组织因子在新生鼠坏死性小肠结肠炎肠组织中的表达

The expression of tissue factor in intestines of necrotizing enterocolitis rats

目的 研究新生鼠坏死性小肠结肠炎（NEC）模型肠组织中组织因子（TF）的表达,探讨TF在NEC发生中的作用。方法 40只新生Wistar大鼠,出生36~48 h,随机分为正常对照组（n=10）和实验组（n=30）。实验组予鼠乳代用品人工喂养、给予100%氮气缺氧90 s,4℃冷刺激10 min（每天2次,连续2 d）。实验结束后处死动物。留取回盲部附近肠组织进行病理损伤评分和肠组织中TF含量检测。组织学评分≥2确定为NEC,运用RT-PCR、Western Blot等方法,研究NEC模型肠组织中TFm RNA、TF蛋白的表达。结果 肠组织TFm RNA表达水平（2^-ΔΔCt）：实验组为（3.27±1.28）,对照组为（1.01±0.04）,二者比较,差异有统计学意义（F=21.67,P〈0.01）;肠组织TF蛋白表达（灰度值）：实验组为（11.42±0.27）,对照组为（7.21±0.33）,二者比较,差异有统计学意义（H=17.15,P〈0.01）。实验组肠组织TFm RNA及蛋白的表达均增加。结论 从转录及翻译水平证实,TF可能参与NEC的发生发展。

Objective To study the expression of TF in the intestines of NEC model, and investigate the possible role of TF in the development of NEC. Methods 40 Wistar newborn rats （age 36~48 h） were randomly divided into control group including 10 rats and experimental group including 30 rats. The rats in experimental group were given formula feeding （and were exposed to hypoxia and cold exposure （hypoxia treatment for 90 seconds by 100%nitrogen, 4℃cold stimulation for 10 minutes,twice a day and lasted for 2 days）. Then the rats were sacrificed. Intestinal tissue samples located at the boundary of ileum and cecum were obtained for HE staining checking mRNA and protein of TF. mRNA and protein of TF in intestinal tissues were detected by Real-time quantitative reverse transcription polymerase chain reaction （QRT-PCR）and Western Blot methods. Results Levels of intestinal TF mRNA （2^-ΔΔCt value） were 3.27 ±1.28 （experiment group）,and 1.01 ±0.04 （control group）; levels of intestinal TF protein were 11.42±0.27 （experiment group） and 7.21±0.33 （control group）.There were significant differences of mRNA and protein levels of TF between the two groups （P〈0.01）. Conclusion The level of TF in intestines of NEC model in newborn rats was increased , which suggested that TF probably be involved in the process of NEC.