改良构建大鼠单侧输尿管梗阻肾间质纤维化模型

Improvement for the construction of renal interstitial fibrosis model with unilateral ureteral obstruction in rats

背景:传统的大鼠单侧输尿管梗阻手术方法,可以在较短的时间内(一两周)导致动物梗阻侧肾脏发生肾间质纤维化,但具有术后并发症较多、死亡率较高的缺点。目的:优化并改良大鼠单侧输尿管梗阻手术操作,降低动物术后的并发症发生率,提高存活率,检测改良单侧输尿管梗阻方法建模大鼠的病理生理指标,为功能药理学基础研究提供背景数据支持。方法:①取20只雄性SD大鼠,随机分为4组,分别进行传统单侧输尿管梗阻方法结扎、改良单侧输尿管梗阻方法结扎、不游离输尿管结扎、开腹腔但不结扎输尿管(假手术组)操作,术后14 d时比较组间大鼠肾间质纤维化成模率、死亡率、术后并发症发生率的差异;②另取28只SD大鼠,雌雄各半,随机分为改良组与假手术组,采用尿液分析、血细胞分析、血液生化分析方法,检测两组大鼠的生理指标,并通过苏木精-伊红染色与Masson染色法,观察改良组大鼠术后14,21 d时的肾脏组织病理学结果。结果与结论:①与不游离输尿管结扎组大鼠相比,传统单侧输尿管梗阻组和改良单侧输尿管梗阻组大鼠的成模率高;②在单侧输尿管梗阻手术操作过程中,采用输尿管中段双结扎、中间不剪断的改良方法与传统方法相比,操作更简便易行,手术开创更小,肾间质纤维化成模快、成模率较高,动物术后死亡率与并发症发生率均低;③改良组与假手术组相比,术后14 d时的尿素氮、谷草转氨酶、白蛋白等血液生化指标差异有显著性意义(P<0.05),其梗阻侧肾脏的组织病理切片苏木精-伊红染色与Masson染色结果均可见典型的肾间质纤维化病理特征。

BACKGROUND:Unilateral ureteral obstruction(UUO)is a traditional surgical method that can cause renal interstitial fibrosis in rats in a short period of time(1-2 weeks),but it can develop many postoperative complications and result in a high mortality.OBJECTIVE:To optimize and improve the UUO operation in rats for reducing the incidence of postoperative complications and improving the survival rate of animals,and to detect the pathophysiological indicators of rats modeled by UUO,providing background data for the basic research on functional pharmacology.METHODS:Twenty male Sprague-Dawley rats were randomly divided into four groups,followed by traditional UUO ligation,improved UUO ligation,undissociated ureteral ligation,and opening the abdominal cavity with no ureter ligation(sham group),respectively.At 14 days postoperatively,the successful rate of renal interstitial fibrosis model,mortality rate,and the incidence of postoperative complications were compared between groups.Another 28 Sprague-Dawley rats,half male and half female,were randomly divided into improved UUO group and control group.Physiological indexes,including urine analysis,blood cells analysis,blood biochemical analysis,were tested as background information.Besides,the pathological changes of kidney tissues were compared between groups using hematoxylin-eosin staining and Masson staining at 14 and 21 days postoperatively.RESULTS AND CONCLUSION:Compared with the undissociated ureteral ligation group,the successful rate of modeling was higher in the traditional and improved UUO groups.The improved UUO surgery with double ligation of the middle ureter and no cutting of the middle ureter was characterized by easier operation,smaller surgical wound,faster modeling and higher successful rate of renal interstitial fibrosis model,lower postoperative mortality and lower incidence of complications in animals as compared with the traditional UUO surgery.The improved UUO group showed statistically significant differences from the sham group in blood biochemical indexes such as urea nitrogen,glutamate aminotransferase,and albumin(P<0.05).Results from hematoxylin-eosin staining and Masson staining of the obstructed kidney revealed typical pathological features of renal interstitial fibrosis.