改进的ELISA方法检测细胞角蛋白8在小鼠肠黏膜损伤模型中的作用和价值  

A modified ELISA method for cytokeratin 8 detection in the mouse small intestine mucosal injury model:role and value

作　　者：田园 尚明宇 刘俊林[2] 蔚京京[1] TIAN Yuan;SHANG Ming-yu;LIU Jun-lin;WEI Jing-jing(Department of Pediatrics,Shanxi Medical University,Taiyuan 030001,China;General Surgery Department,Liuyang City People's Hospital,Liuyang 410300,China)

机构地区：[1]山西医科大学儿科医学系,太原030001 [2]浏阳市人民医院普外科,浏阳410300

出　　处：《现代免疫学》2024年第5期396-402,共7页Current Immunology

基　　金：山西省教育厅省筹留学回国人员资助项目(2024-075);山西省大学生创新创业训练计划项目(20230278)。

摘　　要：探讨改进的ELISA检测细胞角蛋白8(cytokeratin 8,K8)在小鼠肠黏膜损伤模型中的作用和价值。本研究采用甲酸钠和H_(2)O_(2)肠内注射及小动脉夹夹闭肠系膜上动脉分支造成空回肠部分缺血等方法建立小鼠肠黏膜损伤模型,通过观察肠组织病理改变、改进的ELISA方法检测小鼠术后24 h内粪便或不同时间点血清中K8的水平,分析其变化及差异。通过绘制ROC曲线计算AUC检测该方法的真实性并分析其灵敏度和特异度筛选该检测方法的最佳截断值,为临床检测提供参考。结果发现肠内注射甲酸钠和H_(2)O_(2)和小动脉夹夹闭肠系膜上动脉分支均可造成肠黏膜损伤模型,肠组织均有明显的病理学改变;肠内注射术后24 h粪便及缺血再灌注术后不同时间点血清中K8水平升高。改进的ELISA方法真实性高,粪便和血清K8最佳截断值分别为69.78μg/mL和0.74μg/mL,灵敏度和特异度均较高。使用该方法检测小鼠粪便或血清中K8的水平可以反映肠黏膜损伤程度,为临床监测肠黏膜损伤疾病的发生判断其严重程度提供参考和依据,有较高的应用价值。An improved ELISA method to detect cytokeratin 8(K8) in the mouse intestinal mucosal injury model was evaluated in this study.The mouse intestinal mucosal injury models were established by either enteral injection of sodium formate and H_2O_(2) or via partial ischemia caused by arteriolar clamps clipping of superior mesenteric artery branches.Pathological changes in intestinal tissue were observed and an optimized ELISA method was used to detect the level of K8 in stools within 24 h after surgery or in serum at different time points,and the change and expression difference of K8 were analyzed.Next,the ROC curve and the AUC were calculated to evaluate the authenticity of the detection method.The sensitivity and specificity were also analyzed to screen for the best cut-off value to provide a reference for clinical detection.The results showed that both enteral injection of sodium formate and H_2O_(2) and arteriolar clamps clipping superior mesenteric artery branches caused significant pathological changes in intestinal tissue.The levels of K8 increased in stools within 24 h after surgery or in serum at different time points after ischemia and reperfusion.The improved ELISA method has high authenticity,and the optimal cut-off values for fecal and serum K8 are 69.78 μg/mL and 0.74 μg/mL respectively,with both high sensitivity and specificity.Utilizing this method to detect the level of K8 in feces or serum of mice reflected the degree of intestinal mucosal damage,therefore providing a reference for clinical monitoring of the occurrence and severity of intestinal mucosal injury,and has high application value.

关 键 词：酶联免疫吸附法 肠黏膜损伤 细胞角蛋白 

分 类 号：R446.6[医药卫生—诊断学]