新疆桑白皮提取物对主要致龋细菌生物膜作用的实验研究  

Experimental study of effect of Xinjiang Mori Cortexextract on maincariogenic bacteriabiofilm

作　　者：吴龙[1,2] 阿尔曼·阿卜力孜 玛依奴尔·阿塔吾拉 赵今 WU Long;Aerman Abulizi;Mayinuer Atawula;ZHAO Jin(Department of Cariology and Endodontics,the First Affiliated Hospital of Xinjiang Medical University/Affiliated Stomatology Hospital;Institute of Stomatology of Xinjiang Uyghur Autonomous Region,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院(附属口腔医院)牙体牙髓病科 [2]新疆维吾尔自治区口腔医学研究所,乌鲁木齐830054

出　　处：《新疆医科大学学报》2024年第4期447-454,460,共9页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金(2022D01C751);新疆维吾尔自治区“天山英才”科技领军创新人才项目(2023TSYCLJ0032);新疆医科大学第一附属医院“青年科研起航”专项基金(2022YFY-QKQN-40)。

摘　　要：目的探讨新疆桑白皮提取物对主要致龋细菌在生物膜状态下产酸、产糖等致龋毒力因子及生物膜活性的影响。方法制备变异链球菌、血链球菌和黏性放线菌3种单菌种菌悬液,分为阴性对照组[不添加新疆桑白皮提取物或氯己定(CHX)];阳性对照组(添加0.05%CHX);实验组(添加不同浓度新疆桑白皮提取物),测定新疆桑白皮提取物对3种致龋细菌生物膜状态下产酸能力的影响,通过测定24 h内不同时间节点pH值制作pH曲线。采用蒽酮硫酸法测定新疆桑白皮提取物对水溶性及水不溶性胞外多糖合成水平的影响。分别采用乳酸脱氢酶(Lactate dehydrogenase,LDH)活性试验和钙离子(Ca^(2+))泄漏实验,测定新疆桑白皮提取物对3种致龋菌生物膜上清液中LDH含量和Ca^(2+)浓度的影响;采用活死菌染色实验,测定新疆桑白皮提取物对3种致龋菌生物膜活性的影响。结果与阴性对照组比较,实验组(除1/4MBIC_(50)实验组)和CHX组生物膜ΔpH值均减小,3种致龋细菌单菌种生物膜产IEPS、SEPS能力降低,上清液中LDH含量和Ca^(2+)浓度升高,生物膜细菌总量下降,死细菌数量增大,活/死细菌比值百分比下降,差异有统计学意义(P<0.05)。与CHX组比较,实验组3种致龋细菌生物膜ΔpH值均增大,生物膜产IEPS、SEPS能力升高,上清液中LDH含量和Ca^(2+)浓度降低,差异有统计学意义(P<0.05)。结论新疆桑白皮对3种致龋细菌生物膜状态下产酸、产糖能力具有一定抑制作用,能够影响破坏3种致龋细菌生物膜的活性。Objective To explore the effects of Xinjiang Mori Cortex extract on cariogenic virulence factors,such as acid and sugar production of major cariogenic bacteria in biofilm state,as well as biofilm activity.Methods Prepared suspensions of 3 single bacterial strains,namely Streptococcus mutans,streptococcus sanguineum,and streptococcus mucilaginosus,and divided them into a negative control group[without adding Xinjiang Mori Cortex extractor chlorhexidine(CHX)]and positive control group(with 0.05%CHX added).The experimental group(with different concentrations of Xinjiang Mori Cortex extract added)measured the effect of Xinjiang Mori Cortex extract on the acid production ability of 3 cariogenic bacteria under biofilm conditions.pH curves were created by measuring the pH values at different time points within 24 h.The anthrone sulfuric acid method was used to determine the effect of Xinjiang Mori Cortex extract on the synthesis levels of soluble and insoluble extracellular polysaccharides.Using lactate dehydrogenase(LDH)activity assay and calcium ion(Ca^(2+))leakage assay,the effects of Xinjiang Mori Cortex extract on LDH content and Ca^(2+)concentration in the supernatant of 3 cariogenic bacterial biofilms were determined.Using live and dead bacteria staining experiments,the effect of Xinjiang Mori Cortex extract on the biofilm activity of 3 cariogenic bacteria was determined.Results Compared with negative control group,the experimental group(excluding 1/4MBIC_(50) experimental group)and CHX group had biofilms,ΔpH values were decreased,and the ability of 3cariogenic bacteria to produce IEPS and SEPS in biofilms were decreased.The LDH content and Ca^(2+)concentration in the supernatant was increased,and the total number of biofilm bacteria was decreased.The number of dead bacteria wasincreased,and the percentage of live/dead bacteria was decreased,with statistical significance(P<0.05).Compared withCHX group,the experimental group had3 types of cariogenic bacterial biofilms,ΔpH values were increased,and the ability of

关 键 词：龋病 桑白皮 致龋菌 生物膜 

分 类 号：R780.1[医药卫生—口腔医学]