Yang Jing, Huang Jian, Zhang Yao-zong, Chen Lin
State Key Laboratory of Trauma, Burns and Combined Injury, Trauma Center, Trauma Laboratory, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042, China.
Chin J Traumatol. 2010 Oct 1;13(5):308-12.
To evaluate the endothelial cell damage by detecting the circulating Tie2 mRNA level in a rat model of sepsis.
The model of sepsis was established by cecal ligation and puncture (CLP) in 90 rats which were divided into 6 groups: normal, sham, CLP-3 h, CLP-6 h, CLP-12 h and CLP-24 h. Serum biochemical markers were detected by automatic biochemical analyzer. Serum IL-6 was measured with enzyme linked immunosorbent assay. The vascular permeability of liver, kidney, lung and heart was detected with Evans blue. Circulating endothelial cells (CEC) were separated using density gradient separation and counted. Total RNA of whole blood were extracted and the mRNA levels of two endothelial specific genes, Tie2 and vascular endothelial growth factor receptor 2 (VEGFR2), were measured by quantitative real-time PCR.
The level of serum biochemical indexes increased after CLP. The amount of serum IL-6 in CLP-6 h, 12 h, and 24 h group was increased 6.5-fold (P <0.05), 8.4-fold (P < 0.01), and 13.3-fold (P < 0.001) compared with normal group (170.68 pg/ml ± 42.46 pg/ml) respectively (F = 14.319, P < 0.001). Significantly increased organ vasopermeability of liver, kidney, lung and heart was observed after CLP respectively. The number of CEC peaked (11.83 ± 1.94) 3 hours after CLP compared with normal control (5.33 ± 1.21, P < 0.05), and then decreased gradually (F = 54.183, P < 0.001). The mRNA level of Tie2 in CLP-3 h group (3.47 ± 1.47) was also markedly higher than that in other groups (F = 10.640, P < 0.001).
Using quantitative real-time PCR to measure the level of Tie2 mRNA in peripheral blood is a simple and relatively sensitive method to evaluate the damage of endothelial cells.
通过检测脓毒症大鼠模型中循环Tie2 mRNA水平来评估内皮细胞损伤情况。
采用盲肠结扎穿孔术（CLP）建立90只大鼠的脓毒症模型，将其分为6组：正常组、假手术组、CLP-3 h组、CLP-6 h组、CLP-12 h组和CLP-24 h组。用自动生化分析仪检测血清生化标志物。采用酶联免疫吸附测定法检测血清白细胞介素-6（IL-6）。用伊文思蓝检测肝、肾、肺和心脏的血管通透性。采用密度梯度分离法分离循环内皮细胞（CEC）并计数。提取全血总RNA，通过定量实时聚合酶链反应检测两种内皮特异性基因Tie2和血管内皮生长因子受体2（VEGFR2）的mRNA水平。
CLP术后血清生化指标水平升高。CLP-6 h组、12 h组和24 h组血清IL-6含量分别比正常组（170.68 pg/ml±42.46 pg/ml）升高6.5倍（P<0.05）、8.4倍（P<0.01）和13.3倍（P<0.001）（F=14.319，P<0.001）。CLP术后肝、肾、肺和心脏的器官血管通透性分别显著增加。与正常对照组（5.
