Department of Internal Medicine, Inha University School of Medicine, Incheon, Korea.
Gut Liver. 2010 Mar;4(1):60-7. doi: 10.5009/gnl.2010.4.1.60. Epub 2010 Mar 25.
BACKGROUND/AIMS: Bacterial infection is accepted as a precipitating factor in cholesterol gallstone formation, and recent studies have revealed the presence of Helicobacter species in the hepatobiliary system. We utilized the polymerase chain reaction (PCR) to establish the presence of bacterial DNA, including from Helicobacter species, in gallstones, bile juice, and gallbladder mucosa from patients with gallstones.
At cholecystectomy, 58 gallstones, 48 bile samples, and 46 gallbladder mucosa specimens were obtained and subjected to nested PCR using specific 16S rRNA primers of H. pylori and other bacteria. Bacterial species were identified by DNA sequencing analysis. Bacterial 16S rRNA was detected in 25 out of 36 mixed-cholesterol gallstones, 1 out of 10 pure-cholesterol gallstones, and 9 out of 12 pigmented stones. Furthermore, 16S rDNA sequencing identified Escherichia coli, Pseudomonas, Citrobacter, Klebsiella, and Helicobacter species.
Helicobacter DNA was detected in 4 out of 58 gallstones, 6 out of 48 bile samples, and 5 out of 46 gallbladder specimens. Direct sequencing of Helicobacter amplicons confirmed strains of H. pylori in all four gallstones, five out of six bile samples, and three out of five gallbladder specimens. Almost all mixed-cholesterol gallstones appear to harbor bacterial DNA, predominantly E. coli.
H. pylori was also found in the biliary system, suggesting that these bacteria are of etiological importance in gallstone formation.
背景/目的:细菌感染被认为是胆固醇结石形成的诱发因素,最近的研究表明,在肝胆系统中存在幽门螺杆菌属。我们利用聚合酶链反应(PCR)来确定胆结石、胆汁和胆结石患者的胆囊黏膜中是否存在细菌 DNA,包括幽门螺杆菌属。
在胆囊切除术中,获得了 58 个胆结石、48 个胆汁样本和 46 个胆囊黏膜标本,并使用幽门螺杆菌和其他细菌的特异性 16S rRNA 引物进行嵌套 PCR。通过 DNA 测序分析鉴定细菌种类。在 25 个混合胆固醇胆结石、1 个纯胆固醇胆结石和 12 个色素结石中检测到细菌 16S rRNA。此外,16S rDNA 测序鉴定出大肠埃希菌、假单胞菌、柠檬酸杆菌、克雷伯菌和幽门螺杆菌属。
在 58 个胆结石中检测到 4 个、48 个胆汁样本中检测到 6 个、46 个胆囊标本中检测到 5 个幽门螺杆菌属 DNA。幽门螺杆菌属扩增子的直接测序在 4 个胆结石、6 个胆汁样本和 5 个胆囊标本中均证实了 H. pylori 株的存在。几乎所有的混合胆固醇胆结石都似乎携带细菌 DNA,主要是大肠埃希菌。
在胆道系统中也发现了 H. pylori,这表明这些细菌在胆结石形成中具有重要的病因学意义。