糖尿病伴慢性牙周炎患者口腔菌群变化及其与龈沟液炎症因子的相关性研究

Study on the Changes of Oral Flora and Its Correlation with Gingival Crevicular Fluid Inflammatory Factors in Diabetes Patients with Chronic Periodontitis

目的观察糖尿病伴慢性牙周炎患者口腔菌群变化,并分析其与龈沟液炎症因子的相关性。方法选取保定市第一中心医院2022年6~12月收治的86例糖尿病伴慢性牙周炎患者、88例不伴糖尿病的慢性牙周炎患者分别记为A组和B组,另于同期招募85例健康志愿者记为对照组。三组均采用16S rDNA高通量测序技术检测口腔菌群属水平的相对丰度,采用酶联免疫吸附法检测龈沟液炎症因子水平[包括超敏C反应蛋白(high sensitivity C-reactive protein,hs-CRP)、肿瘤坏死因子-α(tumornecrosisfactor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)、白细胞介素-6(interleukin-6,IL-6)和白细胞介素-4(interleukin-4,IL-4)]。比较三组口腔菌群属水平相对丰度、多样性指数和龈沟液炎症因子水平;采用Pearson法分析A组口腔菌群属水平相对丰度与龈沟液炎症因子水平的相关性。结果A组乳杆菌属、双歧杆菌属、酵母菌属和肠球菌属水平相对丰度、龈沟液IL-10水平均低于其余二组(t=4.657~37.617,均P<0.05),B组均低于对照组,差异均具有统计学意义(t=7.378~29.378,均P<0.05);A组卟啉单胞菌属、韦荣球菌属、普氏菌属、奈瑟菌属和密螺旋体属水平相对丰度、龈沟液hs-CRP,TNF-α,IL-6和IL-4水平均高于其余二组(t=2.945~39.822,均P<0.05),B组均高于对照组,差异具有统计学意义(t=2.949~26.347,均P<0.05)。A组和B组Shahnon指数(5.44±0.48,5.35±0.36)均高于对照组(5.24±0.18),差异有统计学意义(t=3.599,2.528;P=0.000,0.012)。A组乳杆菌属、双歧杆菌属、酵母菌属和肠球菌属水平相对丰度与龈沟液IL-10水平均呈正相关(r=0.728~0.795,均P<0.05),与龈沟液hs-CRP,TNF-α,IL-6和IL-4水平均呈负相关(r=-0.867~-0.731,均P<0.05);A组卟啉单胞菌属、韦荣球菌属、普氏菌属、奈瑟菌属和密螺旋体属水平相对丰度与龈沟液IL-10水平均呈负相关(r=-0.854~-0.730,均P<0.05),与龈沟液hs-CRP,TNF-α,IL-6和IL-4水平均呈正相关(r=0.705~0.892,均P<0.05)。结论糖尿病伴慢性牙周炎患者口腔致病菌取代有益菌成为优势菌造成口腔菌群失调,龈沟液促炎症因子水平升高而抗炎症因子水平降低,较单纯慢性牙周炎患者改变更为显著,且在伴糖尿病的慢性牙周炎患者中口腔菌群属水平相对丰度与龈沟液炎症因子水平均有密切相关性。

Objective To observe the changes of oral flora in diabetes patients with chronic periodontitis,and analyze the correlation between oral flora and inflammatory factors in gingival crevicular fluid(GCF).Methods 86 diabetes patients with chronic periodontitis and 88 chronic periodontitis patients without diabetes who admitted to the First Central Hospital of Baoding from June 2022 to December 2022 were selected as A group and B group,and 85 healthy volunteers were recruited in the same period as the control group.16S rDNA high throughput sequencing was used to detect the relative abundances of oral flora in the three groups,and the levels of inflammatory factors[including high sensitivity C-reactive protein(hs-CRP),tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),interleukin-6(IL-6)and interleukin-4(IL-4)]in GCF were detected by enzyme linked immunosorbent assay.The relative abundances of oral flora,diversity index and the levels of inflammatory factors in gingival crevicular fluid were compared among the three groups,and the correlation between relative abundances of oral flora and the levels of inflammatory factors in gingival crevicular fluid in group A was analyzed by Pearson method.Results The relative abundances of Lactobacillus,Bifidobacterium,Saccharomyces,Enterococcus and the level of IL-10 in gingival crevicular fluid in group A were lower than those in the other two groups(t=4.657~37.617,all P<0.05),and those in group B were lower than those in the control group(t=7.378~29.378,all P<0.05),the differences were statistically significant,respectively.The relative abundances of Porphyromonas,Veronicus,Prevotella,Neisseria and Treponema and the levels of hs-CRP,TNF-α,IL-6 and IL-4 in gingival crevicular fluid in group A were higher than those in the other two groups(t=2.945~39.822,all P<0.05),and those in group B were higher than those in the control group(t=2.949~26.347,all P<0.05),the differences were statistically significant,respectively.The Shahnon index in group A and group B(5.44±0.48,5.35±0.36)was higher than that in the control group(5.24±0.18),and the difference was statistically significant(t=3.599,2.528,P=0.000,0.012).The relative abundances of Lactobacillus,Bifidobacterium,Saccharomyces and Enterococcus in group A were positively correlated with the level of IL-10 in gingival crevicular fluid(r=0.728~0.795,all P<0.05),which were negatively correlated with the levels of hs-CRP,TNF-α,IL-6 and IL-4 in gingival crevicular fluid(r=-0.867~-0.731,all P<0.05).The relative abundances of Porphyromonas,Veronococcus,Prevotellla,Neisseria and Treponema in group A were negatively correlated with the level of IL-10 in gingival crevicular fluid(r=-0.854~-0.730,all P<0.05),which were positively correlated with the levels of hs-CRP,TNF-α,IL-6 and IL-4 in gingival crevicular fluid(r=0.705~0.892,all P<0.05).Conclusion Oral pathogenic bacteria in diabetes patients with chronic periodontitis replace beneficial bacteria as the dominant bacteria,and causing oral flora disorder,floratheir proportion is high,and the levels of pro-inflammatory factors in gingival crevicular fluid were increased,while the levels of anti-inflammatory factors were decreased,which were more significant than those in patients with simple chronic periodontitis.In the former,the relative abundances of oral flora in chronic periodontitis patients with diabetes mellitus were closely related to the levels of inflammatory factors in gingival crevicular fluid.