大鼠肝移植术后脓毒症模型的构建和分析

Construction and analysis of a sepsis model of rat after liver transplantation

  • 摘要:
    目的  建立一种用于临床转化研究的稳定可靠的大鼠肝移植(LT)术后脓毒症模型并分析其特征。
    方法  采用“二袖套法”建立SD大鼠原位LT模型,于术后3 d通过盲肠结扎穿孔(CLP)构建脓毒症模型。将SD大鼠随机分为3组:假手术组(Sham组)、LT组、LT + CLP组,每组各6只。比较各组体质量、肛温、生存率的变化,通过脓毒症评分进行评估。检测各组血液生化指标丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、尿素(Urea)、肌酐(Cr)、肌酸激酶(CK)、乳酸脱氢酶(LDH)与炎症因子白细胞介素(IL)-1β、IL-6、IL-10、肿瘤坏死因子(TNF)-α水平,以及观察不同器官组织病理学与细胞凋亡变化。
    结果  与Sham组相比,LT组和LT+CLP组体质量均下降(均为P<0.05)。LT+CLP组肛温在术后呈不断下降趋势、脓毒症评分在术后急剧升高、生存率降至16.7%,与Sham组和LT组间的差异均有统计学意义(均为P<0.05)。LT + CLP组大鼠术后72 h内ALT、AST、Urea、Cr、CK、LDH及血清IL-1β、IL-6、IL-10、TNF-α水平均高于Sham组和LT组大鼠(均为P<0.05)。LT + CLP组病理学检查显示多器官存在严重的组织结构破坏、坏死及炎症细胞浸润,脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记(TUNEL)染色显示多器官细胞凋亡水平升高。
    结论  利用大鼠肝移植联合CLP成功构建了稳定的肝移植感染动物模型,表现出高病死率、显著多器官损伤和剧烈的炎症反应,为移植感染研究提供了理想的动物模型。

     

    Abstract:
    Objective  To establish a stable and reliable sepsis model of rat after liver transplantation (LT) for clinical translational research and analyze its characteristics.
    Methods  The "two-sleeve method" was used to establish the in situ LT model of SD rats, and the sepsis model was constructed through cecal ligation and puncture (CLP) at 3 d after the operation. SD rats were randomly divided into 3 groups: sham operation group (Sham group), LT group, and LT + CLP group, with 6 rats in each group. The changes in body weight, rectal temperature and survival rate were compared, and the sepsis score was used for evaluation. The levels of blood biochemical indicators alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea (Urea), creatinine (Cr), creatine kinase (CK), lactate dehydrogenase (LDH) and inflammatory factors interleukin (IL)-1β, IL-6, IL-10, tumor necrosis factor (TNF)-α in each group were detected, and the pathological changes and cell apoptosis in different organs were observed.
    Results  Compared with the Sham group, the body weight of the LT group and LT + CLP group decreased (all P<0.05). The rectal temperature of the LT + CLP group showed a continuous downward trend after the operation, the sepsis score increased sharply after the operation, and the survival rate dropped to 16.7%, and the differences between the Sham group, LT group and LT + CLP group were statistically significant (all P<0.05). The levels of ALT, AST, Urea, Cr, CK, LDH, and serum IL-1β, IL-6, IL-10 and TNF-α in the LT + CLP group were higher than those in the Sham group and LT group rats within 72 hours after the operation(all P<0.05). The pathological examination of the LT + CLP group showed severe tissue structure destruction, necrosis and infiltration of inflammatory cells in multiple organs, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining showed an increased level of cell apoptosis in multiple organs.
    Conclusions Using liver transplantation combined with CLP, a stable animal model of liver transplantation infection is successfully established, which exhibits a high mortality rate, significant multi-organ damage and intense inflammatory response, providing an ideal animal model for transplantation infection research.

     

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