯作者:
李晶（1987−），女，博士，副研究员，研究方向：食品生物技术，E-mail：lijing_8715@sina.com

中图分类号: TS202.1
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出版历程
- 收稿日期: 2022-06-10
- 网络出版日期: 2023-01-12
- 刊出日期: 2023-03-01

Effect of Different Oligosaccharides on Streptococcus mutans, Porphyromonas gingivalis and Clostridium nucleatus in Vitro

YANG Yun^{1, 2
,},
JIANG Guoqing^{2, 3},
LU Fei^{2, 3},
LI Jing^{2, 3
, ,}

1.
College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China
2.
Beijing Institute of Nutritional Resources, Beijing 100069, China
3.
Beijing System Nutrition Engineering Technology Research Center, Beijing 100069, China

摘要

摘要: 为更好地了解功能性低聚糖对口腔微生物的影响，本文采用体外实验方法，研究不同低聚糖对变异链球菌、牙龈卟啉单胞菌和具核梭杆菌生长、产酸、粘附性和生物被膜形成量等的影响。胰蛋白胨大豆肉汤（Trypticase Soy Broth，TSB）培养基中加入不同浓度的低聚半乳糖、低聚木糖、低聚果糖和低聚异麦芽糖制备培养基培养变异链球菌、牙龈卟啉单胞菌和具核梭杆菌，测定细菌在培养基中48 h生长过程中的OD值、pH、黏附能力及生物被膜形成量等指标。比较分析不同低聚糖对细菌发酵前后生长、产酸及黏附能力等特性的影响。结果表明：四种低聚糖对三种病原菌均有不同程度的抑制作用。其中，低聚木糖对三种病原菌的生长抑制效果最强；浓度为0.5%的低聚木糖和低聚半乳糖即能明显抑制三种菌液的产酸能力；相比糖醇和其它低聚糖，低聚木糖浓度≥5%时，对三种病原菌的抑制率均大于60%；7%浓度的四种低聚糖对三种病原菌的黏附率均<50%；低聚半乳糖和低聚木糖浓度>5%时，对三种病原菌生物被膜的形成具有较好的抑制作用。与山梨糖醇相比，复配糖（低聚木糖、木糖醇、低聚半乳糖）对三种病原菌的生长抑制作用更为明显。
- 低聚糖 /
- 变异链球菌 /
- 牙龈卟啉单胞菌 /
- 具核梭杆菌 /
- 产酸 /
- 黏附性 /
- 生物膜
Abstract: To provide a theoretical basis for the effects of functional oligosaccharides on the oral microorganisms, the efficacy of several oligosaccharides on the growth, acid production, adherence and biofilm formation of three cariogenic bacteria including Streptococcus mutans, Porphyromonas gingivalis, Fusobacterium nucleatum were investigated. The three cariogenic bacteria were cultured in TSB medium with xylooligosaccharides, galactooligosaccharides, fructooligosaccharide and isomaltooligosaccharide at the different concentrations, and the absorbance, pH, adhesiveness as well as biofilm formation inhibitory activity were measured for 48 h. The effects of different oligosaccharides on bacterial growth, acid production and adhesion were compared and analyzed. The results showed that four oligosaccharides had inhibitory effects on three kinds of cariogenic bacteria, and xylooligosaccharide had the highest inhibitory effect. Xylo-oligosaccharides and galactooligosaccharides at 0.5% concentration could significantly inhibit the acid production of the three cariogenic bacteria. Compared with xylitol and other oligosaccharides, the inhibition rate of xylo-oligosaccharide on the three pathogens was 60% higher when the xylo-oligosaccharide concentration was more than 5%. The adhesion rates of the four oligosaccharides at 7% concentration to the three cariogenic bacteria were all less than 50%. When the concentration of galactooligosaccharides and xylo-oligosaccharide was greater than 5%, the biofilm formation of the three cariogenic bacteria was inhibited well. By comparison with sorbitol, the combination use of xylo-oligosaccharide, xylitol, galactooligosaccharides had more obvious inhibition on the growth of the three pathogenic bacteria.
- oligosaccharide /
- Streptococcus mutans /
- Porphyromonas gingivalis /
- Fusobacterium nucleatum /
- acid production /
- adhesion /
- biofilm formation

