一种由口腔菌群失衡引起的大鼠口臭模型的构建及其评价

doi:10.13701/j.cnki.kqyxyj.2023.08.015

摘要/Abstract

摘要： 目的: 利用口源性口臭(Intra-oral halitosis,IOH)患者唾液整体微生物群定植构建大鼠口臭模型,通过检测大鼠口腔相关微生物、相关代谢和口气值变化对该模型的可行性进行评价。方法: 将健康的6~8周龄雄性Wistar大鼠随机分为两组,模型组(H组)和对照组(C组)。均适应性饲养1周后麻醉下进行口腔菌群采样并检测口气值,使用蘸有生理盐水的棉签机械清洗两组大鼠的牙齿和黏膜。再常规饲养1周后重复清洗并使用0.1%NaoCl冲洗口腔黏膜和牙齿5 min,随后用23 μmol/L缓冲抗坏血酸钠冲洗10 min。使用加入保护液的IOH患者唾液整体微生物群涂抹H组大鼠牙齿和口腔黏膜,每日1次,涂抹后禁食水1 h,后持续给予10%高糖水,6周后麻醉下口腔菌群采样。同时常规饲养C组并采样。采用16SrRNA测序技术检测大鼠口腔微生物变化,使用PICRUSt进行菌群代谢功能预测,利用Halimeter口气检测仪每2周检测大鼠口气值。结果: 与C组相比,H组口腔菌群中卟啉单胞菌属(Porphyromonas)、梭杆菌属(Fusobacterium)、孪生球菌属(Gemella)、纤毛菌属(Leptotrichia)和消化链球菌属(Peptostreptococcus)等与IOH相关的物种比例明显升高。PICRUSt代谢预测分析表明,与C组相比,H组口臭相关代谢途径明显增强。同时口气检测值也出现显著升高。结论: 该造模方法可使IOH相关微生物定植大鼠口腔并发挥其产臭功能,很好的模拟了临床患者口腔菌群和口气特征,可用于未来IOH新的治疗方式和疾病发生发展的研究中。

关键词: 口源性口臭, 口腔菌群, 动物模型

Abstract: Objective: To construct a rat halitosis model using salivary holistic microbiota colonization in patients with intra-oral halitosis (IOH), and to evaluate the feasibility of the model by detecting the changes of oral-associated microorganisms, associated metabolism, and breath values in rats. Methods: The healthy 6-8 week-old male Wistar rats were randomly divided into two groups, the model group (Group H) and the control group (Group C). Oral flora sampling was performed and breath values were measured under anesthesia after 1 week of acclimatization, mechanical cleaning of the teeth and mucosa of both groups of rats using cotton swabs dipped in saline. The cleaning was repeated and the oral mucosa and tooth were rinsed with 0.1% NaoCl for 5 min after 1 week of routine feeding, followed by a 10 min rinse with 23 μM buffered sodium ascorbate. Tooth and oral mucosa of rats in group H were coated using salivary holistic microbiota of IOH patients added with protective solution once daily, with water and food fasting for 1 h after application, followed by continuous administration of 10% high sugar water, and oral flora sampling under anesthesia for 6 weeks. Group C was also routinely reared and sampled. The 16SrRNA sequencing technique was used to detect microbial changes in the rat oral cavity, the PICRUSt was used to predict the metabolic function of the flora, and the Halimeter breath detector was used to test the breath values of the rats every 2 weeks. Results: Compared with group C, the proportion of IOH-associated species such as porphyromonas, fusobacterium, gemella, leptotrichia, and peptostreptococcus in the oral flora of group H was significantly higher. PICRUSt metabolic prediction analysis showed that the metabolic pathways associated with halitosis were significantly enhanced in group H compared with group C. There was also a significant increase in breath test values. Conclusion: This modeling method allows IOH-related microorganisms to colonize the oral cavity of rats and exert their odor-producing function and well simulates the oral flora and breath characteristics of clinical patients, and can be used in future research on new treatment modalities and disease development of IOH.

Key words: intra-oral halitosis, oral flora, animal models

黄志强, 程永波. 一种由口腔菌群失衡引起的大鼠口臭模型的构建及其评价[J]. 口腔医学研究, 2023, 39(8): 745-750.

HUANG Zhiqiang, CHENG Yongbo. Establishment and Evaluation of A Rat Model of Halitosis Caused by Imbalance of Oral Flora[J]. Journal of Oral Science Research, 2023, 39(8): 745-750.

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