心导管留置术后盲肠结扎穿孔术(CLP)脓毒症动物模型的建立

Combination of Heart Catheterization with Cecal Ligation and Puncture in Setting up a Septic Model in Animals

目的采用留置心导管采血法检测盲肠穿孔术(CLP)后脓毒症模型大鼠血清学指标并观察肝肺组织病理学形态的变化,为用于治疗脓毒症药物的活性评价的动物模型的建立奠定了实验基础。方法采用改良的心导管留置术结合盲肠结扎穿孔术(CLP)建立脓毒症大鼠模型,术前和术后2、6、12、24、48 h共6个时相点采血,动态浊度法鲎试验检测血浆内毒素(LPS)水平,ELISA法检测血浆TNF-α和IL-6值,全自动血细胞分析仪检测全血白细胞(WBC)及血小板(PLT)值,术后48 h观察大鼠肝、肠、肺的病理学形态改变。结果术后大鼠的血浆LPS、TNF-α、IL-6及全血WBC水平均于术后6 h后逐渐升高,12 h达到高峰,之后逐渐下降,于48 h达到正常水平;PLT水平6 h内无明显变化,随后逐渐下降,24 h达最低值,之后逐渐升高,于48 h接近正常水平;肝、肠、肺组织则呈现以肿胀、炎性细胞浸润为主的病理学改变。结论通过心导管留置术采血观察CLP模型大鼠血细胞和细胞因子、LPS的动态变化是一稳定可靠的实验方法。

Objective To detect changes of plasma levels of serological indexes and observe pathologic differences in tissues of rats before and after cecal ligation and puncture（CLP） so as to provide refference statistics for bioactivity assessment of therapeutic agents for sepsis.Method The rat model was established by combining heart catheterization with CLP.Blood samples were obtained before and at 2 h,6 h,12 h,24 h and 48 h after CLP.Plasma levels of lipopolysacchride（LPS） were detected by the limulus test（dynamic nephelometry）.The plasma TNF-α and IL-6 were measured by enzyme-linked immunosorbent assay（ELISA） and blood WBC and PLT counts were examined by automatic blood analyzer.Pathological changes of organs such as liver,intestine and lung were also observed at 48 h after CLP.Result Levels of plasma markers LPS TNF-α and IL-6 and WBC counts increased at 6 h after CLP,reached peak at 12 h,then decreased and returned to normal at 48 h.PLT counts showed little differences within 6 h after CLP,then declined,reached the lowest level at 24 h and restored to normal at 48 h.Tissue edema and inflammatory cell infiltration were typical pathological changes in the tissues of liver,intestine and lung in CLP rats.Conclusion Combination of heart catheterization with CLP provides a novel way to improve animal models of sepsis in which stable and reliable observations are ensured.