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图 1 培养时间对菌液吸光度值的影响

Figure 1. Influence of culture time on absorbance of bacterial solution

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图 2 培养时间对菌液pH的影响

Figure 2. Effect of culture time on the pH of bacterial solution

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图 3 不同浓度低聚糖对病原菌pH变化程度的影响

Figure 3. Effects of different concentrations of oligosaccharides on acidification of bacterial solution

注：小写字母为组内方差分析，不同字母表明具有显著性差异（P<0.05），图5、图6同。

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图 4 低聚糖对三种病原菌生物被膜形成量的抑制作用

Figure 4. Inhibiting effect of oligosaccharides on biofilm formation of three pathogenic bacteria

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图 5 不同细菌在复合糖和山梨糖醇中培养OD值的变化

Figure 5. Changes of OD values of different bacteria cultured in complex sugar and sorbitol

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图 6 不同细菌在复合糖和山梨糖醇中pH的变化

Figure 6. Changes of pH value of different bacteria cultured in complex sugar and sorbitol

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表 1 低聚糖对口腔病原菌的抑制率（%）

Table 1. Inhibition rate of oligosaccharide on oral pathogenic bacteria (%)

糖种类	具核梭杆菌			牙龈卟啉单胞菌			变异链球菌
糖种类	2%	5%	7%	2%	5%	7%	2%	5%	7%
山梨糖醇	14.85±1.32^bC	34.75±0.05^aC	37.23±0.07^aC	16.31±2.67^bC	18.22±0.88^bCD	22.61±1.27^aE	28.81±3.98^abC	35.28±2.99^abC	41.06±1.81^aB
木糖醇	24.66±3.08^aB	22.87±0.85^aD	23.96±0.99^aD	27.06±3.19^abB	32.75±0.14^aB	28.67±1.37^abD	30.72±1.53^abC	41.95±8.58^aC	41.06±1.81^aB
低聚半乳糖	19.81±2.35^bBC	20.59±2.97^bD	38.92±0.36^aC	25.78±3.21^bcB	33.01±3.19^bB	52.88±1.27^aB	37.21±1.15^abBC	36.94±1.98^abC	43.24±0.64^aB
低聚木糖	48.51±0.57^cA	68.61±0.24^bA	89.51±2.18^aA	39.96±4.29^bA	85.20±3.44^aA	88.10±0.88^aA	59.39±3.42^cA	71.10±2.39^bA	88.38±1.0^aA
低聚果糖	18.45±4.06^bBC	50.42±0.74^aB	55.72±0.98^aB	1.86±1.27^cD	16.97±2.19^bCD	34.43±1.63^aC	28.99±2.03^cC	54.78±0.75^aB	44.52±0.91^bB
低聚异麦芽糖	5.23±1.53^aD	8.94±1.49^aE	10.23±5.54^aE	13.09±1.98^aC	15.03±1.06^aD	15.51±2.88^aF	13.56±2.34^bD	19.26±0.78^aD	21.36±2.07^aC
注：小写字母为组内方差分析，大写字母为组间方差分析，不同字母表明具有显著性差异（P<0.05），表2同。

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表 2 不同浓度低聚糖对三种病原菌黏附性的影响（%）

Table 2. Effects of different concentrations of oligosaccharides on adhesion of three pathogenic bacteria (%)

