Kobayashi Takashi, Harada Kenichi, Miwa Koichi, Nakanuma Yasuni
Department of Human Pathology, Kanazawa University Graduate School of Medicine, Kanazawa, Japan.
Dig Dis Sci. 2005 May;50(5):862-7. doi: 10.1007/s10620-005-2654-1.
Several Helicobacter species are known to colonize the biliary tract in animals and have been identified in the gallbladder bile of a high proportion of Chilean patients with gallbladder cancer. In this study, we tried to examine the presence of Helicobacter species in the bile to know their participation in the development of extrahepatic biliary diseases. DNA was extracted from 57 bile samples from 30 patients with benign biliary diseases (cholecystolithiasis and choledochocystolithiasis), 6 malignant biliary diseases (gallbladder cancer and common bile duct cancer), and 21 nonbiliary diseases. The presence of Helicobacter genus-, H. pylori-, H. hepaticus-, and H. bilis-specific 16S rRNA genes, the H. pylori urease A gene, and the H. pylori 26K protein gene in the bile was determined by PCR and sequencing analysis. Helicobacter genus DNA (shorter amplicons, 400 bp) was statistically frequently detected in biles from 53% (16/30) and 86% (5/6) of benign and malignant biliary diseases, compared with 9% (2/21) of nonbiliary diseases, but longer amplicons (1200 bp) were not detectable in any samples. The H. pylori urease A gene (nested amplicon) was also frequently found in bile, whether benign, malignant, or control, though neither H. pylori 16S rRNA nor the 26K protein gene was detectable in any bile samples. H. bilis-16S rRNA genes were detectable in only two cases. H. hepaticus was not detectable in any samples. DNA fragments of Helicobacter species other than H. pylori, H. hepaticus, and H. bilis are commonly detectable in the bile of patients with extrahepatic biliary diseases, whether benign or malignant, implying that the Helicobacter genus may be directly or indirectly involved in the pathogenesis of these diseases.
已知几种幽门螺杆菌可在动物的胆道中定殖，并且在很大比例的智利胆囊癌患者的胆囊胆汁中已被鉴定出来。在本研究中，我们试图检测胆汁中幽门螺杆菌的存在情况，以了解它们在肝外胆道疾病发生发展中的作用。从30例良性胆道疾病（胆囊结石和胆总管结石）、6例恶性胆道疾病（胆囊癌和胆总管癌）患者以及21例非胆道疾病患者的57份胆汁样本中提取DNA。通过PCR和测序分析确定胆汁中幽门螺杆菌属、幽门螺杆菌、肝螺杆菌和胆螺杆菌特异性16S rRNA基因、幽门螺杆菌脲酶A基因以及幽门螺杆菌26K蛋白基因的存在情况。与9%（2/21）的非胆道疾病患者相比，幽门螺杆菌属DNA（较短扩增子，400 bp）在53%（16/30）的良性胆道疾病和86%（5/6）的恶性胆道疾病患者的胆汁中被统计学频繁检测到，但在任何样本中均未检测到较长的扩增子（1200 bp）。幽门螺杆菌脲酶A基因（巢式扩增子）在胆汁中也经常被发现，无论胆汁是良性、恶性还是对照样本，尽管在任何胆汁样本中均未检测到幽门螺杆菌16S rRNA和26K蛋白基因。仅在两例样本中检测到胆螺杆菌16S rRNA基因。在任何样本中均未检测到肝螺杆菌。除幽门螺杆菌、肝螺杆菌和胆螺杆菌外，其他幽门螺杆菌种的DNA片段在肝外胆道疾病患者的胆汁中普遍可检测到，无论疾病是良性还是恶性，这意味着幽门螺杆菌属可能直接或间接参与这些疾病的发病机制。
