慢性牙周炎和2型糖尿病患者肠道柔嫩梭菌类群的定量分析

山东大学学报(医学版) ›› 2013, Vol. 51 ›› Issue (4): 100-104.

慢性牙周炎和2型糖尿病患者肠道柔嫩梭菌类群的定量分析

刘文婷1，胡以俊1，陈丽2，姜广水1

1.山东大学口腔医学院口腔内科教研室，山东省口腔生物医学重点实验室；
2.山东大学齐鲁医院内分泌科，济南 250012

收稿日期:2012-12-09 出版日期:2013-04-10 发布日期:2013-04-10
通讯作者: 姜广水(1958- )，男，博士，教授，主要从事口腔内科疾病的免疫学防治研究。E-mail: gjiang@sdu.edu.cn
作者简介:刘文婷(1986- )，女，硕士研究生，主要从事口腔内科的研究。E-mail: wentinggoodluck@sina.com
基金资助:
国家自然科学基金(30171010，30371323)；山东省科技发展计划(2010GSF10267)

Quantitative analysis of Clostridium leptum subgroup in patients with chronic periodontitis and type 2 diabetes mellitus

LIU Wen-ting1, HU Yi-jun1, CHEN Li2, JIANG Guang-shui1

1. Department of Oral Medicine, School of Stomatology, Shandong University,Shandong Provincial Key Laboratory
of Oral Biomedicine; 2. Department of Endocrinology, Qilu Hospital of Shandong University, Jinan 250012, China

Received:2012-12-09 Online:2013-04-10 Published:2013-04-10

摘要/Abstract

摘要：

目的探讨人肠道柔嫩梭菌(Clostridium leptum，C.leptum) 类群与慢性牙周炎和2型糖尿病的相关关系。方法收集慢性牙周炎患者(CP组)、2型糖尿病患者(DM组)、伴2型糖尿病的慢性牙周炎患者(DM+CP组)及健康对照者(HC组)各20例的粪便标本。根据C.leptum类群16S rRNA序列设计特异性引物，以常规PCR产物经克隆后的质粒DNA为标准品，经梯度稀释后制备标准曲线。待测粪便标本提取细菌基因组DNA，应用实时定量PCR方法对C.leptum类群进行定量分析。结果 4组标本每克湿便中C.leptum类群的16S rRNA基因拷贝数的对数值(log copies/g)：HC组(7.09±0.17)、CP组(6.31±0.58)、DM组(5.79±0.52)、DM+CP组(5.41±0.29)。与HC组相比，其他三组C.leptum类群基因拷贝数均明显减少(P<0.05)。结论 C.leptum类群可能与慢性牙周炎和2型糖尿病的发生、发展相关。

关键词: 慢性牙周炎；2型糖尿病；柔嫩梭菌类群；实时定量聚合酶链式反应

Abstract:

Objective To investigate the relationship between Clostridium leptum (C.leptum) subgroup in the human gut, the periodontal disease and type 2 diabetes mellitus. Methods Fresh fecal samples were obtained from twenty patients with chronic periodontitis (CP group), twenty patients with type 2 diabetes mellitus (DM group), twenty patients with chronic periodontitis complicated by type 2 diabetes ［(DM+CP) group］ and twenty healthy control subjects (HC group). Genus-specific primer of C.leptum subgroup was designed for 16S rRNA-targeted PCR. The standard curve of real-time quantitative polymerase chain reaction (RTQPCR) was generated from a grade-dilution of plasmid DNA which was cloned from PCR products. Bacterial genome DNA extracted from fecal samples was quantified by RTQ-PCR to analyze C.leptum subgroup amounts. Results Logarithm for 16S rRNA gene copies of C.leptum subgroup per gram feces in HC group, CP group, DM group and (DM+CP) group were (7.09±0.17), (6.31±0.58), (5.79±0.52) and (5.41±0.29), respectively. Compared with HC group, the logarithm of the other three groups were dramatically decreased (P＜0.05). Conclusion C.leptum subgroup may be associated with the development and progression of chronic periodontitis and type 2 diabetes mellitus.

Key words: Chronic periodontitis; Type 2 diabetes mellitus; Clostridium leptum subgroup； Real-time quantitative polymerase chain reaction

中图分类号:

R781.4

刘文婷1，胡以俊1，陈丽2，姜广水1. 慢性牙周炎和2型糖尿病患者肠道柔嫩梭菌类群的定量分析[J]. 山东大学学报(医学版), 2013, 51(4): 100-104.

LIU Wen-ting1, HU Yi-jun1, CHEN Li2, JIANG Guang-shui1. Quantitative analysis of Clostridium leptum subgroup in patients with chronic periodontitis and type 2 diabetes mellitus[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(4): 100-104.

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