INFLUENCE OF PEPTIC ULCER AND AGEING ON PROTON PUMP GENE EXPRSSION IN GASTROINTESTINAL MUCOSA

我们开发了一种简单的聚合酶链反应（PCR）和原位杂交技术，该技术灵敏度足以检测内镜下获得的胃粘膜中H^+/K^+-ATP酶（质子泵）α亚单位mRNA。β-肌动蛋白mRNA作为对照进行研究，以实现质子泵表达的半定量。应用上述方法，我们研究了奥美拉唑对6例胃溃疡患者胃体粘膜质子泵表达的影响。奥美拉唑20 mg/d治疗4周后，患者胃体粘膜标本中PCR扩增的质子泵与β-肌动蛋白的比值从1.15显著增加至2.98，原位杂交显示质子泵mRNA染色。本研究通过对消化性溃疡患者胃液中H.pylori的计数，为临床上检测少量的H.pylori mRNA和DNA提供了一种新的方法。

We developed a simple polymerase chain reaction (PCR) and in situ hybridization technique sufficiently sensitive for the detection of H^+/K^+-ATPase (proton pump) alpha subunit mRNA in endoscopically obtained gastric mucosa. The beta-actin mRNA was studied as a control to enable semiquantitation of proton pump expression. Using these methods, we studied the effect of omeprazole on proton pump expression in gastric body mucosa of 6 patients with gastric ulcer. The ratio of PCR-amplified proton pump to beta-actin was significantly increased from 1.15 to 2.98, as was staining of proton pump mRNA by in situ hybridization, in specimens of gastric body mucosa obtained from the patients after 4 weeks treatment with omeprazole 20 mg/day.Furthermore, we examined the Helicobacter pylori (H.pylori) DNA in endoscopically obtained gastric juice by competitive PCR method. We could count the number of H.pylori in gastric juice of the patients with peptic ulcer.We expected that these techniques will probe useful for the mRNAs and DNAs present in small amounts in clinical specimens.