糖种类	具核梭杆菌			牙龈卟啉单胞菌			变异链球菌
糖种类	2%	5%	7%	2%	5%	7%	2%	5%	7%
山梨糖醇	68.55±0.56^aA	55.53±0.22^bAB	42.49±0.30^cA	46.96±0.09^bC	59.73±0.04^aA	46.68±0.22^bA	56.76±0.37^bA	66.32±0.14^aA	48.90±0.09^cA
木糖醇	49.90±0.14^bD	55.66±0.60^aAB	34.89±0.32^cC	35.34±0.45^bD	49.44±0.12^aC	33.46±0.69^cD	41.64±0.59^bCD	51.68±0.21^aD	24.77±0.97^cD
低聚半乳糖	41.41±0.04^aE	44.50±1.41^aBC	36.41±0.39^cB	41.47±0.04^cC	49.31±0.09^aC	41.00±0.78^bB	56.69±0.06^aA	56.09±0.61^aC	39.63±1.62^bB
低聚木糖	50.90±0.62^aC	42.94±1.47^bC	33.80±0.01^cC	52.11±1.57^aB	36.86±0.17^bD	33.57±0.33^cD	53.94±0.09^bB	50.77±2.16^aD	36.28±0.73^bBC
低聚果糖	51.36±0.64^aCD	31.43±9.11^aD	37.04±0.40^aB	31.28±0.33^cE	50.62±1.53^aC	40.19±1.87^bBC	46.15±4.15^bC	62.84±1.42^aB	42.01±0.80^bB
低聚异麦芽糖	53.25±0.30^bB	65.08±1. 13^aA	34.55±0.71^cC	53.46±0.13^aA	54.94±3.49^aB	38.73±1.41^bC	40.49±0.13^aD	41.16±0.45^aE	31.90±5.12^aC

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参考文献(32)

