State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, People's Republic of China.
Microb Ecol. 2012 Feb;63(2):304-13. doi: 10.1007/s00248-011-9925-5. Epub 2011 Aug 4.
The beneficial effects of Bifidobacteria on health have been widely accepted. Patients with chronic liver disease have varying degrees of intestinal microflora imbalance with a decrease of total Bifidobacterial counts. Since different properties have been attributed to different Bifidobacterium species and there is no information available for the detailed changes in the genus Bifidobacterium in patients with chronic liver disease heretofore, it is meaningful to investigate the structure of this bacterium at the species level in these patients. The aim of this study was to characterize the composition of intestinal Bifidobacterium in patients with hepatitis B virus-induced chronic liver disease. Nested-PCR-based denaturing gradient gel electrophoresis (PCR-DGGE), clone library, and real-time quantitative PCR were performed on the fecal samples of 16 patients with chronic hepatitis B (CHB patients), 16 patients with hepatitis B virus-related cirrhosis (HBV cirrhotics), and 15 healthy subjects (Controls). Though there was no significant difference in the diversity among the three groups (P = 0.196), Bifidobacterium dentium seems to be specifically enhanced in patients as the PCR-DGGE profiles showed, which was further validated by clone library and real-time quantitative PCR. In contrast to the B. dentium, Bifidobacterium catenulatum/Bifidobacterium pseudocatenulatum were detected less frequently in the predominant profile and by quantitative PCR in HBV cirrhotics than in the controls, and the level of this species was also significantly different between these two groups (P = 0.023). Although having no quantitative difference among the three groups, Bifidobacterium longum was less commonly detected in HBV cirrhotics than in CHB patients and Controls by quantitative PCR (P = 0.011). Thus, the composition of intestinal Bifidobacterium was deeply altered in CHB and HBV cirrhotic patients with a shift from beneficial species to opportunistic pathogens. The results provide further insights into the dysbiosis of the intestinal microbiota in patients with hepatitis B virus-induced chronic liver disease and might potentially serve as guidance for the probiotics interventions of these diseases.
双歧杆菌对健康的有益影响已被广泛接受。慢性肝病患者的肠道微生物群存在不同程度的失衡,双歧杆菌总数减少。由于不同的双歧杆菌种具有不同的特性,并且迄今为止尚无关于慢性肝病患者双歧杆菌属详细变化的信息,因此在这些患者中研究该细菌的种水平结构具有重要意义。本研究旨在描述乙型肝炎病毒引起的慢性肝病患者肠道双歧杆菌的组成。对 16 例慢性乙型肝炎(CHB 患者)、16 例乙型肝炎病毒相关肝硬化(HBV 肝硬化患者)和 15 例健康对照者的粪便样本进行基于嵌套 PCR 的变性梯度凝胶电泳(PCR-DGGE)、克隆文库和实时定量 PCR。尽管三组之间的多样性没有显著差异(P=0.196),但 PCR-DGGE 图谱显示双歧杆菌 dentium 似乎在患者中特异性增强,这进一步通过克隆文库和实时定量 PCR 得到验证。与 B. dentium 相反,在主要图谱和定量 PCR 中,HBV 肝硬化患者中双歧杆菌 catenulatum/Bifidobacterium pseudocatenulatum 的检出频率低于对照组,并且这两个组之间的该种水平也存在显著差异(P=0.023)。尽管三组之间没有定量差异,但在 HBV 肝硬化患者中,Bifidobacterium longum 的检出频率低于 CHB 患者和对照组,定量 PCR 也存在显著差异(P=0.011)。因此,CHB 和 HBV 肝硬化患者的肠道双歧杆菌组成发生了深刻改变,从有益种转变为机会性病原体。该结果进一步深入了解了乙型肝炎病毒引起的慢性肝病患者肠道微生物群的失调,并可能为这些疾病的益生菌干预提供指导。