摘要
目的用蛋白酶和蛋白酶激活受体激动肽(PAR-AP)刺激A549细胞后检测细胞中白细胞介素IL-28和IL-29 mRNA表达情况以分析蛋白酶激活受体激活对人肺上皮细胞IL-28和IL-29基因表达的影响。方法用最佳工作浓度的凝血酶、胰蛋白酶、类胰蛋白酶及蛋白酶激活受体-1,2,3,4的激动肽刺激A549细胞后,分别在第2、8、16h收集细胞,用实时定量逆转录聚合酶链式反应(q-RT-PCR)方法分析A549细胞内IL-28和IL-29 mRNA表达。结果凝血酶作用A549后,细胞内IL-29 mRNA表达增强,高达对照组9.6倍;而细胞内IL-28 mRNA表达无明显变化。胰蛋白酶作用A549细胞后,细胞内IL-29和IL-28 mRNA表达增强,分别为对照组6.1倍和4.4倍。类胰蛋白酶作用A549细胞后,细胞内IL-29和IL-28 mRNA表达增强分别高达对照组的3.1倍和2.1倍。弹性蛋白酶作用A549细胞后,细胞内IL-29 mRNA表达增强高达对照组5.1倍而IL-28 mR-NA无明显变化。PAR-1AP(SFLLR)作用后,A549细胞内IL-29 mRNA表达增强为对照组1.7倍而IL-28 mRNA表达无明显变化。PAR-2AP(SLIGKV及tc-LIGRLO)作用后,A549细胞内IL-29和IL-28 mRNA表达增强分别为对照组4.9倍和2.4倍。PAR-3AP(TFRGAP)作用后,A549细胞内IL-29和IL-28 mRNA表达无明显变化。PAR-4AP(GYPGQV)作用后A549细胞内IL-29和IL-28 mRNA表达增强高达对照组4.1倍和5.5倍。结论 A549细胞蛋白酶激活受体的激活可以上调细胞内IL-29和IL-28 mRNA表达。
This study is aimed to detect the effect of protease-activated receptor activation on IL-28 and IL-29 mRNA expression in A549 cells. Firstly, A549 cells were collected at 2 h, 8 h and 16 h after stimulation with the optimal concentrations of thrombin, trypsin, tryptase and elastase and protease-activated receptors＇ activated pepfide （PARs AP）, then the cells were treated for detecting IL-28 and IL-29 mRNA expression by using quantitative real time polymerase chain reaction （PCR） technique. We found that thrombin upregulated IL-29 mRNA expression by up to 9.6-fold of control, but had little effect on regulation of IL-28 mRNA expression in A549 cells. In addition, trypsin enhanced expressions of IL-29 and IL-28 mRNA by up to 6.1- and 4.4-fold of control, respectively, while tryptase increased expressions of IL-29 and IL-28 mRNA by up to 3.1- and 2.2-fold of control respectively in A549 cells. Elastase stimulated IL-29 mRNA expression by up to 5.1-fold of control, but had little influence on the expression of IL-28 mRNA in A549 cell. The PAR-1 AP, SFLLR, induced upregulated expression of IL-29 mRNA by up to 1.7-fold of control whereas PAR-1 AP exerted little effect on IL-28 mRNA expression accordingly. Furthermore, PAR-2 AP （SLIGKV） or tc-LIGRLO induced up to 4.9- and 2.4-fold increase in IL-29 and IL-28 mRNA expression, while PAR-4 AP （GYPGQV） strengthened IL-29 and IL-28 mRNA expression by up to 4.1- and 5.5-fold of control, respectively. Whereas, PAR-3 AP （TFRGAP） had little effect on IL-29 and IL-28 mRNA expression. In conclusion, activation of PARs provoked up-regulation of IL-28 and IL-29 mRNA in A549 cells.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2010年第10期833-837,841,共6页
Immunological Journal
基金
国家自然科学基金(30772032
30972714
81060250)
海南省自然科学基金(808156)
海南省教育厅科研项目(Hj2009-129)