[1]	单贤文, 梁卫平, 何月华. 口腔健康状况与慢性胃炎的相关性[J]. 医疗装备,2021,34(14):101−102. [SHAN X W, LIANG W P, HE Y H. Relationship between oral health status and chronic gastritis[J]. Medical Equipment,2021,34(14):101−102. doi: 10.3969/j.issn.1002-2376.2021.14.052
[2]	KILIAN M, CHAPPLE I L C, HANNIG M, et al. The oral microbiome-an update for oral healthcare professionals[J]. British Dental Journal,2016,221(10):657−666. doi: 10.1038/sj.bdj.2016.865
[3]	邓盟, 齐霞, 徐欣, 等. 人体口腔微生物宏基因组的研究进展[J]. 华西口腔医学杂志,2013,31(1):107−111. [DENG M, QI X, XU X, et al. Latest progress in unveiling the oral microbial metagenome[J]. West China Journal of Stomatology,2013,31(1):107−111. doi: 10.7518/hxkq.2013.01.027
[4]	SAMPAIO-MAIA B, CALDAS I M, PERIRA M L, et al. The oral microbiome in health and its implication in oral and systemic diseases[J]. Advances in Applied Microbiology,2016:171−210.
[5]	HAN S J, JEONG S Y, NAM Y J, et al. Xylitol inhibits inflammatory cytokine expression induced by lipopolysaccharide from Porphyromonas gingivalis[J]. Clinical and Diagnostic Laboratory Immunology,2005,12(11):1285−1291.
[6]	周琳, 蔡文静, 杨慧, 等. MTAN用于抑制血链球菌、具核梭杆菌、牙龈卟啉单胞菌混合菌的体外研究[J]. 安徽医科大学学报,2021,56(8):1328−1331. [ZHOU L, CAI W J, YANG H, et al. Vitro study of MTAN to inhibit the mixed bacteria solution of S. anguinis, F. nucleatum and P. gingivalis[J]. Acta Universitatis Medicinalis Anhui,2021,56(8):1328−1331. doi: 10.19405/j.cnki.issn1000-1492.2021.08.032
[7]	SOJAR H T, SMITH D F. Porphyromonas gingivalis fimbriae carbohydrate specificity assessment by glycomics[J]. Fems Immunology and Medical Microbiology,2012,66(1):83−87. doi: 10.1111/j.1574-695X.2012.00989.x
[8]	蒋端, 申道南. 具核梭杆菌影响动脉粥样硬化的分子机制[J]. 口腔医学研究,2022,38(4):308−311. [JIANG D, SHEN D N. Research on molecular mechanism of effect of Fusobacterium nucleatum on atherosclerosis[J]. Journal of Oral Science Research,2022,38(4):308−311. doi: 10.13701/j.cnki.kqyxyj.2022.04.003
[9]	张程, 孙红英. 口腔微生物组与全身疾病的相关性[J]. 生理科学进展,2021,52(2):128−132. [ZHANG C, SUN H Y. Correlation between oral microbiome and systemic diseases[J]. Progress in Physiological Sciences,2021,52(2):128−132. doi: 10.3969/j.issn.0559-7765.2021.02.011
[10]	张佳丽, 姚军, 诸葛继娜, 等. 赤藓糖醇对牙周致病菌抗菌性能的影响[J]. 上海口腔医学,2019,28(4):362−367. [ZHANG J L, YAO J, ZHUGE J N, et al. Antibacterial activity of erythritol on periodontal pathogen[J]. Shanghai Journal of Stomatology,2019,28(4):362−367. doi: 10.19439/j.sjos.2019.04.005
[11]	李秀娟, 韩雪松, 钟滨, 等. 糖醇与龋病关系的研究进展[J]. 口腔医学,2013,33(10):713−714. [LI X J, HAN X S, ZHONG B, et al. Research progress on the relationship between sugar alcohols and caries[J]. Stomatology,2013,33(10):713−714. doi: 10.13591/j.cnki.kqyx.2013.10.016
[12]	黄晨辉, 王伟健, 杨圣辉. 异麦芽酮糖醇对变形链球菌生长、黏附和产酸影响的研究[J]. 现代口腔医学杂志,2008(4):362−364. [HUANG C H, WANG W J, YANG S H. Effect of isomalt on amount, adhesion, and pH value of Streptococcus mutans[J]. Journal of Modern Stomatology,2008(4):362−364. doi: 10.3969/j.issn.1003-7632.2008.04.008
[13]	董占能, 邢思敏. 一类新型保健食品——低聚糖[J]. 昆明理工大学学报,1998,6:68−72. [DONG Z N, XING S M. A norel healthful food—oligosaccharide[J]. Journal of Kunming University of Science and Technology,1998,6:68−72.
[14]	邹月, 黄金凤, 魏琴. 功能性低聚糖的研究进展及应用现状[J]. 中国调味品,2021,46(2):180−185, 195. [ZOU Y, HUANG J F, WEI Q. Research progress and application status of functional oligosaccharides[J]. China Condiment,2021,46(2):180−185, 195. doi: 10.3969/j.issn.1000-9973.2021.02.039
[15]	肖敏. 功能性低聚糖及其生产应用[J]. 生物产业技术,2018,6:29−34. [XIAO M. Functional oligosaccharides and their production and application[J]. Bioindustry Technology,2018,6:29−34.
[16]	TSUKATANI T, SAKATA F, KURODA R. A rapid and simple measurement method for biofilm formation inhibitory activity using 96-pin microtiter plate lids[J]. World Journal of Microbiology & Biotechnology,2020,36(12):189.
