中国实用口腔科杂志

中国实用口腔科杂志

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灌饲牙龈卟啉单胞菌对小鼠肠道炎症影响的研究

戴柯1a，王苇如1b，杨钦博1b，刘妮1b，刘洁2，寇育荣1   

  1. 1. 中国医科大学口腔医学院·附属口腔医院a口腔生物学教研室，b牙周病科，辽宁 沈阳 110002；2. 中国医科大学科学实验中心，辽宁 沈阳 110122
  • 出版日期:2019-11-15 发布日期:2020-03-12
  • 基金资助:
    辽宁省自然科学基金项目（20180551232）；沈阳市科技计划项目（F16-102-4-00）；沈阳市中青年科技创新人才支持计划项目（RC170078）

  • Online:2019-11-15 Published:2020-03-12

摘要： 目的　研究灌饲牙龈卟啉单胞菌（Porphyromonas gingivalis，P. gingivalis）对C57bl/6小鼠结肠炎症的诱导作用。方法　将P. gingivalis ATCC33277液体增菌后备用。将15只C57bl/6小鼠适应1周后随机均分为3组，高浓度组灌饲1 × 109 CFU P. gingivalis，低浓度组灌饲1 × 108 CFU P. gingivalis，而对照组则灌饲等量无菌BHI培养液。每只小鼠每天灌饲1次，3周后处死小鼠，采集结肠及脾脏组织，行HE染色观察组织学变化，并采用实时荧光定量PCR（qRT-PCR）检测结肠组织中CD3抗原（CD3 antigen，epsilon polypeptide，CD3）、受体型蛋白酪氨酸磷酸酶C（protein tyrosine phosphatase，receptor type C，B220）、黏附G蛋白偶联受体E1（adhesion G protein-coupled receptor E1，F4/80）、转化生长因子-β（transforming growth factor-β，TGF-β）及干扰素-γ（interferon-gamma，IFN-γ）的表达水平。结果　HE染色显示高浓度组小鼠结肠黏膜下结缔组织中淋巴滤泡增多，且高浓度组小鼠脾指数出现增高的趋势，但差异无统计学意义（P > 0.05）。qRT-PCR结果显示，与对照组及低浓度组相比，高浓度组小鼠的结肠组织中B220及TGF-β的表达水平显著增高（P < 0.05），其余指标表达水平差异无统计学意义（P > 0.05）。结论　P. gingivalis可诱导结肠炎症，从而增加牙周病患者对消化系统疾病的易感性，且其可能与牙周病病情的严重程度相关。

关键词: 肠道, 牙龈卟啉单胞菌, 牙周病, 小鼠

Abstract: Objective　To investigate the effects of orally administered Porphyromonas gingivalis（P. gingivalis）on colonic and systemic inflammation in C57bl/6 mice. Methods　P. gingivalis ATCC33277 were cultured and enriched in brain heart infusion broth（BHI）. Fifteen C57bl/6 mice were randomly divided into three groups. Mice in the high-concentration group were fed with 0.2 mL P. gingivalis（1 × 109 CFU）；those in the low-concentration group were fed with 0.2 mL P. gingivalis（1 × 108 CFU）；mice in the control group were fed with the same amount of BHI. Each mouse was orally administered P. gingivalis once daily. After 3 weeks，the mice were sacrificed and their colons and spleens were collected. The histological changes of colon were observed by HE staining. Real-time quantitative PCR（qRT-PCR）was used to detect the mRNA expression level of some inflammatory factors in the colon，including antigen，epsilon polypeptide（CD3），protein tyrosine phosphatase，receptor type C（B220），adhesion G protein-coupled receptor E1（F4/80），transforming growth factor-β（TGF-β）and interferon-gamma（IFN-γ）. Results　HE staining showed an increase of lymphoid follicles in the colonic submucosal connective tissue of mice in the high-concentration group，and the spleen index of the high-concentration group showed an increasing trend（P > 0.05）. Compared with the low-concentration group and the control group，a significant up-regulation of mRNA expression of TGF-β and B220 was detected in the high-concentration group by qRT-PCR（P < 0.05），while others were no significant（P > 0.05）. Conclusion　P. gingivalis can induce colonic inflammation，thereby may increase the susceptibility of patients with periodontal disease to the digestive system diseases，which can be related to the severity of periodontal disease.

Key words: intestine；Porphyromonas gingivalis, P. gingivalis；periodontal disease；mice