Department of Microbiology and Clinical Microbiology, Gastroenterology Clinic, Medical Faculty, Atatürk University, Erzurum, Turkey.
J Clin Gastroenterol. 2010 Jul;44(6):416-22. doi: 10.1097/MCG.0b013e3181c21c69.
BACKGROUND-GOALS: We used transmission electron microscopy and immunohistochemistry (IHC) to investigate how Helicobacter pylori affects the gastric mucosa of humans.
Gastric biopsy specimens were obtained from 15 patients with gastric discomfort. The samples were processed using both microscopic examinations and a real-time polymerase chain reaction to detect H. pylori DNA. IHC staining was performed with an avidin-biotin complex immunoperoxidase kit for paraffin-embedded tissue sections. Polyclonal rabbit anti-H. pylori was used as a primary antibody.
IHC-applied slides with brown-stained spiral bacteria on the luminal surface and in the intercellular spaces of the gastric epithelium; electron-dense spiral H. pylori of approximately 200 to 300 nm in diameter both in the gastric lumen and between the gastric epithelial cells; coccoid or ellipsoid H. pylori attached to the epithelial cells through egg-cup-like pedestals; coccoid H. pylori within the endocytotic vesicles in the apical cytoplasmic part of the epithelial cells, thus suggesting their internalization by phagocytosis; electron-dense spiral H. pylori within the membrane-bounded vacuoles of both the gastric epithelial cells, and the lamina propria; a prominent vacuolization of gastric epithelial cells invaded by H. pylori; and swollen and lytic gastric epithelial cells that suggest a mucosal erosion and may lead to peptic ulcer. All of these microscopic findings were not present in the H. pylori DNA-negative specimens that were used as the control group.
This is the first histomicrobiologic study to show gastric cells invaded by H. pylori in patients with H. pylori infection confirmed by real-time polymerase chain reaction.
本研究采用透射电子显微镜和免疫组织化学(IHC)方法,观察幽门螺杆菌(H. pylori)感染对人类胃黏膜的影响。
收集 15 例有胃部不适症状的患者的胃活检标本。通过显微镜检查和实时聚合酶链反应(PCR)检测 H. pylori DNA 对标本进行处理。采用链霉菌抗生物素蛋白过氧化物酶法对石蜡包埋组织切片进行 IHC 染色,使用多克隆兔抗 H. pylori 作为一抗。
IHC 应用载玻片上可见在胃上皮细胞的腔面和细胞间隙中存在棕褐色染色的螺旋状细菌;在胃腔和胃上皮细胞之间均可见直径约 200-300nm 的电子致密螺旋状 H. pylori;通过杯状基底部附着在上皮细胞的球形或椭圆形 H. pylori;在上皮细胞顶质部分的内吞小泡中存在球形 H. pylori,提示其通过吞噬作用内化;在胃上皮细胞和固有层的膜结合空泡中存在电子致密的螺旋状 H. pylori;被 H. pylori 侵袭的胃上皮细胞出现明显的空泡化;肿胀和裂解的胃上皮细胞提示黏膜糜烂,可能导致消化性溃疡。这些微观发现均不存在于作为对照组的 H. pylori DNA 阴性标本中。
这是第一项通过实时聚合酶链反应证实 H. pylori 感染的患者胃细胞中存在被 H. pylori 侵袭的组织微生物学研究。