[17]	GAIKWAD S S, GHEWARE A, KAMATAGI L, et al. Dental caries and its relationship to malocclusion in permanent dentition among 12-15 year old school going children[J]. Journal of International Oral Health Jioh,2014,6(5):27−30.
[18]	PINNA R, USAI P, FILIGHEDDU E, et al. The role of adhesive materials and oral biofilm in the failure of adhesive resin restorations[J]. American Journal of Dentistry,2017,30(5):285−292.
[19]	ROSIER B T, DE JAGER M, ZAUAR E, et al. Historical and contemporary hypotheses on the development of oral diseases: are we there yet?[J]. Frontiers in Cellular and Infection Microbiology,2014,4(4):92.
[20]	SAMPAIOMAIA S M. Acquisition and maturation of oral microbiome throughout childhood: An update[J]. Dental Research Journal,2014,11(3):291−301.
[21]	TAHMASSEBI J F, DUGGAL M S. The effect of different methods of drinking on the pH of dental plaque in vivo[J]. International Journal of Paediatric Dentistry,2010,7(4):249−254.
[22]	AL D A, ATWA R Y A, MOSTAFA M. Effect of honey in preventing gingivitis and dental caries in patients undergoing orthodontic treatment[J]. Saudi Dental Journal,2014,26(3):108−114.
[23]	KARTHIKEYAN R, AMAECHI B T, RAWLS H R, et al. Antimicrobial activity of nanoemulsion on cariogenic Streptococcus mutans[J]. Archives of Oral Biology,2011,56(5):437−445.
[24]	KRZYSCIAK W, JURCZAK A, KOSCIELNIAK D, et al. The virulence of Streptococcus mutans and the ability to form biofilms[J]. European Journal of Clinical Microbiology & Infectious Diseases,2014,33(4):499−515.
[25]	DOMON H, HIYOSHI T, MAEKAWA T, et al. Antibacterial activity of hinokitiol against both antibiotic-resistant and -susceptible pathogenic bacteria that predominate in the oral cavity and upper airways[J]. Microbiology and Immunology,2019,63(6):213−222. doi: 10.1111/1348-0421.12688
[26]	林丹乐, 谢永婷, 王雪梅. 低聚木糖对变异链球菌生长和产酸影响的体外研究[J]. 中国实用口腔科杂志,2019,12(8):480−485. [LIN D L, XIE Y T, WANG X M. An in vitro study on the growth and acid production of Streptococcus mutans affected by xylooligosaccharides[J]. Chinese Journal of Practical Stomatology,2019,12(8):480−485. doi: 10.19538/j.kq.2019.08.007
[27]	李维丹. 赤藓糖醇对牙龈卟啉单胞菌的生长、黏附及胰酶样蛋白酶活性的影响分析[D]. 福州: 福建医科大学, 2016. LI W D. Analysis of the effect of erythritol on the growth, adhesion and activity of trypsin-like protease of Porphyromonas gingivalis[D]. Fuzhou: Fujian Medical University, 2016
[28]	MARSH P D. Microbiology of dental plaque biofilms and their role in oral health and caries[J]. Dental Clinics of North America,2010,54(3):441−454. doi: 10.1016/j.cden.2010.03.002
[29]	TAMURA S, YONEZAWA H, MOTEGI M, et al. Inhibiting effects of Streptococcus salivarius on competence-stimulating peptide-dependent biofilm formation by Streptococcus mutans[J]. Oral Microbiology and Immunology,2009,24(2):152−161. doi: 10.1111/j.1399-302X.2008.00489.x
[30]	李懿洋, 周学东. 具核梭杆菌与口腔常见微生物粘附作用的研究进展[J]. 口腔医学研究,2021,37(4):284−287. [LI Y Y, ZHOU X D. Overviews of research on coadherence between Fusobacterium nucleatum and common oral microorganisms[J]. Journal of Oral Science Research,2021,37(4):284−287. doi: 10.13701/j.cnki.kqyxyj.2021.04.002
[31]	HARIRIAN H, AnDRUKHOV O, BERTL K, et al. Microbial analysis of subgingival plaque samples compared to that of whole saliva in patients with periodontitis[J]. Journal of Periodontology,2014,85(6):819−828.
[32]	郭宏磊, 王永兰, 靳趁心. 木糖醇对黏放线菌生长、产酸和黏附影响的实验研究[J]. 全科口腔医学电子杂志,2015,2(10):102−103. [GUO H L, WANG Y X, JIN C X. Experimental study on the effects of xylitol on Actinomycetes growth, acid production and adhesion[J]. General Journal of Stomatology,2015,2(10):102−103.