光学显微镜食管下段细胞间隙的测量对胃食管反流病诊断的意义

A study on the light microscopic measurement of intercellular space of squamous epithelium in lower- esophagus to diagnose gastroesophageal reflux disease

目的通过光学显微镜（光镜）对胃食管反流病（GERD）患者食管下段细胞间隙的测量，探讨光镜下细胞间隙的变化对GERD的诊断意义。方法158例受试者行内镜检查，分为正常对照组（25例）、非糜烂性反流病（NERD）组（58例）和糜烂性食管炎（EE）组（75例），在齿状线以上2～3cm取活检行光镜检查，并随机选取43例（其中正常对照组4例，EE组28例，NERD组11例）在相同部位取活检同时进行电镜检查。光镜／电镜下观察切片并采集图像，采用图像分析系统对每例切片测量100个细胞间隙，并进行统计学分析。结果正常对照组、NERD组及EE组光镜下测量的平均细胞间隙（LMIS）分别为（0．61±0．10）μm、（1．12±0．61）μm和（1．30±0．19）μm，3组两两相比差异均有统计学意义（P〈0．05）；EE组不同洛杉矶分级的各亚组间相比，细胞间隙增宽程度的差异无统计学意义（P〉0．05）；NERD患者食管24hpH监测阳性者（40例）与阴性者（17例）LMIS的差异无统计学意义（P〉0．05）；光镜下LMIS的截断（cut—off）值为0．85μm，如果以临床症状加内镜和食管24hpH监测综合作为诊断GERD的金标准，LMIS诊断GERD的敏感性为95．5％，特异性为100．0％；同时做光镜和电镜测量的受试者共43例，经相关性分析和一致性检验，光镜测量和电镜测量的细胞间隙宽度显著相关（r=0．737，P=0．000），且一致性好。结论GERD患者食管下段组织的细胞间隙可通过光镜进行定量测量，其结果与电镜测量结果的一致性好。光镜下细胞间隙增宽可以作为GERD诊断的敏感、特异且较为客观的指标之一。

Objective To evaluate the significance of dilated intercellular space （DIS） of squamous epithelium in lower-esophagus which was measured at light microscopy （LM） in the diagnosis of gastroesophageal reflux disease （GERD）. Methods This study was divided into two parts. The first part consists of 133 GERD patients with typical symptoms, including 75 erosive esophagitis （EE） and 58 non- erosive reflux disease （NERD） ; the second part consists of 25 healthy volunteers as control; ambulatory 24-hr esophageal pH monitoring, endoscopy and biopsies which were taken in lower esophagus were performed in all individuals. NERD patients with negative ambulatory 24-hr esophageal pH monitoring were undertaken PPI test. We selected 43 individuals randomly （4 control, 11 NERD and 28 EE） to measure the intercellular space diameter with LM and transmission electronic microscopy （TEM） simultaneously. All the samples were observed at immersion objective, taken pictures and make the scale at the same time; all the pictures were measured by the computer-assisted morphometry and in the samples for one case should be measured 100 points and then calculate the mean intercellular space diameter, while the TEM has the same procedures. Results A total of 158 individuals （90 male, 68 female） were enrolled in this study. The mean intercellular space diameter with LM of the controls, the NERD and the EE patients were （0. 61 ± 0. 10） μm, （ 1.12 ±0. 61 ） μm and （ 1.30 ±0. 19） μm, respectively, with significant differences between the control and NERD group, the control and EE group, the EE and NERD group （ P 〈 0. 05 ） ; the mean intercellular space diameter with LM of EE subgroup of LA-A, LA-B, LA-C, LA-D were （ 1.31 ± 0. 23） μm, （ 1.27 ±0. 17） μm, （ 1.31±0. 14） μm, （1.33 ±0. 11）μm, respectively, without significant differences among every two subgroups （ P 〉 0. 05 ） ; NERD patients with positive and negative 24-hr ambulatory pH monitoring were 40 and 17 cases, and the mean intercellular space diameter with LM were （ 1.12 ±0. 16 ） μm, （ 1.11 ± 0. 31 ） μm, respectively, without significant differences between them （ P 〉 0. 05） ; the cut-off value of mean intercellular space diameter with LM was 0. 85 μm, if combination the clinical GERD symptom, endoscopy and 24-hr ambulatory pH monitoring as gold standard, its sensitivity was 95.5% and specificity was 100% ; the intercellular space diameter of the controls, the NERD patients and the EE patients with TEM were （0. 30 ±0. 14） μm, （ 1.33 ±0. 28） μm, （ 1.40 ±0. 22） μm, respectively, with significant differences between the control and NERD group, the control and EE group （P 〈 0. 05）. It was significant correlation in intercellular space diameter between LM and TEM （ r = 0. 737, P = 0. 000 ）. Conclusions The intercellular space diameter of squamous epithelium in lower esophagus of GERD patients can make quantitative study with LM. It is significant correlation with TEM. DIS with LM is one of sensitive, specific and objective marker of GERD.