共聚焦激光内镜观察急性胰腺炎微循环的实验研究

Observation of acute pancreatitis microcirculation with confocal laser endoscopy

目的探讨共聚焦激光内镜（CLE）在活体观察急性胰腺炎（AP）微循环变化的价值。方法将32只SD大鼠随机分为：对照组8只，进腹后仅做相应的翻动，找出胃、十二指肠、胰腺后，充分暴露胰腺，找到下腔静脉采血后按照4ml／kg注射0．5％荧光素钠，注射完毕时以CLE观察胰腺组织并记录；实验组24只，进腹结扎胆总管末端制作AP模型，分别在AP模型制作完成后12h（12h组，8只）、24h（24h组，8只）、48h（48h组，8只）再次开腹观察胰腺，找到下腔静脉采血后注射荧光素钠，注射完毕时以CLE观察病变胰腺组织并记录。各组均取胰腺组织做病理切片。结果运用CLE可以观察到正常胰腺腺泡结构和毛细血管以及血管中流动的红细胞，并有可能数出一时段内通过某一毛细血管横切面的红细胞个数；12h组腺泡结构基本正常，微循环毛细血管内可见红细胞流动但流速明显减慢；24h组部分腺泡结构显影模糊，但仍可见结构基本正常的腺泡，能够观察到的微循环毛细血管明显减少，毛细血管内红细胞数量明显减少且流速很慢；48h组腺泡显影模糊，部分腺泡不显影，几乎观察不到正常腺泡结构，极少能够观察到微循环毛细血管，且血管内罕见红细胞，几乎无红细胞流动。CLE未能观察到确切的胰岛组织，难以辨识白细胞，不能判断是否有微血栓形成。结论CLE对胰腺微循环研究有较大价值，特别是对活体AP微循环血流状态的判断较以往实验方法具明显优势，但仍需要改进显色剂和图像处理技术以获得更易辨识的图像，提升其应用价值。

Objective To study microcirculation of acute pancreatitis （AP） in vivo with confocal laser endoscopy （CLE）. Methods Thirty-two SD rats were randomly divided into 2 groups,i, e. control（n=8） and experiment group（ n=24）. In the control group, pancreas were identified, the blood was collected from the inferior vena cava, 0. 5% uranin was administered at 4 ml/kg intravenously, and the pancreas were observed through CLE. In the experiment group, AP model was established by ligation of the end of common bile duct. The abdomen was opened to observe the pancreas at 12 h （ n = 8 ） ,24 h （ n = 8 ） and 48 h （ n = 8 ） , uranin was administered intravenously, and the pancreatic tissue was observed through CLE. Pancreatic tissue in each group was sent for pathological study. Results Both healthy and affected pancreatic microcircu-lation can be viewed through CLE. The number of red blood ceils can be counted within certain period of time cross-sectionally. In 12 h group,the construction of pancreatic acini was normal, and the flow of red blood ceils could be seen in the capillary, but quite slow. In 24 h group, the structure was vague, but there were still normal pancreatic acini less visible capillaries and less and slower red blood ceils flow were observed. In 48 h group, some pancreatic acini were not visible, normal pancreatic acinar structure and microcirculation could not be observed, neither could red blood ceils. Insulin tissue, white blood ceils and microthrombosis could not be observed through CLE. Conclusion CLE is distinctly valuable for pancreatic microcirculation study, especially for AP microcirculation in vivo. However, color developing agents and image processing techniques should be improved io ensure belier images and better application.