高脂高果糖饮食与维生素D缺乏对小鼠牙龈菌群影响的实验研究

Experimental research in the effect of high fat high fructopyranose and Vitamin D receptor-knock-out on subgingival bacterial diversity in mice

目的检测在高脂高果糖饮食和维生素D缺乏条件下小鼠龈下菌群变化,探讨其导致牙周病变的可能机制。方法正常型和维生素D受体敲出的野生型C57BL/6J小鼠各20只,各自随机分成正常饮食组和高脂高果糖饮食组,每组10只。喂养10周后检测下颌第2磨牙牙周炎临床指标,提取龈沟液,采用PCR-变形梯度凝胶电泳技术(PCR-DGGE)和DNA测序等方法分析龈沟液的菌群差异性。结果与正常型高脂高果糖组比较,维生素D受体敲出型高脂高果糖组在牙龈指数、牙周袋深度、牙松动度方面差异有统计学意义(P<0.05);正常型高脂高果糖组、维生素D敲出型组与正常型正常饮食组比较,小鼠龈沟液菌群分布差异有统计学意义(P<0.05);测序结果显示正常型高脂高果糖组和维生素D受体敲出型高脂高果糖组小鼠的优势菌群不完全相同。结论通过成功建立所需牙周炎动物实验模型,初步证明维生素D缺乏对高脂高果糖所致牙周炎的发展可能存在正协同作用。

Objective To detect the oral bacterial diversity in mice under high fat and high fructopyranose diet or/and Vitamin D receptor-knock-out conditions,in order to study the mechanism between the two conditions and periodontitis.MethodsC57BL/6Jwild type mice including 20 normal ones and 20 Vitamin D receptor-knock-out ones,were divided into normal diet and high fat and high fructopyranose diet groups respectively,10 cases in each group.After feeding 10 weeks,collecting gingival crevicular fluid from second mandible molar,analysising the bacterial diversity of the gingival crevicular fluid by using Polymerase chain reaction denaturing gradient gel electrophoresis（PCR-DGGE）method.Collecting specific bands for DNA sequence assay.ResultsComparing with normal mice feeding with high fat and high fructopyranose diet,the gingival index,periodontal pocket and tooth mobility of Vitamin D receptor-knock-out mice feeding with high fat and high fructopyranose diet had obvious statistical difference（P〈0.05）;the distribution of the gingival bacterial diversity between normal mice with normal food and other 3groups had statistical difference（P〈0.05）.The results of sequence assay showed that the dominan bacteria between normal mice with high fat and high fructopyranose diet and Vitamin D receptor-knock-out mice with high fat and high fructopyranose diet was not the same.Conclusion It has successfully built a animal model for periodontitis research of high fat and high fructopyranose diet or/and Vitamin D deficient,the periodontitis will get worse under the effect of the two factor.