关键词: 酒精性损伤, 探头式激光共聚焦显微镜, 消化道, 实时观察, 高分辨率, 小鼠

Abstract:

Objective To explore the feasibility of applying probe-based confocal laser endomicroscopy (pCLE) in rapidly detecting and evaluating the morphological characteristics of digestive tract tissues in animal experiments. Methods 6 male Kunming mice of SPF were modeled by gastric gavage with 52 ° Red Star Erguotou, and 6 were performed by gastric gavage with drinking water at room temperature. After 28 days of modeling, 3 mice were randomly selected from each group. After being anesthetized by intraperitoneal injection of 10% urethane, the stomach, duodenum, jejunum and cecum tissues were excised and immersed in 1% fluorescein sodium dye for staining.The microstructure of the mucosal surface of the gastrointestinal tissue was observed by pCLE. The rest of the mice in the model group and the control group were performed by cardiac perfusion, and the gastric, duodenum, jejunum, and rectum tissue were stained with Hematoxylin-Eosin (HE). The microstructure of gastric and intestinal tissue was observed under microscope. Results Under the field of pCLE,the fluorescence staining of the tissue on the surface of the digestive tract mucosa was uniform and the shapes of the gastric pits, intestinal papillae, and intestinal crypts were complete, closely arranged, and the edge structures were clear in the control group. In the model group, the mucosal tissues of the digestive tract showed swelling and deformation, uneven fluorescence staining, fluorescein leakage, defects in some tissues or cell shedding, and unclear boundaries between the characteristic structures of adjacent tissues. Some tissues had defects or cell shedding, and the boundaries between the characteristic structures of adjacent tissues were unclear. HE staining showed that the digestive tract tissue structure in the control group was normal, neatly arranged, without defects. The sizes of the submucosal glands were consistent without hyperplasia, and there was no obvious inflammatory cell infiltration. In the model group, some of the digestive tract tissue structures were defective, sparsely arranged, the submucosal glands were atrophied, accompanied by inflammatory cell infiltration. The histological characteristics observed by in vivo pCLE were consistent with those of ex vivo HE staining. Conclusion pCLE can realize rapid, real-time, large-scale and high-resolution microscopic imaging of gastrointestinal mucosa, and more realistically and comprehensively display the microstructural characteristics under physiological and pathological conditions. It demonstrates substantial application potential and promising prospects in the experimental investigations of histological assessment for digestive system injuries.

Key words: Alcoholic injury, Probe-based Confocal Laser Endomicroscopy, Digestive tract, Real-time observation, High resolution, Mice