急性胰腺炎大鼠外周血淋巴细胞亚群的变化

摘要

目的检测急性胰腺炎（ AP）大鼠外周血淋巴细胞亚群，探讨AP病程进展过程中免疫状态改变的原因。方法60只SPF级雄性Wistar大鼠随机分为空白组、假手术组、AP组（ AP 6 h组、12 h组及AP 24 h组）。采用胰腺被膜下注射牛磺胆酸钠（ Na-Tc）建立急性胰腺炎模型。空白组及假手术组在第6小时处死，胰腺炎组在造模后第6、12、24小时3个时点分批处死。每只大鼠处死前腹主动脉抽血5 ml，取胰腺组织行HE染色并做病理评分。测定每只大鼠淋巴细胞计数及血清淀粉酶，用流式细胞仪测定淋巴细胞亚群CD3＋、CD4＋、CD8＋、CD19＋及CD56＋。结果假手术组与空白组比较淋巴细胞无明显降低（ P ＞0构．05），随着病程的延长CD3＋淋巴细胞逐渐下降，CD8＋淋巴细胞轻度增高，CD4＋/CD8＋明显下降，CD3＋/CD19＋明显上升（ P ＜0．05）。结论在AP病程中免疫系统的紊乱是逐渐加重的。%Objective To investigate the changes of immune functional status in the development of acute pancreatitis ( AP) by examining lymphocyte subsets of peripheral blood in rats with acute pancreatitis .Methods Sixty specific pathogen free ( SPF) male Wistar rats were randomly divided into five groups:blank group ( n =11),sham-operation group ( n =11),acute pancreatitis 6-hour group (AP 6h group, n =11),acute pancreatitis 12-hour group (AP 12h group, n =13),acute pancreatitis 24-hour group (AP 24h group, n =14).The animal models with acute pancreatitis wee established by injecting 5% Na-Tc into capsula pancreatic .The rats in blank group and sham-operation group were killed at 6 hour after the operation , however , the rats in the three acute pancreatitis groups were sacrificed respectively at 6,12 and 24 hours after the models were established .The content of amylase in serum and the counts of lymphocytes were detected by automatic biochemistry analyzer .The changes in the rats'pancreatic tissues were observed by Hematoxylin-Eosin ( HE) staining and were pathologically scored . Lymphocyte subsets (CD3,CD4,CD8,CD19,CD56) in peripheral blood were measured by flow cytometry .Results There were no significant differences in lymphocyte counts between sham-operation group and blank group ( P >0.05),however,with the development of course of disease the CD 3 +lymphocytes were gradually decreased ,but CD8 +lymphocytes were slightly increased , and the ratio of CD4 +/CD8 +was obviously decreased , however , the ratio of CD3 +/CD19 + was significantly increased .Conclusion The disorder of immune system is gradually aggravated with the development of AP .