胃癌前病变患者口腔细菌多样性分析

Oral microbial diversity in individuals with gastric precancerous lesions

目的分析胃癌前病变患者口腔中唾液和牙菌斑样本的细菌多样性,以期获得口腔菌群特征与胃癌的相关性的科学证据。方法选取约定进行上消化道内窥镜检查的97名应试者。按病理诊断结果,将受试者分为2组,对照组无任何病理变化;病变组患有胃癌前病变(包括慢性萎缩性胃炎、肠上皮化生或异型增生)。提取唾液和牙菌斑样本DNA,进行巢式PCR变性梯度凝胶电泳,采集DGGE图像,BioNumerics软件分析图像,SPSS17.0统计软件分析微生物学指标和细菌多样性之间的相关性和相似性。结果胃癌前病变患者的龈下菌斑样本的DGGE条带数(35.6)显著高于对照组(33.2)(P<0.05),但2组间唾液样本的DGGE条带数量,未见显著性差异。所有受试者的6个不同牙位点龈下菌斑之间的相似性达80.7%,而唾液和龈下菌斑集合2种不同来源样本的菌群相似性只有78.0%。病变组唾液样本和龈下菌斑样本的相似程度(76.5%)略低于对照组(78.6%);病变组不同牙位龈下菌斑样本之间的相似程度比对照组略高。结论胃癌前病变患者口腔中龈下菌斑的细菌多样性略高于对照组,不同牙位龈下菌斑细菌多样性的相似程度高于唾液和龈下菌斑。

Objective To analyze the microbial populations obtained from both saliva and subgingival plaque samples from individuals with gastric precancerous lesions and to investigate the association between oral flora and gastric precancerous lesions. Methods A total of 97 individuals who were scheduled for an upper endoscopy procedure at the gastrointestinal clinics were included. All individuals aged over 30 years old and had no history of esophageal cancer, gastric cancer,gastric varices, or portal gastropathy. Based on biopsy diagnosis, these individuals were divided into two groups, individuals with gastric precancerous lesion （chronic atrophic gastritis, intestinal metaplasia or dysplasia）, individuals with no lesions. Bacterial samples were taken from all 97 subjects at the time of dental examination. Bacterial genomie DNA of the plaque or saliva was isolated. Nested PCR was performed with use of the GeneAmp PCR SystemTM 9700. PCR products and species-specific DGGE reference markers were directly loaded in each lane, and electrophoresis was performed. DGGE profile images were digitally captured and recorded and then analyzed using the BioNumerics Software. Results Subjects with gastric precancerous lesions had more DGGE bands （35.6） in pooled plaque sample, compared with healthy population （33. 2） ,while there was no significant difference in saliva sample. The similarity of diversity between saliva and pooled plaque was 78.0% ,lower than that among 6 subgingival plaques（80. 7% ）. The similarity of diversity between saliva and pooled plaque of the subjects with gastric precancerous lesions was lower compared with the control group. The similarity among 6 subgingival plaques of the individuals with gastric precancerous lesion was higher than that of the control. Conclusion Subjects with gastric precancerous lesions had more microbial diversity in pooled subgingival plaque compared with healthy population. The similarity between subgingival plaque samples was higher than that between saliva and subgingival plaque samples.