摘要
目的探讨核因子-κB(NF-κB)在急性胰腺炎体外模型中的作用。方法以10 mg／L脂多糖刺激AR42J细胞构建急性胰腺炎的体外模型,设2 h、6 h、12 h、18 h和24 h共5个时间点,每时间点设3个复孔,逆转录-多聚酶链反应(RT-PCR)半定量法观察细胞间黏附分子-1(ICAM-1)和NF-κB p65亚单位mRNA表达的改变;链酶亲和素-生物素-过氧化物酶复合物法(SABC)检测p65蛋白在AR42J细胞中的表达;人工碘比色法观察培养液上清中淀粉酶的改变。结果脂多糖刺激后,AR42J细胞以时间依赖方式上调ICAM-1 mRNA和p65 mRNA的表达,24 h达最高值;二者之间具有直线相关性(P< 0．01),同时p65蛋白亦呈时间依赖方式表达增强,24 h表达最强;各组间淀粉酶无明显改变(P>0．05)。结论在脂多糖诱导的胰腺腺泡细胞AR42J炎性效应中,NF-κB调控致炎细胞因子ICAM-1的表达。
Objective To explore the effects of NF-kappa B（NF-κB） in vitro model of acute pancreatitis. Methods AR42J cell line was stimulated by lipopolysaccharide （LPS） with 10 mg/L for different hours （2 h, 6 h, 12 h, 18 h and 24 h）. We set up 3 holes at every point time. The expression changes of ICAM-1 and p65 mRNA level were detected by reverse transcription-polymerase chain reaction （RT-PCR）, and the expression of p65 protein in AR42J cell lines was detected by Stept Avidin- Biotin Complex, and the changes of amylase in culture medium by chromatometry. Results LPS could induce the expression of 1CAM-lmRNA and p65 mRNA in a manner of time dependency. Analysis of correlation and regression indicated that there was positive correlation between the expressions of ICAM-1 mRNA and p65mRNA （P 〈 0. 01）. The expression of p65 protein increased gradually in a manner of time dependency after induction, but the amylase didn＇t, change obviously （P 〉 0. 05）. Conclusions NF-κB controlled the expression of proinflammatory cytokine ICAM-1 in the lipopolysaccharide induced inflammatory effects on rat pancreatic acinus AR42J cell lines.
出处
《胰腺病学》
2006年第4期204-207,共4页
Chinese JOurnal of Pancreatology
基金
广东省自然科学基金自由申请重点项目(51342001)
